In-Vitro Diagnostic Devices: Introduction to

Current Point-of-Care Diagnostic Devices 1st

Edition Chao-Min Cheng pdf download

https://textbookfull.com/product/in-vitro-diagnostic-devices-
introduction-to-current-point-of-care-diagnostic-devices-1st-
edition-chao-min-cheng/

Download more ebook instantly today - get yours now at textbookfull.com

We believe these products will be a great fit for you. Click
the link to download now, or visit textbookfull.com
to discover even more!

Diagnostic devices with microfluidics 1st Edition

Francesco Piraino

https://textbookfull.com/product/diagnostic-devices-with-
microfluidics-1st-edition-francesco-piraino/

III V Compound Semiconductors and Devices An

Introduction to Fundamentals Keh Yung Cheng

https://textbookfull.com/product/iii-v-compound-semiconductors-
and-devices-an-introduction-to-fundamentals-keh-yung-cheng/

Electronic Devices Conventional Current Version 10th

Edition Thomas L. Floyd

https://textbookfull.com/product/electronic-devices-conventional-
current-version-10th-edition-thomas-l-floyd/

Energy Efficient Computing Electronics Devices to

Systems Devices Circuits and Systems 1st Edition
Santosh K. Kurinec

https://textbookfull.com/product/energy-efficient-computing-
electronics-devices-to-systems-devices-circuits-and-systems-1st-
edition-santosh-k-kurinec/
Designing for Wearables Effective UX for Current and
Future Devices 1st Edition Scott Sullivan

https://textbookfull.com/product/designing-for-wearables-
effective-ux-for-current-and-future-devices-1st-edition-scott-
sullivan/

Artificial Intelligence in Cancer: Diagnostic to

Tailored Treatment 1st Edition Smaranda Belciug

https://textbookfull.com/product/artificial-intelligence-in-
cancer-diagnostic-to-tailored-treatment-1st-edition-smaranda-
belciug/

Primer of diagnostic imaging Sixth Edition Chen

https://textbookfull.com/product/primer-of-diagnostic-imaging-
sixth-edition-chen/

Diagnostic Imaging in Polytrauma Patients 1st Edition

Vittorio Miele

https://textbookfull.com/product/diagnostic-imaging-in-
polytrauma-patients-1st-edition-vittorio-miele/

Pocket Guide to Diagnostic Hematopathology S. David

Hudnall

https://textbookfull.com/product/pocket-guide-to-diagnostic-
hematopathology-s-david-hudnall/
Chao-Min Cheng · Chen-Meng Kuan
Chien-Fu Chen

In-Vitro
Diagnostic
Devices
Introduction to Current Point-of-Care
Diagnostic Devices
In-Vitro Diagnostic Devices
Chao-Min Cheng · Chen-Meng Kuan
Chien-Fu Chen

In-Vitro Diagnostic Devices

Introduction to Current Point-of-Care
Diagnostic Devices

13
Chao-Min Cheng Chien-Fu Chen
National Tsing Hua University National Chung Hsing University
Hsinchu Taichung
Taiwan Taiwan

Chen-Meng Kuan
National Tsing Hua University
Hsinchu
Taiwan

ISBN 978-3-319-19736-4 ISBN 978-3-319-19737-1 (eBook)

DOI 10.1007/978-3-319-19737-1

Library of Congress Control Number: 2015941344

Springer Cham Heidelberg New York Dordrecht London

© Springer International Publishing Switzerland 2016
This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part
of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations,
recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission
or information storage and retrieval, electronic adaptation, computer software, or by similar or
dissimilar methodology now known or hereafter developed.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this
publication does not imply, even in the absence of a specific statement, that such names are exempt
from the relevant protective laws and regulations and therefore free for general use.
The publisher, the authors and the editors are safe to assume that the advice and information in this
book are believed to be true and accurate at the date of publication. Neither the publisher nor the
authors or the editors give a warranty, express or implied, with respect to the material contained
herein or for any errors or omissions that may have been made.

Printed on acid-free paper

Springer International Publishing AG Switzerland is part of Springer Science+Business Media

(www.springer.com)
Contents

1 Introduction to In Vitro Diagnostic Devices. . . . . . . . . . . . . . . . . . . . . . 1

1.1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
1.2 Structure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
1.3 Advantages. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.4 Antibody. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.5 Labels. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
1.6 Membranes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.7 Application. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
1.8 Conclusion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

2 Polymeric-Based In Vitro Diagnostic Devices. . . . . . . . . . . . . . . . . . . . . 15

2.1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.2 Selection of Polymer Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.2.1 Polydimethylsiloxane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.2.2 Cyclic Olefin Copolymer. . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.3 Fabrication of Polymer Devices. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.3.1 Structure Formation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.3.2 Device Sealing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
2.3.3 World-to-Chip Interface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
2.4 Fluidic Control Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
2.4.1 Valve. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
2.4.2 Pump. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
2.4.3 Mixer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
2.5 Applications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
2.5.1 Sample Preparation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
2.5.2 Separation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
2.5.3 Reagent Storage. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
2.5.4 Detection of Metabolites and Small Molecules. . . . . . . . . . . 42

v
vi Contents

2.5.5 DNA- and RNA-Based Diagnosis. . . . . . . . . . . . . . . . . . . . . 43

2.5.6 Protein-Based Diagnosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
2.5.7 Cell Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

3 Low-cost In Vitro Diagnostic Technologies. . . . . . . . . . . . . . . . . . . . . . . 59

3.1 Overview of Low-cost In Vitro Diagnostic Technologies . . . . . . . . . 59
3.2 Paper-Based Microfluidic Devices. . . . . . . . . . . . . . . . . . . . . . . . . . . 60
3.2.1 Benefits of Paper . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
3.2.2 Fabrication Techniques. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
3.2.3 Detection Methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
3.2.4 New Functions and Design. . . . . . . . . . . . . . . . . . . . . . . . . . . 71
3.2.5 Diagnostic Applications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
3.3 Thread-/Cotton-Based Microfluidics. . . . . . . . . . . . . . . . . . . . . . . . . 85
3.4 Commercialization of Low-cost Microfluidic
Devices for Clinical Diagnostics. . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
3.5 Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88

4 Glucose Sensor and Its Potential Directions. . . . . . . . . . . . . . . . . . . . . . 93

4.1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
4.2 Design and Fabrication of the Contact
Lens-Based Glucose Sensor. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
4.2.1 Glucose Sensor Design and Fabrication. . . . . . . . . . . . . . . . . 99
4.2.2 LED (Red Light) Fabrication. . . . . . . . . . . . . . . . . . . . . . . . . 100
4.2.3 Antenna Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100
4.2.4 Wireless Readout Chip Architecture . . . . . . . . . . . . . . . . . . . 101
4.2.5 Fabrication for the Integration of Radio
and Sensor with Contact Lens . . . . . . . . . . . . . . . . . . . . . . . . 102
4.3 Potential Directions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104
Chapter 1
Introduction to In Vitro Diagnostic Devices

1.1 Overview

Healthcare investment keeps on increasing substantially in recent years [1, 2].

Such investment has also focused on fighting major diseases, enabled by the novel
invention of cost-effective and valid drug development for treatment and side effect
reduction, along with improved vector control. In addition, the demand for diag-
nostics that is essential in determining prognosis, identifying disease stages, moni-
toring treatment, and assessing the spreading as health services has expanded [3].
Molecular-based diagnostics is critical for prevention, identification, and treat-
ment of disease. Current laboratory analyses support correct diagnosis in over
70 % of all diseases and can be used to aid the continuous monitoring of drug
therapy [4]. Traditional diagnosis system in central laboratory is therefore a vital
component in the clinic and in local general practice. However, classic diagnos-
tic technologies are not completely well suited to meeting the expanded testing
requirement because they rely on complicated sample purification and sophisti-
cated instruments which are labor-intensive, timely, and expensive and require of
well-trained operators. One of the main challenges for industry is to develop fast,
relatively accurate, easy-to-use, and inexpensive devices. For example, micros-
copy observation requires less infrastructure and is more widely available based
on the simplicity and low cost; however, the accuracy is somehow questionable
and underutilized (e.g., smear tests for tuberculosis, malaria, and schistosomiasis)
[5–7]. As a result, it not only increases the cost and inconvenience of health care
but also causes patients to leave the medical system before the diagnostic result
is obtained [8]. Faster and more accurate diagnostic tests that require minimum
laboratory equipment and operation training play an important role in expanding
health care in resource-constrained settings [9, 10].
In addition to the improved efficiency in laboratory diagnostics, there has been
a trend toward a more decentralized diagnostics which occurs directly at patients’

© Springer International Publishing Switzerland 2016 1

C.-M. Cheng et al., In-Vitro Diagnostic Devices, DOI 10.1007/978-3-319-19737-1_1
2 1 Introduction to In Vitro Diagnostic Devices

bedside, in outpatient clinics, or at the sites of accidents, so-called point-of-care

(POC) systems [11]. The concept of POC testing is mainly for the patient, so short
turnaround time, minimum sample preparation and reagent storage and transfer-
ring, user-friendly analytical instruments, and digital or visible quantitative or
semiquantitative single readout is required [4, 12, 13]. It is clear that on-site or
minimum sample preparation and on-chip storage limit the delays that caused by
transport and preparation of clinical samples. As a result, shorter turnaround time
leads to rapid clinical decision-making and may save fatal consequences. No pre-
vious knowledge in sample analysis should be required, so elders can perform the
tests at home with minimum training to improve health outcome [14].
The first POC device was urine dipstick test, which was developed in 1957 to
measure urinary protein [15]. Glucose meters for diabetic monitoring and lateral-
flow devices for pregnancy tests are currently the most widely used devices in
POC molecular diagnostics. They are excellent examples of POC tests; however,
they are still not applicable if highly sensitive and high-throughput quantitative
measurements are required.
In recent decades, some technologies have emerged that fulfill these require-
ments. Lateral-flow immunoassay (LFIA) devices, for example, which were
originally proposed in the 1980s, remain popular largely because of their design
simplicity.
Plotz and Singer invented the latex agglutination assay in 1956, from which
the technical basis for the LFIA was derived [16]. Plate-based immunoassay was
being developed at the same time. The radioimmunoassay was designed by Berson
and Yalow in the 1950s [17]. The enzyme immunoassay, which replaced radioi-
sotopes with enzymes, cut down reaction times, and provided higher specificities
than a radioimmunoassay, was developed in the 1960s. The fundamental principles
of the LFIA continued to be refined through the 1980s and were firmly established
during the ensuing years [18, 19]. Since that time, at least another 500 patents
have been filed on various aspects of the technology. Several patents have even
been formatted by companies such as Becton Dickinson & Co. and Unilever and
Carter Wallace.
The chief application driving the early development of solid-phase, rapid-test
technology was the human pregnancy test, which was symbolic of continued his-
torical interest in urine testing for medical diagnostic purposes. This particular
testing application made great strides in the 1970s, as a result of improvements
in antibody generation technologies and significant gains in understanding the
biology and detection of human chorionic gonadotropin (hCG), derived largely
from the work performed by Vaitukaitis and colleagues [20]. However, to entirely
evolve the lateral-flow test, considerable enabling technologies were still required.
Many of these technologies, such as nitrocellulose membrane manufacturing,
antibody generation, and processing equipment, were developed throughout the
1990s. The purpose of this article is to introduce readers with basic information
regarding the LFIA approach.
1.2 Structure 3

1.2 Structure

Figure 1.1 displays the key elements of a LFIA. This assay consists of several
components, often segmented parts made of different materials. When a test is
run, appropriately conditioned sample is added to the proximal end of the strip, the
absorbent pad. The treated sample then migrates to the conjugate pad, where an
appropriate reagent has been immobilized. The labeled reagent on the conjugate
pad can be colloidal gold, or a colored, fluorescent, or paramagnetic latex particle.
These specific biological components can be either antigen or antibody depending
on the assay format. Next, the sample remobilizes the dried reagent, and particle
interaction ensues. Sample and reagent then migrate to the next segment of the
strip, the reaction matrix. The reaction matrix is a porous membrane, upon which a
final specific biological component has been immobilized. These biological com-
ponents are usually proteins, either antibody or antigen. They have been bound
onto the specific lines of the membrane being used. As the sample and reagent
reach this line, they are captured by the applied proteins, and excess liquid moves
past this point and is taken up by the absorbent pad. The result is the detectable
absence or presence of the test line, read by eyes or by other instruments.
The LFIA may be of two different types: (1) direct (sandwich, Fig. 1.2a) or (2)
competitive (inhibition, Fig. 1.2b). Both types can accommodate qualitative, semi-
quantitative, and fully quantitative determinations. Direct assay is usually used
when testing for larger analytes with multiple antigenic sites, such as hCG, dengue
antigen, or human immunodeficiency virus (HIV). A positive result is indicated by
the presence of a test line. The conjugated particles also reach and are captured at
the control line. The control line typically comprises a species–specific anti-immu-
noglobulin antibody, specific for the antibody in the conjugate pad. Competitive
assay is usually used when testing for small molecules with single antigenic deter-
minants that cannot bind to antibodies on a test line simultaneously. In such cases,
a positive result is indicated by the absence of a test line, but a control line may
still form.

Sample

Absorbent Conjugate pad Membrane Test line Control line Plastic Absorbent
pad with reagent backing pad

Fig. 1.1  Typical structure of a LFIA strip

4 1 Introduction to In Vitro Diagnostic Devices

Test
(a) Conjugate
line
Control
pad line
Sample /
Analyte
Pre-run strip

Positive result

Negative result

(b) Conjugate Test Control

pad line line

Sample /
Analyte Pre-run strip

Negative result

Positive result

Fig. 1.2  a Direct solid-phase immunoassay. b Competitive solid-phase immunoassay

1.3 Advantages 5

1.3 Advantages

LFIAs represent a well-established and very appropriate technology when applied

to a wide variety of in vitro diagnostics (IVD) or field-use applications. The
advantages of the LFIA are well known:
a. Technology is mature.
b. Manufacture is relatively easy: Equipment and processes are already developed
and available.
c. They can be scalable to high-volume production.
d. They can be stored for 12–24 months, often without refrigeration.
e. They are easy to use, requiring minimal operator-dependent steps and
interpretation.
f. They can handle small volumes of multiple sample types.
g. They can be integrated with onboard electronics, reader systems, and informa-
tion systems.
h. They have high sensitivity, specificity, and good stability.
i. Development and approval are relatively low cost and require a short timeline.
j. They are already present and accepted by the market: Minimal education is
required for users and regulators.

1.4 Antibody

Although the physical components of the lateral-flow test strip and construction
techniques play a major role, the most critical part of the LFIA is the appropri-
ate antibody to provide antigen recognition. If we chose inappropriate antibody, it
would not have ability to recognize the target antigen. Much time is spent deter-
mining the most suitable antibody for specific assays. Many scientists have spent a
great deal of time figuring out the suitable antibodies to fit the assay.
The LFIA is particularly demanding in terms of the mass of the reagent used
to drive the antibody and antigen interaction. When an antibody is used in a sand-
wich-type assay, they are applied at a ratio of 1–3 µg per cm across the width of
the nitrocellulose strip, in a line 1 mm wide and with a relatively shallow bed vol-
ume of 0.13 mm. This results in an antibody concentration of 10–30 µg per square
cm, which is 25–100 times that used in an enzyme-linked immunosorbent assay
(ELISA), which can typically require a maximum concentration of 300 ng per
square cm [21].
Antibody and antigen affinity also plays an important role in the assay.
Consider a typical lateral-flow test strip with antibody immobilized on a test line
of 0.5–1.0 mm wide. Antigen flowing up the strip has a flow rate in the range of
0.16–0.66 mm per second, depending on the flow rate of the nitrocellulose mem-
brane selected [22]. Antigen thus spends between 1–6 s on the line where it
can interact with the immobilized antibody. Flow speed is actually faster at the
6 1 Introduction to In Vitro Diagnostic Devices

initiation of the flow, since the flow rates decrease proportionately to the square of
the distance traveled, a steady flow rate is achieved, and the entire nitrocellulose
bed volume becomes saturated.
Antibodies applicable for LFIA are available from many commercial sources
[19]. Frequently, these antibodies can be obtained for competitive assay, such as
hormones, therapeutic drugs, and drugs of abuse. Similarly, suitable antibodies are
purchasable for sandwich assay tests to diagnose pregnancy (hCG), infectious dis-
ease (HIV, hepatitis B), cardiac markers (troponin C, creatinine kinase-MB, myo-
globin), or malignancies (prostate-specific antigen).

1.5 Labels

Some labels have been successfully commercialized and others appear promising.
The development of labels for LFIA has matured hand-in-hand with advances in
detection methodology and instrumentation. Sensitive assays with fluorescent and
luminescent labels have been used in recent years. The ideal labels for lateral-flow
strips have the following characteristics:
a. They can be detected by multiple methods on a large and useful dynamic range.
b. When sample and reagent conjugate, their biological and chemical quality and
activity are not be changed.
c. The lack of non-specific binding characteristic such as high signal-to-noise
ratio under buffer, salt, or detergent conditions.
d. High stability under various temperatures.
e. They are typically available at low cost.
f. The procedure of conjugating is easy and scalable.
g. They are capable of being used for multianalyte detection.
Liposomes can be used as a vehicle for membrane-based assays in vertical and
lateral-flow test strips (e.g., test for malarial antigen from Becton Dickinson) [20].
Because of their ability to encapsulate very high concentrations of signal-generat-
ing molecules within their cores, liposomes can improve LFIA sensitivity to 2–3
orders. Lipoproteins, glycolipids, and various other lipid-containing compounds
can be incorporated directly into the bilayer. In addition, different chemically
active groups can be incorporated onto the lipid surface with controlled surface
density for covalent coupling to biological or chemical compounds [23].
Colloidal carbon particles can serve as a label in sol particle immunoassays
[24]. They have been reported since the 1970s [25]. Their advantages include good
stability and high color comparison on a membrane. They are quite easy to conju-
gate, and a bottle of carbon particles may consequently last for millions of tests.
Colloidal gold has been widely used in immunoassays for large molecules
such as for the detection of hormones (pregnancy, fertility), virus (HIV, hepatitis
B and C), and bacteria (Streptococcus suis serotype 2). It may be the most widely
used label today [26]. Determination via colloidal gold-based immunoassay can
1.5 Labels 7

be completed rapidly in a single step [27]. When an antibody labeled with colloi-
dal gold particles is combined with the corresponding antigen, the colored immu-
noreactant can be visually detected. This user-friendly format possesses several
advantages, including rapid reaction time, long-term stability over a wide range of
climates, and low cost. These characteristics make it ideally suited for on-site test-
ing by untrained personnel.
A variety of other labels have been used for specific applications. For instance,
a portable fluorescence biosensor with rapid and ultrasensitive response for protein
biomarker has been created using quantum dots and a LFIA. The superior signal
brightness and high photostability of quantum dots are combined with the prom-
ising advantages of a lateral-flow test strip, resulting in high sensitivity, selectiv-
ity, and speed for protein detection [28]. Also, more recent reporter up-converting
phosphor technology has been applied to DNA (hybridization) assays for the
detection of specific nucleic acid sequences. This methodology is sensitive and
provides a rapid alternative for more elaborate gel electrophoresis and Southern
blotting [29].

1.6 Membranes

While a LFIA test strip may include elegant chemical complexity, the common
core of all such tests is the nitrocellulose membrane, which for several reasons is
the most significant test component [30–32]. First, it is the surface upon which the
critical immune complexes form. Second, it is the surface upon which the signal
is detected, either visually or electronically. Third, it has been the most difficult
material to manufacture consistently.
One of the key membrane performance parameters is protein binding. It is
essential to the function of the membrane in a lateral-flow test strip. The mem-
brane usually adsorbs more than 100 μg of IgG per cm2. At the concentrations
of capture reagents typically applied to the membrane, there is fivefold to ten-
fold more binding capacity than necessary. Adsorptive capacity decreases with
the molecular weight of the protein [33]. To maximize adsorption, antibodies and
other proteins should be applied to the membrane in buffers that are preferably
free of salt, surfactants, and sugars. The buffer should also be at a low concen-
tration so that crystals dried in the membrane are not of sufficient abundance to
occlude the pores.
Another key membrane performance parameter is membrane blocking.
Blocking prevents non-specific binding of the detector particle and analyte, but is
not absolutely essential to LFIA strips. There are many test strips on the market
that do not use a blocking agent; however, blocking agents are required for some
tests because of the nature of the particular sample and antibody system [34]. Two
blocking agents must be used: one blocking agents dissolves upon addition of the
sample and moves along the strip with sample, and the other is applied directly to
the membrane by spraying on a fixed amount of blocking solution or dipping the
membrane into a reservoir of blocking solution.
8 1 Introduction to In Vitro Diagnostic Devices

The final membrane performance parameter to consider is membrane storage

capacity. Storage capacity and condition vary depending on the stage of the test
strip manufacturing process. Up until the point that reagent is going to be applied,
the membrane can be stored under ambient conditions (15–30 °C, 20–80 % relative
humidity). A condensing atmosphere should be avoided, as liquid in the pores can
cause redistribution of mobile components, such as the surfactant. When a mem-
brane is being prepared for application of the capture reagents, it should be allowed
to equilibrate to the humidity of the dispensing room. Humidity from the air hydrates
the surface of the nitrocellulose and improves the absorption of the capture reagent
solutions. If possible, assembly of the test strips should take place in a dry room.

1.7 Application

LFIA is well established as a valuable tool in food, medical, environmental, vet-

erinary, agricultural, and industrial diagnostics. Sometimes it is used as a rapid
screening tool and backed up by more complex and time-consuming assays.
Figure 1.3 lists the market segments in which LFIAs are already in production or
are known to be in development.

Medical Food
Diagnostic Safety
Industrial*
Blood
Blanking

Therapeutic
Animal
Monitoring
Health
Application
Consumer
Environmental
Diagnostic

Military/
Aquaculture
Biodefense
*includes applications such as
Agriculture
Forensic QC, product identification,
Science environmental monitoring,

safety

Fig. 1.3  Market segments for LFIA and other point-of-care or field-use technologies
1.7 Application 9

LFIA has well-established formats for POC testing. The first paper-based
diabetes dipstick test was created in the 1950s to quantify glucose in urine [35].
Semiquantitative results could be determined by comparing urine-treated test
strips to a color-coded chart to determine glucose concentration. Today, commer-
cial urinalysis dipsticks have been widely adapted for a number of analyses. In
the 1980s, serological lateral-flow tests started to emerge, particularly for human
pregnancy tests. This process was derived from the development of the hCG beta-
subunit radioimmunoassay [36, 37].
The majority of these tests come in different sizes, shapes, and configurations.
These assays are available without (Fig. 1.4a, b) or with housing units (Fig. 1.4c–f).
Nowadays, multiplexing of rapid tests is becoming fairly common as illustrated in
Fig. 1.4g–i, which illustrates a lateral-flow format that separates each single lateral-
flow test strip into multiple channels. The assay is multiplexed in the sense that a
single sample is analyzed simultaneously, but in reality, the test strips are still sepa-
rate reactions occurring independently of the other reactions [38, 39].
On-chip reagent storage for long-term test and transportation is well devel-
oped for IVD. For example, LFIA strips adopt dried gold nanoparticles (AuNPs)-
conjugated antibodies regents at conjugation pad for rapid pregnancy, drug abuse,
and other diagnostic tests. A plasma fibrinogen assay was implemented on a poly-
meric micropillar-based LFIA platform by drop-casting bovine thrombin and the
surfactant Triton X-100 on the dextran-coated platform [40]. This pillar structure
can also be used for an interferon-γ LFIA assay [41].
One of the major application for IVD test is the detection of the metabolites of
illegal drugs such as Δ9-tetrahydrocannabinol (THC), amphetamines, benzodiaz-
epines, cocaine, morphine, heroin, opiates, and cannabis in workplace or prison
settings. The presence of addictive drugs in the body fluids including blood, urine,
sweat, and saliva is monitored to detect and prevent drug abuse, illicit trafficking
or driving under the influence of drug (DUID) that is getting more attention world-
wide [42, 43]. Furthermore, continuous concern about recreational drug abuse and
doping in competitive sports still attracts social attention [44, 45]. The prohibited
substances such as strychnine, pervitin, captagon, or Benzedrine are the target
molecules for detection.
Oral fluid has been demonstrated as an adequate alternative matrix for drugs
identifying and quantifying tests in workplace, clinical treatment, drug rehabili-
tation center, criminal justice, and DUID settings [46]. The drug tests using oral
fluid instead of blood and urine possess various advantages such as inexpensive,
rapid, infection risk is lower than for blood sample, and noninvasive of sample
collection, which can be easily observed to avoid the need for private facilities and
same-sex collectors and decrease adulteration. In addition, oral fluid better reflects
recent drug use and reflects free plasma concentrations, providing a better correla-
tion with pharmacodynamic effects.
Liquid chromatography–tandem mass spectrometry (LC-MS/MS) and gas
chromatography–tandem mass spectrometry (GC-MS/MS) are the most delegated
equipment performing high accurate analysis of multiple compounds in a limited
oral fluid volume. However, the complex sample preparation using liquid–liquid
10 1 Introduction to In Vitro Diagnostic Devices

(a) (b)

(c) (d)

(e) (f)

(g) (h) (i)

Fig. 1.4  Commercial LFIA tests. a Determine™ HIV 1/2 Ag/Ab Combo. © 2013 Alere. All rights
reserved. b Determine™ TB-LAM Ag test © 2013 Alere. All rights reserved. c One Step LH Ovu-
lation Rapid Test © 2010 Accu Plus Medical. All rights reserved. d Clearview® Malaria P.f. Test ©
2013 Alere. All rights reserved. e ICON HP © Beckman Coulter, Inc. All rights reserved. f BD™
EZ Flu A + B Test © Becton Dickinson. g RAID™ 5 © Alexeter Technologies. h SNAPduo™
Beta-Tetra ST Test © 2013 IDEXX Laboratories, Inc. (https://www.idexx.com/small-animal-health/
index.html; accessed 10/15/2014). i SNAP® Heartworm RT Test © 2012 IDEXX Laboratories, Inc.
(https://www.idexx.com/small-animal-health/index.html; accessed 10/15/2014)
1.7 Application 11

extraction or solid-state extraction, time-consuming detection processes, bulky

size of equipment, and power sources requirement confined the possibility of
on-site tests. There are some commercial portable oral fluid test devices that have
been developed and available on market providing satisfactory detection ability to
achieve the requirement of detection limit of certain drugs. One of the successful
commercial examples for on-site drug test is Oratect. It is a LFIA-based test utiliz-
ing AuNPs for colorimetric sensing. In order to collect oral fluidic samples, sam-
ple collector is combined in a single device [47].
A number of strategies are available for the detection of nucleic acids in lat-
eral-flow systems [48–50]. The capture of nucleic acids can be performed in
an antibody-dependent or antibody-independent way. For example, in an anti-
body-dependent system, an anti-biotin antibody immobilized on the surface of
nitrocellulose is used to capture biotin- and carboxyfluorescein (FAM)-bearing
oligonucleotides in RPA amplicons [51]. Binding is subsequently detected using
an anti-FAM-colloidal gold conjugate. An antibody-independent alternative uti-
lizes streptavidin as the binding agent. Immobilization of oligonucleotide probes
directly onto membranes is also possible using oligonucleotides linked to carrier
proteins.
When considering the worldwide market applicability of diagnostics, a socio-
economic division is often applied. Cardiac and other chronic diseases in the
expanding middle classes of emerging economies are growing, as are the inci-
dences of previously geographically limited infectious diseases (e.g., malaria,
dengue), emerging diseases (e.g., H5N1 influenza), and heretofore well-controlled
diseases (e.g., TB in First World countries) in developed countries. At least 30
previously unknown disease agents have been identified since 1973, including
HIV, Ebola, hepatitis C, and SARS. In chronic diseases, there remains significant
growth, particularly in the areas of inflammation, cardiac markers, and cancer,
with a myriad of new labels in development in the search for improved diagnostic
and prognostic indicators.
In the past 3–5 years, food safety issues and concerns for public health have led
to more stringent legislation in food safety requirements. Legislation has produced
increased demand for pathogen and toxin tests in just about every segment of the
food production industry. There is a growing demand from food companies for
quicker testing to facilitate more rapid release of finished goods and thus reduce
inventories. A driver in the demand for rapid and LFIA tests in food production
is the adoption of hazard analysis and critical control point (HAACP) regulations
that prescribe test procedures throughout the manufacturing process.

1.8 Conclusion

LFIA technology is rapidly being developed. Market needs lead to the improve-
ments in performance and utility and open doors to a vast array of new applica-
tion areas. With the integration of new reading, labeling, sample handling, and
12 1 Introduction to In Vitro Diagnostic Devices

device designs comes a requirement for a new approach to system development

and manufacturing. The development of highly sensitive and reproducible/quan-
titative next-generation point-of-need diagnostic assays requires a different, more
multidisciplinary approach than has been the case with standard LFIAs.

References

1. McCoy D, Chand S, Sridhar D (2009) Global health funding: how much, where it comes
from and where it goes. Health Policy Plann 24(6):407–417
2. Peeling R, Mabey D (2010) Point-of-care tests for diagnosing infections in the developing
world. Clin Microbiol Infect 16(8):1062–1069
3. Nkengasong JN, Nsubuga P, Nwanyanwu O, Gershy-Damet GM, Roscigno G, Bulterys M,
Schoub B, DeCock KM, Birx D (2010) Laboratory systems and services are critical in global
health time to end the neglect? Am J Clin Pathol 134(3):368–373
4. Luppa PB, Müller C, Schlichtiger A, Schlebusch H (2011) Point-of-care testing (POCT): cur-
rent techniques and future perspectives. TrAC Trends Anal Chem 30(6):887–898
5. Gray DJ, Ross AG, Li YS, McManus DP (2011) Diagnosis and management of schistosomia-
sis. BMJ: Br Med J 342
6. Lawn SD, Mwaba P, Bates M, Piatek A, Alexander H, Marais BJ, Cuevas LE, McHugh TD,
Zijenah L, Kapata N (2013) Advances in tuberculosis diagnostics: the Xpert MTB/RIF assay
and future prospects for a point-of-care test. Lancet Infect Dis 13(4):349–361
7. McNerney R, Daley P (2011) Towards a point-of-care test for active tuberculosis: obstacles
and opportunities. Nature Rev Microbiol 9(3):204–213
8. Rosen S, Fox MP (2011) Retention in HIV care between testing and treatment in sub-Saha-
ran Africa: a systematic review. PLoS Med 8(7):e1001056
9. Urdea M, Penny LA, Olmsted SS, Giovanni MY, Kaspar P, Shepherd A, Wilson P, Dahl CA,
Buchsbaum S, Moeller G (2006) Requirements for high impact diagnostics in the developing
world. Nature 444:73–79
10. Getahun H, Harrington M, O’Brien R, Nunn P (2007) Diagnosis of smear-negative pulmo-
nary tuberculosis in people with HIV infection or AIDS in resource-constrained settings:
informing urgent policy changes. The Lancet 369(9578):2042–2049
11. Jani IV, Peter TF (2013) How point-of-care testing could drive innovation in global health.
The New Engl J Med 368(24):2319–2324
12. Yager P, Domingo GJ, Gerdes J (2008) Point-of-care diagnostics for global health. Annu Rev
Biomed Eng 10:107–144
13. Price C, St John A, Hicks J (2004) Point-ofcare testing. American Association for Clinical
Chemistry, Washington DC
14. Price CP, Kricka LJ (2007) Improving healthcare accessibility through point-of-care technol-
ogies. Clin Chem 53(9):1665–1675
15. Unold D, Nichols JH (2010) Point-of-care testing: needs, opportunity, and innovation, by
Christopher P. Price, Andrew St John, and Larry J. Kricka, eds. Clin Chem 56(12):1893–1894
16. Plotz CM, Singer JM (1956) The latex fixation test: I. application to the serologic diagnosis
of rheumatoid arthritis. Am J Med 21(6):893–896
17. Berson SA, Yalow RS (1959) Quantitative aspects of the reaction between insulin and insu-
lin-binding antibody. J Clin Invest 38(11):1996–2016
18. Campbell RL, Wagner DB, O’Connell JP (1987) Solid phase assay with visual readout.
4703017 A, 1987-10-27
19. Rosenstein RW, Bloomster TG (1989) Solid phase assay employing capillary flow. 4855240
A, 1989-08-08
20. Moody A (2002) Rapid diagnostic tests for malaria parasites. Clin Microbiol Rev
15(1):66–78
References 13

21. Rowell V (2001) Nunc guide to solid phase. Roskilde, Nunc A/S
22. Rapid Lateral Flow Test Strips (2001) Considerations for product development. Millipore
Corporation, Bedford
23. Edwards KA, Baeumner AJ (2006) Analysis of liposomes. Talanta 68(5):1432–1441
24. van Amerongen A, Wichers JH, Berendsen LBJM, Timmermans AJM, Keizer GD, van
Doorn AWJ, Bantjes A, van Gelder WMJ (1993) Colloidal carbon particles as a new label
for rapid immunochemical test methods—quantitative computer image-analysis of results. J
Biotechnol 30(2):185–195
25. Geck P (1971) India-ink immuno-reaction for rapid detection of enteric pathogens. Acta
Microbiol Hung 18(3):191–196
26. Chandler J, Gurmin T, Robinson N (2000) The place of gold in rapid tests. IVD Technol
6:37–49
27. Wang S, Zhang C, Wang J, Zhang Y (2005) Development of colloidal gold-based flow-
through and lateral-flow immunoassays for the rapid detection of the insecticide carbaryl.
Anal Chim Acta 546(2):161–166
28. Li ZH, Wang Y, Wang J, Tang ZW, Pounds JG, Lin YH (2010) Rapid and sensitive detection
of protein biomarker using a portable fluorescence biosensor based on quantum dots and a
lateral flow test strip. Anal Chem 82(16):7008–7014
29. Corstjens P, Zuiderwijk M, Brink A, Li S, Feindt H, Neidbala RS, Tanke H (2001) Use of up-
converting phosphor reporters in lateral-flow assays to detect specific nucleic acid sequences:
a rapid, sensitive DNA test to identify human papillomavirus type 16 infection. Clin Chem
47(10):1885–1893
30. Jones KD (1999) Troubleshooting protein binding in nitrocellulose membranes. IVD Technol
5(2):32–41
31. Rapid Lateral Flow Test Strips (2002) Considerations for product development. Millipore
Corporation, Bedford
32. Beer HH, Jallerat E, Pflanz K, Klewitz TM (2002) Qualification of cellulos nitrate mem-
branes for lateral-flow assays. IVD Technol 8(1):35–42
33. Mansfield MA (2005) The use of nitrocellulose membranes in lateral-flow assays. In:
Wong RC, Tse HY (eds) Forensic science and medicine: drugs of abuse: body fluid testing.
Humana Press, Totowa, pp 71–85
34. Weiss A (1999) Concurrent engineering for lateral-flow diagnostics. IVD Technol 5(7):48–57
35. Free AH, Adams EC, Kercher ML, Free HM, Cook MH (1957) Simple specific test for urine
glucose. Clin Chem 3(3):163–168
36. Hawkes R, Niday E, Gordon J (1982) A dot-immunobinding assay for monoclonal and other
antibodies. Anal Biochem 119(1):142–147
37. Vaitukaitis JL, Braunstein GD, Ross GT (1972) A radioimmunoassay which specifically
measures human chorionic gonadotropin in the presence of human luteinizing hormone. Am
J Obstet Gynecol 113(6):751–758
38. Eldridge J (2000) Jane’s nuclear, biological and chemical Defence 2000–2001. Jane’s
Information Group Limited
39. Yetisen AK, Akram MS, Lowe CR (2013) Paper-based microfluidic point-of-care diagnostic
devices. Lab Chip 13(12):2210–2251
40. Dudek MM, Lindahl TL, Killard AJ (2010) Development of a point of care lateral flow
device for measuring human plasma fibrinogen. Anal Chem 82(5):2029–2035
41. Li JJ, Ouellette AL, Giovangrandi L, Cooper DE, Ricco AJ, Kovacs GT (2008) Optical scan-
ner for immunoassays with up-converting phosphorescent labels. IEEE Trans Biomed Eng
55(5):1560–1571
42. Gubala V, Harris LF, Ricco AJ, Tan MX, Williams DE (2011) Point of care diagnostics: sta-
tus and future. Anal Chem 84(2):487–515
43. Vearrier D, Curtis JA, Greenberg MI (2010) Biological testing for drugs of abuse. Molecular,
clinical and environmental toxicology. Springer, Berlin, pp 489–517
44. Deventer K, Roels K, Delbeke F, Van Eenoo P (2011) Prevalence of legal and illegal stimu-
lating agents in sports. Anal Bioanal Chem 401(2):421–432
14 1 Introduction to In Vitro Diagnostic Devices

45. Jelkmann W, Lundby C (2011) Blood doping and its detection. Blood 118(9):2395–2404
46. Gjerde H, Normann PT, Christophersen AS (2010) The prevalence of alcohol and drugs in
sampled oral fluid is related to sample volume. J Anal Toxicol 34(7):416–419
47. Wong RC, Tran M, Tung JK (2005) Oral fluid drug tests: effects of adulterants and food-
stuffs. Forensic Sci Int 150(2):175–180
48. Seal J, Braven H, Wallace P (2006) Point-of-care nucleic acid lateral-flow tests. IVD Technol
41
49. Dineva MA, Candotti D, Fletcher-Brown F, Allain JP, Lee H (2005) Simultaneous visual
detection of multiple viral amplicons by dipstick assay. J Clin Microbiol 43(8):4015–4021
50. O’Farrell B (2007) Sensitive, specific and rapid nucleic acid detection at the point of need
using simple, membrane-based assays. Bio World Eur 36–39
51. Piepenburg O, Williams CH, Stemple DL, Armes NA (2006) DNA detection using recombi-
nation proteins. PLoS Biol 4(7):e204
Chapter 2
Polymeric-Based In Vitro Diagnostic
Devices

2.1 Overview

The concept of translational medicine has begun to change biotechnology as it has

encouraged strategic research aimed at transforming multidisciplinary scientific
knowledge into real-life healthcare applications [1–3]. Successful applications of
compact systems are abundant, including DNA microarrays [4], chip-based poly-
merase chain reaction (PCR) [5], peptide and oligonucleotide libraries [6], drug
screening [7], cell culture [8], and even the concept of living systems on a chip to
replace tests using animals [9]. This development demonstrates a growing trend in
IVD tests involving versatile and miniaturized lab-on-a-chip (LOC) microsystems
that can integrate precise fluid handling, complicated sample processing and signal
detection, and readout systems for diseases monitoring, determination of the state of
health, or infection detection in order to cure, mitigate, treat, or prevent disease or its
sequelae [10]. Advances in bio- and nanotechnology continue to expand the science
and relevancy of translational medicine by expanding the scope and capacity of IVD
tests [11, 12]. Examples adopting LOC concepts for IVD applications can be easily
found in commercial products such as i-STAT (Abbott Point of Care, Princeton, NJ),
Dakari CD4 (Dakari Diagnostic, Cambridge, MA), Alere Triage MeterPro (Alere,
Waltham, MA), and Piccolo Xpress (Abaxis, Union City, CA), which adopt micro-
electronic and microfluidic components to create advanced IVD platforms [13].
In addition to conventional analytical materials or microfabrication substrates,
such as glass and silicon, polymeric materials have been identified as a good alter-
native owing to their mechanical flexibility, lightweight, mass fabrication capacity,
low cost, and multiple chemical/physical properties based on different grades [12,
14]. Therefore, they gradually become one of the major developing trends for IVD
systems. The most common polymeric materials used to fabricate chip-based IVD
devices include PDMS, COP, PMMA, PC, and PS [15–17]. A chart summarizing
material properties for polymeric IVD device fabrication is provided in Table 2.1.

© Springer International Publishing Switzerland 2016 15

C.-M. Cheng et al., In-Vitro Diagnostic Devices, DOI 10.1007/978-3-319-19737-1_2
 16

Table 2.1  A chart summarizing material properties for polymeric IVD device fabrication
Polymer Acronym Tg (°C) CTE Water Solvent Acid/base Biocompatibility Optical transmissivity
(10−60C−1) absorption (%) resistance resistance Visible UV
Polydimethylsiloxane PDMS −125–122 300–310 0.03 Poor Good Excellent Excellent Excellent
Cyclo olefin polymer COP 70–163 60–70 0.01 Good Good Excellent Excellent Good
2

Cyclic olefin copolymer COC 80–180 60–70 0.01 Good Good Excellent Excellent Good
Poly(methyl methacrylate) PMMA 100–122 70–150 0.3–0.6 Good Good Excellent Excellent Good
Polycarbonate PC 140–148 60–70 0.12–0.34 Good Good Excellent Excellent Poor
polystyrene PS 92–106 10–150 0.02–0.15 Poor Good Excellent Excellent Poor
CTE coefficient of thermal expansion
The variance of these parameters is based on the different grades of polymer
Polymeric-Based In Vitro Diagnostic Devices
2.1 Overview 17

In this chapter, the polymeric material-based IVD systems and the potential IVD sys-
tems adopting LOC concepts will be introduced. Rigid polymers such as PMMA,
PS, and PC can be used to fabricate less deformable structures. However, their capac-
ity is limited in some lab-on-a-chip applications due to their lower optic transmission
and weak organic solvent resistance. Here, we will mainly focus on the properties of
two different polymeric materials, PDMS and COP, which are commonly used for
new-type IVD prototyping and frequently potential product applications.

2.2 Selection of Polymer Materials

2.2.1 Polydimethylsiloxane

Polydimethylsiloxane (PDMS, Dow Corning Corporation) is probably the most

popular polymer due to easy fabrication and bonding for prototyping and test-
ing. PDMS is a commercially available silicone rubber. The physical and chemi-

low loss tangent (δ ≪ 0.001), flexibility (shear modulus ~0.25 MPa and Young’s
cal properties of PDMS include low glass transition temperature (≈−125 °C),

modulus ~0.5 MPa), high dielectric strength (∼14 V µm−1), reasonable tempera-
ture variations (thermal expansivity α ≈ 20 × 10−5 K−1), wide temperature opera-
tion range (from −100 °C up to +100 °C), and highly optically transparent from
the UV region to the NIR which make it an excellent candidate for optic sensing
[18–21]. It is intrinsically hydrophobic with a water contact angle of ~110°, but
the surface can be modified by oxygen plasma treatment to become hydrophilic. It
can adhere irreversibly, after oxidation, to many different types of substrates [22,
23]. Except at extreme pH values, it has low chemical reactivity. It is characterized
by a non-toxic, biocompatible nature with high permeability to O2 and CO2 that
facilitates cell culturing in lab-on-a-chip fashion [24]. Even some drawbacks such
as the stiffness are not as strong as other polymeric materials, surface swollen in
organic solvents, and most surface treatment results are often unstable over time
possess some limitation [25], other properties are well suited for IVD applications.

2.2.2 Cyclic Olefin Copolymer

As a substrate material for IVD tests, several rigid polymer materials including PC,
PMMA, PS, and COP have been considered. Among them, the unique properties
of COP, including high resistance to chemicals, high biological compatibility, high
transparency in the visible and near-ultraviolet regions of the spectrum, low auto-
fluorescence, low water absorption, low oxygen and moisture permeability, and
low gas emissions, make it a strong candidate material for IVD formats [26–28].
COP is highly resistant to acids, alkaline agents, and polar solvents. It is only
attacked by nonpolar organic solvents, such as hexane and toluene [29]. This
18 2 Polymeric-Based In Vitro Diagnostic Devices

chemical resistance is critical for chip-based sample extraction and separation sys-
tems that require multiple washing, loading, elution, and recondition processes
[30, 31]. It is also crucial for various bioprocessor, biosensing, monitoring, and
screening applications that require multiple chemical reaction processes or a harsh
operating environment [32].
The outstanding optical properties of COP allow it to be fabricated as a wave-
guide and lens material [33]. COP has a high optical transparency over a wide
wavelength range from 300 nm to 1200 nm, a large Abbe number, a high refractive
index, and a low birefringence, so it can be easily integrated with extra optic sys-
tems for sensing. Moreover, in the near-UV region, the transmittance is higher for
COP than PMMA, PC, or PS, so it can be used for surface modification by photo-
chemistry synthesis [33, 34]. Another vitally important property of COP is its low
autofluorescence, which lowers the background noise when fluorescence detection
is employed. The autofluorescence of COP is higher than that of glass or PDMS,
but in the same order of magnitude as that of PMMA or PC [33, 35].
The water absorption capacity of COP is about four times less than that of PC
and about 10 times less than that of PMMA [36, 37]. This low water absorption
provides excellent dimensional stability under a variety of environmental condi-
tions and limits potential solution concentration changes resulting from reagent
evaporation and/or absorbance when long processing times are required.
COP is based on ethane and cyclic olefin monomers. Various COP materials
are commercially available under brand names including TopasPAS (TOPAS,
Florence, KY), APEL (Mitsui Chemicals, Tokyo, Japan), ARTON (Japan
Synthetic Rubber, Tokyo, Japan), Zeonex, and Zeonor (ZEON Corporation,
Tokyo, Japan). The difference between them is depending on the cyclic mono-
mer and the polymerization process used during synthesis [38, 39]. COP products
from Topas and Apel are based on the chain copolymerization of cyclic mono-
mers with ethene, and Arton, Zeonex, and Zeonor are ring-opening metathesis
polymerization of cyclic monomers followed by hydrogenation [26, 40]. Even the
same brand, different grades with different glass transition temperatures (Tg). Tg
increases with a higher cyclic olefin content, so some COP grades possess a higher
glass transition temperature than PMMA, PC, and PS [41]. This makes it possible
to use certain grades of COP materials in applications exposed to higher tempera-
tures without the risk of deformation and various thermal bonding or micromold-
ing processes [42, 43].

2.3 Fabrication of Polymer Devices

2.3.1 Structure Formation

Versatile fabrication methods are available to structure polymeric materials to

form particular geometry for the manufacture of IVD test devices. Laser ablation
and micromilling are direct structuring methods suitable for fast prototyping with
2.3 Fabrication of Polymer Devices 19

minimal preparation. Injection molding, also available, is an ideal fabrication pro-

cess for mass production of commercial available products. Soft lithography, hot
embossing, and nanoimprint lithography are more appropriate replication methods
for low-cost and laboratory-based production.

2.3.1.1 Soft Lithography

Photolithography continues to be the dominant technology in semiconductor fab-

rication [44]. As the most important and profitable microfabrication technique,
it has contributed to the development of IVD applications including the fabrica-
tion of DNA arrays in the late 1980s [45]. However, this technique has a num-
ber of limitations, such as an intrinsically expensive fabrication environment and
costly equipment, surface modification difficulties, and obstacles to plain surface
morphology manipulation that decrease its application in biomedically relevant
research fields.
Soft lithography is a technique based on microstructure formation, molding,
and embossing to obtain a reverse elastomeric stamp (Fig. 2.1) [46, 47]. These
techniques were developed as an alternative to photolithography. No specific labo-
ratory environment is required, and the process does not involve expensive equip-
ment. Soft lithography is a non-photolithographic strategy based on self-assembly
and replica molding for carrying out micro- and nanofabrication. It can con-
tinuously create large three-dimensional features that can be used in an ordinary
laboratory without the need for clean room facilities. In its initial steps, soft lithog-
raphy relies on the use of photolithography to generate a master used for replica-
tion. Once the master is fabricated, the fabrication tasks can be performed outside
of a clean room via printing or molding procedures. A large number of pattern-
ing techniques such as replica molding [48], microtransfer molding [49], sol-
vent-assisted molding [50], micromolding in capillaries [47], phase-shifting edge

Fig. 2.1  The fabrication (a)

of PDMS slab using soft
lithography. a, b Master is
first formed by spincoated
photoresist on a silicon wafer
followed by photolithography (b)
processes. c PDMS mixture
is then poured on the
master and cured thermally.
d The peeling-off layer (c)
of PDMS slab has invert
microstructures to the master

(d)
20 2 Polymeric-Based In Vitro Diagnostic Devices

Fig. 2.2   Schematic illustration depicting the procedure for a replica molding (RM), b micro-
contact printing (μCP), and c solvent-assisted micromolding (SAMIN) [58]

lithography [51], decal film transfer lithography [52], nanotransfer printing [53],
microcontact printing [54], nanoskiving [55], and dip-pen nanolithography [56,
57] have been developed. Soft lithography is a cost-effective option that allows for
the use of adjustable surface chemistries, requires a minimal laboratory environ-
ment, and is highly compatible with biological applications including cell biology,
microfluidics, and a variety of lab-on-a-chip systems.
In the following section, three commonly used soft lithographic techniques are
introduced, including replica molding, microcontact printing, and solvent-assisted
micromolding (Fig. 2.2) [58].
Replica molding is a process that transfers a pattern from a rigid or elastomeric
master mold into another material via solidification of liquid poured into the mold.
This new method for fabrication of PDMS-based IVD devices for prototyping [22]
has proven to be particularly suitable for numerous biomedical device applica-
tions [59–62]. Because the production of PDMS microstructures is simple, it can
be readily used to make prototype devices and full-function integrated systems
[10, 63]. It is also an attractive process for nanofabrication of devices with lat-
eral dimensions smaller than 100 nm [64]. This technique has also been used to
micropattern biocompatible polymers including epoxies, polyurethanes, polyethyl-
ene glycol (PEG), agar, and agarose for isolating and culturing bacterial cells.
Microcontact printing (µCP) is a large-area (>cm2) patterning technique used
on functional organic surfaces. The process is similar to using a common stamp to
transfer ink from an ink pad to a piece of paper. In this process, a mold is stained
with a chosen material, e.g., small biomolecules, proteins, polyelectrolytes, or
suspensions of cells, and this material is transferred to the substrate surface when
contact is made between the substrate and the protruding features of the stamping
mold. µCP has been successfully used to print precise patterns of axon guidance
molecules as a cell growing template for growing chick retinal ganglion cell axons
[65]. It also allows for the engineering of surface properties via molecular-level
detailed adoption of the self-assembled monolayer (SAM) technique on the sub-
strate when PDMS stamps stained with alkanethiols (SH-(CH2)n-X) are used to
microcontact print on surfaces of gold, silver, palladium, platinum, or other metals
2.3 Fabrication of Polymer Devices 21

[66]. Formation of alkanethiolate SAMs include thin-film, physical vapor deposi-

tion on silicon, mica, glass, or plastic materials [67]. Patterned SAMs are valuable
for studying the role of spatial signaling in biosensing and cell biology by control-
ling the molecular structure of a surface in contact with cells and proteins on dif-
ferent platforms [56].
Solvent-assisted micromolding (SAMIN) is similar to replica molding and
is based on molding or embossing with an elastomeric stamp. In this proce-
dure, an elastomer mold is wetted with a solvent before the conformal contact is
made between the elastomer mold and the substrate. The liquid solvent fills the
recessed regions on the elastomer mold contact surface, which minimizes the
area of the liquid/vapor interface and maximizes the solid/liquid interface. As a
result, nanoscale structures can be produced in various soft materials over large
areas (>cm2 with 100 nm features). This process can also be combined with selec-
tive etching and liftoff processes to transfer into metals that can then be used as
substrates for various biomedical sensing platforms such as electrochemistry, [57]
surface plasmon resonance [68], optical diffraction [69], and surface-enhanced
Raman scattering [70].

2.3.1.2 Injection Molding

Injection molding is a scalable strategy for manufacturing thermoplastic materials

with features on the order of micrometers and above. It is one of the most common
techniques for the fabrication of polymeric products, since it is highly adaptable
for mass production [71, 72]. The process involves initially feeding polymeric pel-
lets into an injection molding machine hopper and then applying high temperature
to melt the pellets before the mass is injected into a mold and high pressure is
applied. This constant packing pressure is applied for a brief time before the poly-
meric material and mold are cooled and the manufactured piece is demolded.
The quality and fidelity of the replicated structures depends to a great extent
on the master and the fabrication processes. The high pressure and temperature
ranges used when molding limits the use of silicon, glass, resists, and other pol-
ymers as mold material, so metal materials are commonly chosen [73]. Those
molds with micrometer resolution for LOC applications are usually fabricated
using standard photolithography techniques followed by electroplating to prolong
the mold’s lifetime [74–76]. A master mold can withstand more than 200 cycles
without severe deformation [77].
Many parameters, including injection speed, mold temperature, and structure
geometry in the injection molding process, have a direct effect on polymeric rep-
lica quality and usability. Because injection masters may incur structural disrup-
tion after a number of cycles, surface replication patterns are influenced not only
by pressure distribution inside the mold but also by internal structural deformation
[78]. This can often be resolved, and master cavities can be filled more effectively
by using higher injection temperatures that produce better flow behavior [79].
22 2 Polymeric-Based In Vitro Diagnostic Devices

Fig. 2.3  a Image of injection molded polymer chip and b scanning electron micrograph (SEM
image of the nanochannels on the injection molded portion of the device [80]

Injection molding has been used to produce LOC systems for on-chip liquid
chromatography and DNA bar coding systems [43, 80]. However, this process can
be challenging because of the large aspect ratios required, low surface roughness,
high flow pattern resolution, and stresses experienced by the replica. Replication
of nanostructures into polymer surfaces has been successfully achieved and had
received a good deal of attention (Fig. 2.3) [80, 81]. The main limitation of this
process lies in the grain size of the master, which limits its ability to successfully
replicate smaller structures.

2.3.1.3 Hot Embossing

Hot embossing is a technique that employs a polymeric sheet pressed onto a

microstructured mold or wafer heated above the glass transition temperature
(Tg), followed by demolding at approximately Tg −50 °C [42, 82, 83]. Figure 2.4
depicts a schematic of the hot-embossing process [84]. The applied pressure
to hold the template and sheet, pressed time, and operation temperature are the
most important factors influencing the end quality of the embossed structures on
the chip surface. An anti-sticking layer is usually used to provide good fidelity for
large-area embossed structures. The chip or wafer format polymeric substrate can
be acquired commercially or formed by injection molding or by heating polymer
pellets above Tg [85].
Hot embossing is generally done using lower fabrication temperature ranges
and lower pressure compared to injection molding. Based on the lower pressure
and temperature used, silicon, copper, nickel, stainless steel, and even polymer
masters have been used as template material [85, 86]. Among them, SU-8 pho-
toresist has also been used as a master material through standard photolithography
for embossing polymeric materials under conditions similar to the ones used with
metal templates, and an aluminum coating can be applied to facilitate substrate
demolding from the template [87]. To fabricate high aspect ratio structures, an
2.3 Fabrication of Polymer Devices 23

Fig. 2.4  The schematic hot embossing process. The total force (F) required to emboss a thermo-
plastic polymer depends on the polymer’s viscosity, contact area of the stamp features with the
polymer (c), surface area of the entire stamp (C) and temperature [84]

anodic aluminum oxidation nanostructured membrane and cylindrical pillars can

be used [88, 89].
The distance that the substrate must flow toward the template in the hot-
embossing process is smaller than that for high-temperature injection molding,
which leads to reduced stress and shrinkage effects during operation [90]. Lower
mold temperatures and cooling rates lead to production of more fragile structures
with higher aspect ratios than those achieved with injection molding.

2.3.1.4 Nanoimprint Lithography

Nanoimprint lithography (NIL) is a non-conventional lithographic technique for

low-cost, high-resolution patterning of polymer nanostructures. During the NIL
process, which is performed in a vacuum chamber with two parallel templates
pressed together, the substrate is first heated to a temperature above the Tg and
then, the templates are pressed against the substrate at a pressure for a period of
time. After imprinting, the substrate is cooled down, under constant pressure, to
a temperature lower than Tg to avoid deformations and demolding is performed
at room temperature. In contrast to traditional lithography, which uses pho-
tons or electrons to modify the chemical and physical properties of the resist to
achieve high definition patterns, NIL relies on direct mechanical deformation of
the molded material to achieve high-resolution patterning (Fig. 2.5) [91]. In hot
embossing, only a small portion of the substrate surface is embossed into the tem-
plate mold; however, because NIL uses most or all of the polymeric film thickness
for pattern production, the residual polymer layer left after substrate imprinting
is quite thin and may be nonexistent [92]. O2 plasma in a standard reactive ion
etcher system is usually used to etch the remains of the residual layer. The etching
Other documents randomly have
different content
 10 THE PSYCHOLOGY OF ADVERTISING read descriptions
and see pictures of it and then I would think of it (have ideas of it)
in terms of touch, weight, smell, and taste which were taken from
former experiences in which similar objects were present to my
senses. Whether we think by means of perceptions or by means of
ideas, the original material of thought and the forms of thought
come to us in sensations. The original, easiest, and surest method of
acquiring knowledge is through perceptions, in which the sensations
play a leading part. In many instances the object of thought cannot
be present to the senses and, furthermore, the processes of thought
are made more rapid by substituting symbols for the original. Thus,
early in the history of the race, a spoken language was developed in
which spoken words were symbols for objects of thought. Later, a
pictorial writing was invented in which crude portraits were made to
symbolize objects. The latest products of civilized humanity in this
direction are, first, more perfect portraits land, second, a form of
printed language in which the original symbolic spoken word is
represented by a sym]t>ol. This second form is the most convenient
and is the one in ordinary use, but it should be observed that our
printed words are nothing but symbols of symbols. The printed word
is an uninteresting thing in itself and is only used because it assists
perception on account of its simplicity and ease of manipulation. It is
easy to describe a scene or a commodity and to reduce the
description to printed form that will be accessible to thousands. It
would be extremely difficult to deliver the scene and the commodity
directly to these same people. The description and illustration are,
however, not so clear, distinct, and interesting as is the original thing
described The great danger with the printed
 PERCEPTION 11 symbol is that it will lose in perspicuity and
interest what it gains in convenience. The printed word has almost
no interest for us in itself. It becomes interesting only in so far as it
symbolizes interesting things to us. The more the printed page has
to say and the easier it is for us to interpret it, the more interesting
it becomes. Whether fortunately or unfortunately, the advertiser is
compelled to rely on symbols in exploiting what he has to offer. He
cannot, ordinarily, provide the possible customer with that which he
has to offer and thus allow him to become acquainted with the
goods in the normal and direct way. He is compelled to substitute
the symbol for the thing symbolized. He has a choice between two
kinds of symbols — printed words and pictorial illustrations. The first
form of writing was picture writing, but was abandoned because it
was not so convenient as are the phonetic characters now in use.
Picture writing could not be written or read so easily and quickly as
the writing in the characters now in use and it was therefore
discarded. According to the standard of ease of interpretation, all
forms of type must be judged. Type forms must not be regarded as
a production of artistic demands, but as a product of the demands of
convenience. Hundreds of styles of "artistic type" have been brought
forth, but they have not remained in use, for they are confusing to
the eye and are not artistic in the full sense of the term. Those
forms of type and of illustration best perform their proper functions
which are so easy of interpretation that they are not noticed at all.
There is no advantage in emphasizing the symbol, but there is a
great advantage in emphasizing the thing symbolized. In using
printed forms, the adver 
 12 THE PSYCHOLOGY OF ADVERTISING tiser supplies a
very small part to the total idea whicli lie desires to create, and he
should therefore make this little mean as much as possible. A series
of experiments were carried on to determine whether white or black
type made the more attractive display in magazine advertisements.
Experiments were made with over five hundred persons. The
background for the white type was gray in some cases, but in most
cases it was black. The results show that the ordinary reader is more
likely to notice display type which is black than a display type of the
same sort which is white. A series of laboratory experiments were
made on the same subject. Specially prepared pages were shown for
one-seventh of a second. On part of the sheets black letters on
white background and white letters on black background were
shown. In other cases one half of the sheet had a black background,
with words in wiiite type, and the other half of the sheet had a white
background with words in black type. Scores of cards were
constructed in which all the possible combinations of white and black
were made and shown to a number of persons for such a short
space of time that no one could perceive all there was on any sheet.
Under these circumstances the subjects s;aw what first attracted
their attention and what was the easiest to perceive. The final
results showed that the black letters on a white background were
seen oftener than the white type on a black background. It seems
quite certain that, other things being equal, tliose advertisements
will be the most often read which are printed in type which is the
most easily read. The difference in the appearance of the type in
many cases may be so small that even persons experienced in the
 PERCEPTION 13 choosing of type may not be able to tell
which one is the more legible, and yet the difference in their values
may be great enough to make it a matter of importance to the
advertiser as to which type he shall use. If the matter of the proper
use of type is of importance to the advertiser, it is even more
important that he should make a wise use of the illustration, which is
the second form of symbol at his disposal. The illustration is
frequently used merely as a means of attracting attention, and its
function as a symbolic illustration is disregarded. In a few cases this
may be wise and even necessary, but when we consider the value of
an illustration as a symbol, we are surprised that illustrations are not
used more extensively as well as more judiciously. The first form of
writing, as stated above, was picture writing, and the most simple
and direct form of graphic representation is through the picture and
not through the printed word. At a single glance we can usually read
about four words; that is to say, the width of perception for printed
words is about four. At a single glance at an illustration we can see
as much as could be told in a whole page of printed matter. The
width of perception for Illustrations is very much more extensive
than it is for printed forms of expression. The illustration may
perform either one or both of two functions. It may be a mere
picture used to attract attention or it may be an "illustration" and a
real aid to perception by assisting the text to tell the story which is
to be presented. In the first case it would be called an irrelevant
illustration; in the second case it is relevant. There have been
several investigations carried on to determine the relative attention
 14 THE PSYCHOLOGY OF ADVERTISING value of relevant
and irrelevant illustrations. Although the results thus far reached are
not so decisive as might be desired, yet it seems certain that the
attention value of relevant illustrations is greater than had been
supposed and that the irrelevant ^'picture" is frequently not so
potent in attracting attention as a relevant illustration would be.
Under these circumstances it seems that, in general, the illustration
in an advertisement should have the double function of attracting
attention and assisting perception. Which one of these functions is
the more important might be a profitable question for discussion,
but when these two functions can be united in the same illustration,
its value is enhanced twofold. Irrelevant illustrations are produced
merely because they are supposed to attract attention, when in
reality they may attract the attention of no one except the person
who designed them and of the unfortunate man who has to pay for
them. Similarly there are many illustrations produced and inserted in
advertisements because they are supposed to assist the perception.
They are supposed to tell the story of the goods advertised and to
be a form of argumentation. The designer of the illustration and one
familiar with the goods knows what the picture stands for, and so for
him it is a symbol of the goods and tells the story of the special
advantages of the goods. To one unacquainted with the illustration
and with the goods advertised, the illustration is no illustration at all.
When we want to teach a child the letters of the alphabet, we do not
secure some "sketchy" and artistic looking letters,, but we secure
those which are simple in outline and of a large size. We choose
those which make a very decided sensation, for in that way we help
determine the perception. When the child becomes
 PERCEPTION 15 more familiar with the alphabet, he can
read small letters and those which are not printed so plainly. In
forming perceptions there must at first be a large element furnished
by sensation, whether the perception be formed from an object
directly or indirectly from a symbol. Those who forget this principle
are likely to construct illustrations which do not illustrate. Their
symbols are only symbols for those who are well acquainted with the
goods advertised. As an example of this sort of illustrations we
reproduce herewith an illustration from magazine advertising. EP.C.
WAX is the l>est and most economical Laundry Wax sold your The
kind that keeps' the iron CLEAN (aSMOOTH Put up in little wooden
tubes with an automatic handle that keeps the wax in position and
prevents waste The neatest and nicest •way that wax can be used
for ironing purpose ^ AND for S two-cent stamps we will send you
2* sticks to try -After that they can be had from dealer 'cause Z
never satisfy FLAME PROOF CO. NEW YORK No. 1
 16 THE PSYCHOLOGY OF ADVERTISING This advertisement
for F. P. C. wax (No. 1) seems to be an attempt to tell a great deal
about the goods by means of an illustration. It took me some time
to translate it, and after I had interpreted it as far as possible, I
showed it to some ladies who were magazine readers. None of them
had ever taken the pains to figure it out. One of them thought that it
was an advertisement of Bibles. When my attention was called to it,
I saw the resemblance between the cut as a whole and the cover of
an ordinary Bible. The white space is evidently intended to look like
the bottom of an iron and the border containing the words "F. P. C
Wax" is intended for a cut of a stick of the wax. None of the ladies
had interpreted the cut in that way, but when their attention was
called to it, they agreed with me that that was probably what the
"artist" had intended. We were unable to interpret the white dots
and the heavy black border. To those familiar with the advertisement
the sensation aroused by the cut is sufficient to produce the desired
perception. For all others the sensation is not sufficient to call up the
necessary elements to complete the perception and it has no more
meaning than a Chinese puzzle. It has nothing which it seems to be
trying to tell to those who turn over the pages of the magazine, and
so does not attract their attention. We notice those illustrations
which have something to say and say it plainly. We disregard in
general those things which do not awaken in us a perception. The
sensation which does not embody itself into a perception is of such
little interest to us that we pay no attention to it at all. The
advertiser desires to produce certain perceptions and ideas in the
minds of the possible customers. The material means with which he
may accomplish
 PERCEPTION 17 this end are printed words and
illustrations, which in the first instance awaken sensations; these in
turn embody themselves into perceptions and ideas. These
sensations seem so unimportant that they are frequently No. 2
forgotten and the place which they are to take in forming the
desired perceptions and ideas is disregarded. This second
advertisement of F. P. 0/ wax (No. 2) appeared several months later
than the one given above, and is inserted here to illustrate how an
advertisement may be improved in the particular point under
discussion. The newer cut is really an illustration. It
 18 THE PSYCHOLOGY OF ADVERTISING helps perception
by giving a sensation which is more decided and more easily
interpreted. It furthermore attracts attention and tells the story
better than could be done by any text. The advertiser is so familiar
with what he has to offer that he cannot appreciate the difficulty the
public has in getting a clear and complete perception by means of
his advertisements of the goods advertised. It is almost impossible
to err on the side of clearness. A sketchy illustration may appear
artistic to the designer, but there is danger that it will be regarded as
meaningless scrawls by the laity, and so it will not receive a second
thought from them. The text and the illustration should, first of all,
be clear and should in every way possible assist the mind of the
possible customer in forming a correct idea of the goods being
exploited.
 APPERCEPTION 19 III APPERCEPTION Anatomy is the
science which divides the human body into its constituent parts, and
is a completed science when it has all of these parts correctly
described and labeled. Physiology is the science which describes and
explains the different functions of the human body. It supplements
anatomy by showing the function of each of the bones, muscles, and
organs, and by showing their mutual relations. In anatomy we divide
the body into distinct divisions, and in physiology we discover
different functions. We often try to think of mind after the analogy of
the body, and by so doing are led into confusion. The attempt has
been made to divide the mind into a definite number of separate
faculties (anatomy). The function of each faculty has been described
as something quite different from the other faculties, and an attempt
was made to define these faculties exactly and to describe their
functions completely (physiology). The attempt has failed and has
been abandoned. The mind is not a bundle of faculties. It is not
composed of memory, reason, association, etc., but it is a unit which
remembers, reasons, feels, etc. No one function is carried on to the
exclusion of all others at any one time. During all of its conscious
existence the mind feels, knows, wills, etc., but at certain times it is
employed in reasoning more than at others, and at one time it may
be feeling more intensely than at others, but no one function ever
totally occupies the field.
 20 THE PSYCHOLOGY OF ADVERTISING When the mind
recognizes an event as having occurred in tlie past, it is said to
remember, but feeling, attention, and association of ideas may have
entered into this process of memory. No one mental process is a
thing existing apart and independent of other processes. The
anatomical method can never be applied to the mind. The functions
of the mind are not independent activities of the mind, but in every
function memory, perception, suggestion, and many other functions
play a more or less important part. We have no "apperceiving"
faculty which is to be distinguished from all other faculties, and
which carries on an independent process. The mind does act in a
particular and well-known manner, which we have called
"apperception." The term has been used for two centuries, and is
applied to a well-known process, or function, of the mind which is of
great practical and theoretical importance. It includes sensations,
perceptions, assimilation, association, recognition, feeling, will,
attention, and other actions of the mind, and yet is a very simple
and well-known process. It can best be understood if discussed and
illustrated from its various aspects. The first thing to be said about
apperception is that it is the act of the mind by which perceptions
and ideas become clear and distinct. I may look at my ink bottle on
the middle of the table. I see it very clearly and distinctly. I can also
see, at the same time, other objects on the table, and even some
which are not on it at all. As long as I continue to look at the ink
bottle the objects distant from the table are not visible. The ink
bottle is very clear and the objects near it are comparatively so ;
those a few feet away are very indistinct or entirelv invisible. I am
said to apperceive the bottle,
 APPERCEPTION 21 but to perceive the more distant
objects. Certain parts of the bottle are not noticed particularly, while
some of the objects on the table stand out plainly. It is quite evident
that ^^clearness" does not draw a set line between the various
objects, but there are all grades of clearness, from the most clear to
the most obscure. We feel that the mental process connected with
the ink bottle and that connected with the other objects are different
and yet there is an uninterrupted gradation from one to the other.
When considered from this point of view apperception is simply an
act of attention, for what we attend to becomes clear and distinct to
us, while that which is not attended to remains indistinct.
Furthermore, there are all degrees of attention. Certain things
demand our greatest attention, while others are entirely
disregarded. Most things, however, are of the intermediary class. We
pay a certain amount of attention to them, but they might easily
receive more or less. Some things catch our attention so slightly (are
so slightly apperceived) that we are not aware that we have noticed
them at all. I did not know that I had ever noticed the walls of the
barber shop which I patronize, but as soon as I entered it recently I
knew that changes liad been made, and I missed certain details
which I had frequently seen, but to which I had paid so little heed
that they were merely perceived and could not be said to have been
apperceived at all. The second thing to remark about apperception is
that it is more than mere attention. It is attention of a particular
kind. Our attention to an object or event is an act of apperception if
the attention is brought about by means of the relationship of this
object or event to our previous experience. Apperception has been
defined as the bringing to hear what has been retained of past
 ^22 THE PSYCHOLOGY OF ADVERTISING experience in
such a way as to interpret^ to give weight to the new experience.
This aspect of apperception has been most clearly brought out in the
following quotation from Dexter and Garlack : "A child who has not
learned any physiology, and who has not previously looked through
a microscope, looks at a drop of blood under the microscope. He
probably says that he sees nothing. "Another child who has, we will
suppose, studied botanical sections under the microscope, looks at
the same drop of blood and says that he sees some small round
bodies. "A third child who has learned a little physiology, looks
through the microscope, recognizes the small round bodies as
corpuscles, notes that the majority are reddish, looks for and
perhaps finds a white corpuscle, and so comes to the conclusion that
it is a drop of blood that he sees. "In the three instances everything
is the same except the children. The differences in the results of the
acts of observation must be due to the differences in the minds of
the children. The reason that the third child saw more than the other
two was that he was fitted by previous training to see more. In order
that we may see a thing properly it is not sufficient that rays of light
should come from the object to the eye and nerve vibrations travel
along the optic nerve to the brain. The mind must be in a position to
interpret, to understand these vibrations. To sensations coming from
without the mind adds imagination (i.e.^ image-making) working
from within. This combination of action of object on mind and the
reaction of mind on object is known as apperception.^^ The third
thing to notice about the process of apper 
 APPEECEPTION 23 ception is that it increases our
knowledge by gradually adding new elements to pur previous store
of experience. In the use of the microscope, as cited above, ^acli
child added to its store of knowledge in proportion to the amount of
previous training which could be brought to bear at this point. The
first child had had no previous training in this or in any related work,
and so was unable to profit by this experience. He did not focus his
eye correctly, and could not direct his attention to what the third
child saw. An object, event, or situation which has no relation to our
previous experience fails to attract our attention, — is not
apperceived, — makes no impression on us, and adds nothing to our
store of knowledge. Nothing is regarded worthy of our consideration
which does not relate itself to our previous experience. In fact, we
can imagine nothing which is out of relation to all our previous
experiences. Things and events are only significant in so far as they
signify relationships which we know. The slight difference between
the letters "O" and "Q" is immediately noticed by us, but would not
be seen by any one unfamiliar with our alphabet. There are many
important characteristics about the Chinese alphabet which we never
observe, because they mean nothing to us. They are unimportant for
us because they do not unite themselves with our previous stock of
ideas. We interpret all things by our own standards (our stock of
ideas) — we observe only those things which have significance for
us, we increase our store of ideas not by adding new and
independent ones, but by uniting the old with the new. We are not
capable of forming entirely new ideas, but must content ourselves
with adding new elements to our stock in trade. All our so-called
new ideas are composed very largely of old elements.
 24 THE PSYCHOLOGY OF ADVERTISING The practical
importance of this subject for the advertiser is found in the three
aspects of the process as discussed above. In the first place, some
advertisementsnever stand out clearly and distinctly in the minds of
the possible customers. We may turn over the pages of a magazine
and see every advertisement there, but our seeing may be of the
sort of those of whom it was said, "having eyes they see not.'' I
frequently turn over £he pages of publications and direct my eyes
toward advertisements and hold them there long enough to have
noticed all the striking characteristics of them, and yet in ten
minutes afterward I do not know that these particular
advertisements are in the publication at all. I had perceived them,
but had not apperceived them. The designers of these
advertisements had not been successful in concentrating my mind
on any particular thing which had a special reference to my previous
experience, and which would therefore be apperceived by me. We
cannot apperceive a large number of things at the same time. An
advertisement which is constructed upon the principle that all parts
of it should be attractive at the same time will so divide the attention
that no part of it will stand out prominently, and so it will not be
noticed at all. A superfluity of details should be strenuously guarded
against in both the text and the illustration. If a single point of an
advertisement is apperceived it serves as an opening wedge for the
entire advertisement. If, however, there are too many details the
attention may be so distracted that none of it will be apperceived,
although it may all be seen (perceived). The things which we
perceive do make a slight impression on us, but they are so
unimportant in comparison with the things that we apperceive that
we may almost disregard them entirely.
 APPERCEPTION 25 The second point for the advertiser to
consider is that the apperception value (identical with attention value
in this case) of the advertisement does not depend so much on what
the reader receives from the advertisement, but what he adds to it.
Your advertisement and all other printed matter is composed of a
few straight lines and" a few curved ones, of a few dots, and
perhaps one or more colored surfaces. These, when seen, cause a
sensation of sight, but that is the smallest part of the result of your
advertisement. These visual sensations are immediately enforced by
the previous experience of the reader. The value of your
advertisement depends almost entirely on the number and kind of
former experiences which it awakens. The advertisement is not a
thing which contains within itself the reason for its existence. In and
of itself it is perfectly worthless. The aim of the advertisement is to
call forth activity in the minds of its readers — and, it might be
added, action of a particular sort. The advertisement which is
beautiful and pleasing to its designer, and which begets activity in
his mind, may be perfectly worthless as an advertisement. The drop
of blood in the microscope brought forth no activity on the part of
the first child who looked at it, as cited above. The child had nothing
in its former experience which was suggested by the appearance of
the drop of blood, and so it was not interpreted and was not
connected with the child's former life, and so made no impression on
him. That which happened to the children in looking through the
microscope happens every day to the readers of advertisements.
The same advertisement will call forth different amounts of activity
from different readers. Some advertisements have a meaning to
those who are well acquainted with them, and to such they tell their
story accurately and quickly.
 26 THE PSYCHOLOGY OF ADVERTISIIS^G To some readers
they tell a confused or erroneous story ; to others they have nothing
to tell at all. As an example of such advertisements we have
reproduced the advertisement (No. 1) of Whitman's chocolates. No.
1 This looks like a very neat advertisement, but it fails at the two
crucial points — ^it neither attracts attention nor assists in forming a
correct perception of the goods advertised. As a proof of this
statement it is but necessary to refer to the result obtained with this
advertisement in a series of tests recently made. The magazine
 APPERCEPTION 27 containing this advertisement was
shown to 516 yonng people between the ages of ten and twenty-
five. After they had looked at all the advertisements they were asked
to write down all the advertisements which they had noticed and
could remember. One girl remembered that she had seen an
advertisement of candy, but could not remember whose it was or
what the advertisement was. One boy remembered that
"Whitman^s candy'^ was advertised, but thought the advertisement
had the picture of a lady eating a piece of candy. The first of the two
probably referred to Huyler's advertisement (Huyler advertised in the
same issue) and the second certainly confused the two
advertisements. Besides these two none of the 516 persons noticed
the advertisement sufficiently to remember that it was there at all.
This second advertisement (No. 2) of Whitman's appeared in a later
issue of the same magazine. I have made no tests of this
advertisement, but feel sure that if the 516 had seen this instead of
the other advertisement a very large per cent, of them would have
noticed it and have remembered it. It attracts attention and tells
more at a glance than could be told in many well-formed sentences.
It would create a desire on the part of many of these young people
to send for or to purchase a box of such desirable looking candy. It
is an illustration which illustrates by helping perception, and it also
attracts attention because it has something to tell. The third thing
for the advertiser to observe in connection with apperception is that
advancement in knowledge is made by joining the new on to the old.
The pedagogical maxim of advancing from the known to the
unknown finds its justification here. It is very difficult to get the
public to think along a new line, because they cannot connect th^
new fact with
 28 THE PSYCHOLOGY OF ADVERTISING their previous
experience, i.e., they cannot apperceive it. This makes it very
difficult to introduce a new article on the market. Old firms find it
difficult to introduce a new brand, and new firms find it difficult No.
2 to get themselves noticed at all. Frequently firms have resorted to
questionable means to get the public even to notice them. It seems
to be impossible for them to get a hearing for the details of their
propositions until they have let the public become familiar with their
 APPERCEPTION 29 names and know who they are. The
promoters of Omega Oil have been severely criticised for their
goose, but the goose has introduced them to the public, and now
they are in a position to get a hearing and to present the arguments
for their commodity. It is quite possible that the expense of keeping
the goose before the public was an unnecessary luxury, but they
have been wise in not advancing their argument faster than the
public was willing to hear it. They have taken but one step at a time.
They first let the public know that there was such a thing as Omega
Oil, and they took great pains to make this new fact known, and in
doing this they were acting in accordance wdth the principles of
apperception. They first gave the public some experience of Omega
Oil, and then tried to get the public to interpret their arguments in
the light of tha*t previous experience. It is not always necessary or
even wise to attempt to present all the arguments for a commodity
at a single time. It is frequently wise to carry on an educational
campaign and to present single arguments. In this way the mind of
the possible customer is not crowded with a lot of new and
disconnected facts, but each argument has time to be assimilated
and to form a part of his experience, and is called up to strengthen
and impress each succeeding argument. In writing an advertisement
the public to be reached must be carefully studied. In exploiting a
new commodity the writer should ask himself what there is about his
goods which will fall into "prepared soil'^ on the part of the reader.
The reader must first be appealed to by something which he already
knows, and thus activity on his part is awakened, and this activity
may be made use of for presenting the new elements, which, if
presented at first, would have met with no response
Welcome to our website – the ideal destination for book lovers and
knowledge seekers. With a mission to inspire endlessly, we offer a
vast collection of books, ranging from classic literary works to
specialized publications, self-development books, and children's
literature. Each book is a new journey of discovery, expanding
knowledge and enriching the soul of the reade

Our website is not just a platform for buying books, but a bridge
connecting readers to the timeless values of culture and wisdom. With
an elegant, user-friendly interface and an intelligent search system,
we are committed to providing a quick and convenient shopping
experience. Additionally, our special promotions and home delivery
services ensure that you save time and fully enjoy the joy of reading.

Let us accompany you on the journey of exploring knowledge and

personal growth!

textbookfull.com