CHAPTER FOUR

CYTOLOGY AND HISTOPATHOLOGY

SPECIMENS(By Mitiku Solomon)

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 COURSE OBJECTIVES
At the end of this chapter you will be able to:
• Understand different types of histopathology
specimens.
• Know collection and handling of histopathology
specimen
• Understand d/t types of histopathological
specimens and how they are collected.
• What should be considered during
histopathological specimen collection.
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 Specimen collection and handling
• In histopathology laboratory, proper biopsy
preparation and correct diagnosis depend on
proper tissue collection and handling.
• Proper tissue handling avoids the grave
mistakes committed by incorrect labeling and
mixed up of the tissue biopsy and request form.
• Another problem is the tissue placed in narrow
mouthed bottle that necessitates breaking of
the container in order to take out the tissue.
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 Specimen collection and…
• Sometimes the tissue biopsy may be placed in
inadequate amount of formalin.
• Laboratory personnel must have appropriate
knowledge of proper tissue handling.
• Upon reception of tissue specimens, the
following points should be noted:
• Request form submitted should be matched
with the tissue specimen.

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 Specimen collection and…
1.Tissue and organ specimens should be
submitted in a wide- mouthed capacious
container in order to avoid distortion.
3. The tissue or organ submitted should be
immersed with adequate amount of fixative
4. Label the tissue or organ specimen properly
5. Record all the relevant information on the
laboratory record book.

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 Specimen…
• The histological specimen can be prepared as :
• 1. Whole mount
• 2. Sections
• 3. Smears.
 1. Whole mount: These are preparation entire animal eg. fungus,
parasite. These preparations should be no more than 0.2-0.5 mm in
• thickness.
 2.Sections- The majority of the preparations in histology are
sections. The tissue is cut in about 3-5 mm thick pieces processed
and 5 microns thick sections are cut on a microtome. These are
then stained and permanently mounted.
•

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 Specimen…
• Microtomes are special instruments which
have automatic mechanism for cutting very
thin sections. To cut the sections on the
microtome;
• the tissue must be made hard enough to not
get crushed. There are 2 methods of
hardening the tissues. One is by freezing them
and the other is by embedding them in a hard
material such at paraffin wax or gelatin.
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 Specimen…
 3. Smears: Smears are made from blood, bone
marrow or any fluid such as pleural or ascitic fluid.
These are immediately fixed in alcohol to presence
the cellular structures are then stained. Smears are
also made by crushing soft tissue between two
slides or an impression smear in made by pressing
a clean slide in contact with the moist surface of a
tissue. By doing this, the cells are imprinted on the
slide and these may be stained for cytological
examination.
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 Specimen…
• HISTOLOGY AND CYTOLOGY SPECIMEN:
 Specimen Labeling and Requisitions
Requirements :-
• Specimens will not be processed unless:
 1. All specimens labeled with: Patient Name,
DOB or MR#, Date and Time of collection,
Collector’s initials and Specimen Source .
 2. All requisitions adequately completed with
pertinent information regarding clinical history .
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 Specimen…
 3. Specimen label and requisition information match.
 4. Requisition is signed by the ordering physician
 TYPES OF HISTOLOGY SPECIMENS:
• Tissue specimens and biopsies ((e.g. placentas, renal,
liver, pleural biopsies, etc.)
 Collect in clean container with at least 10x the amount
of 10% Buffered Formalin per specimen size.

 Specimens must be completely covered and floating, for

adequate fixation.
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 Specimen…
 Lymph Node Studies and Frozen Sections
‘’SENT FRESH” Do Not Add Formalin to these
Specimens.
• KIDNEY BIOPSY :ALL Kidney biopsies MUST be
received “FRESH” on saline dampened gauze,
in a specimen container. The core sample
should be a minimum of 1mm in size with 3
mm being ideal for diagnostic interpretation.

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 Specimen…
 Bone Marrow Aspirations •
 BLADDER IRRIGATION (WASHINGS) :
• Obtain washings of the bladder with hormonal
saline or Ringer’s solution at the time of
cystoscopy.
 BREAST SMEARS and FINE NEEDLE
ASPIRATION FOR CYTOLOGY :

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 Specimen…
• Label two or more glass slides with patient’s
name and other information. After aspiration
has been completed, place a drop or two of the
material on the centers of as many glass slides
as possible, immediately covering each slide
with a second clean glass slide. After all slides
have been covered, gently pull the slides apart,
smearing the fluid along the length of each
slide. Fix slides IMMEDIATELY in 95% ethanol.
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 Specimen…
• The slightest air drying of the smear prior to
fixation will lead to some distortion of the cell
sample; therefore the smears that are fixed
immediately (wet fixed) will yield the best
preservation of the nuclear detail which is
mandatory for accurate interpretation.
• If there is any remaining material, draw saline
solution into the barrel of syringe, using the same
needle. Expel entire contents into specimen
container of CytoLyt Fixative (about 50cc)
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 Specimen…
 CEREBROSPINAL FLUID(CSF) :The majority of
specimens from the central nervous system
fluid compartment are drawn via lumbar
puncture. Cisternal and ventricular taps, as
well as direct aspirations of cystic or solid
masses also provide very useful cellular
samples. Knowledge of the collection method
and site is essential to the interpretation of
the cellular collection.
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 Specimen…
• Cellular deterioration is the main problem
encountered when delay occurs. Use second or
third sample drawn for cytology.
 GASTRO-INTESTINAL(GI) SPECIMENS :
 COLONIC BRUSHINGS : Brushings of mucosal
abnormalities at the time of fiber optic examination
of the colon are the generally accepted procedure
in patients suspected of harboring a colonic lesion.
•

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 Specimen…
• Specimen is smeared on clean glass slides and
fixed immediately by placing in 95% ethyl
alcohol.
• COLONIC WASHINGS :Preparation of the
patient is important since the colon must be
free of fecal material. The patient is instructed
to take 2 oz. of castor oil 12 hours prior to the
collection period and to have only a very light
meal on the morning of collection.
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 Specimen…
• Warm soap suds enemas are given until the
returns are clear. Two to three hours later the
specimen is collected by colonic irrigation.
Five hundred (500) ml. of warm (37º C.) saline
or Ringers solution is instilled with the patient
on the left decubitus position. Three to five (3-
5) minutes later the fluid is collected. Label
and send specimen to the Cytology Laboratory
accompanied by a Center ordered.
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 Specimen…
 DUODENAL and GASTRIC WASHINGS
• The patient should not have eaten food in the
previous eight hours
• The gastric tube should be passed to the 70 cm
mark without the use of any lubricants except
glycerin. Small sips of saline (not water) may be
used to aid in passing the tube. Two hundred fifty
(250) ml of Ringers solution or physiologic saline are
then introduced into the stomach in small
proportions, aspired and discarded.
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 Specimen…
• Introduce 250 ml of Ringers solution or saline
and re-aspirate and reintroduce the fluid
several times using a 50 or 100 ml. syringe and
with the patient in different positions (prone,
supine, left side, right side). The entire gastric
contents are then aspirated. Send specimen
to the Cytology Laboratory without delay,
accompanied by a Cerner ordered.

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 Specimen…
 ESOPHAGEAL WASHINGS :Material is best
obtained by direct esophagoscopy. Small
amounts of physiologic saline are injected
through the esophagoscope and aspirated.
Place the aspirate immediately in CytoLyt
fixative. Label and send to the Cytology
Laboratory immediately, accompanied by a
Cerner ordered.

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 Specimen…
 RECTAL WASHINGS :Preparation is the same as
for the Colonic Washings. Two to three hours
after the cleansing enema, 10 ml of normal
saline is introduced through the
sigmoidoscope and after 30 seconds is
aspirated by strong suction. Label and send to
the Cytology Laboratory accompanied by a
Cerner ordered:

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 Specimen…
 GYNECOLOGICAL SMEARS :Patient should not be
douched(vaginal rinsing) for 24 hours before
genital smears are obtained.
 •Preferably, smears should not be taken during
menstrual bleeding.
• No lubricant should be used.
• Requests for special testing (HPV; GC(Neisseria
gonorrhoeae). ChlamydiaTrichomonas(CT)must be
requested at time of Pap(papanicolaous) testing.
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 Review questions
• 1.What factors should be considered while you
are collecting histopathological specimen?
2.List different types of histopathological
specimens and how they are collected.

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 References
1. Sir William Lyon’s Road,John Crocker
the science of laboratory diagnosis, second edition
University of Warwick Science Park, Department of
Cellular Pathology,

• 2. Berhanu Seyum, Jemal Yimam Haramaya

university ,Histopathology for Medical Laboratory
Technology students in Ethiopia ,Lecture note
series in collaboration with The Carter Center ,2006
Addis Ababa.
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