MICROSCOPY

Prelims
BSMT – 2B GIESSAN MAE LABRADO

MICROSCOPE COMPONENTS OF MICROSCOPE

MECHANICAL SYSTEM

BODY
• Base - It attaches the tube with the
rest of microscope
• Arm - It provides support to the body and tube
at correct height
• Revolving nosepiece - holds two or more
objective lenses and can be rotated to easily
change power revolving nosepiece

• The word microscope comes from a Greek MECHANICAL STAGE

word “micros” meaning “small” and “skopein” • Vernier scale - used to trace the part of the
meaning “to look at” or “to view”. This was blood film you need to reexamine.
coined by Giovani Farber in 1628. • Feed knob/Stage control - to navigate the x
• A device used in laboratory to observe and y axis
objects which cannot viewed with a naked
eye. OBSERVATION TUBE/HEAD
• Diopter - used to compensate the difference
PERSONS WHO CONTRIBUTED TO THE in eyesight between the left and right eye
DEVELOPMENT OOOF THE MICROSCOPES • Interpupillary Distance - used to adjust the
eyepieces in relation to the distance between
Hans and Zacharias Jansen – Devised the first the left and the right eye
microscope in 1950
Robert Hooke – First to use the compound ADJUSTMENT SYSTEM
microscope in 1554. Hooke studied the great
diversity of material from household objects, plants COARSE ADJUSTMENT KNOB
and trees. • Located on either side of the arm. Moves the
o He described spots of mold he found (hairy stage to bring the object into focus
mold) on the sheepskin cover of a book
FINE ADJUSTMENT KNOB.
Anton Van Leeuwenhoek - First microscopist • Located within the coarse adjustment knob.
o Father of microbiologist Allows fine focusing of the specimen.
o He described the bacteria and protozoan
which called “animalcules” OPTICAL SYSTEM

Christian Hyugens – invented the simple but OBJECTIVE LENSES

effective two lens eyepiece in 1684 • Located on the revolving nosepiece. Each
Charles Spencer – designed advance microscope lens has a different magnifying power.
objectives by introducing higher numerical aperture o The smallest objective (scanning
Francis Wenham – Invented the Darkfeild objective) is the smallest magnification at
microscope 4x
Fritz Zernicke – invented the Phase Contrast o Low power objective at 10x
microscope in 1935 o High power objective at 40x
E.N Harvey – invented the Centrifuge microscope o Highest magnification (immersion oil
Knoll et.Al – invented the Electron microscope objective) at 100x.
MICROSCOPY
Prelims
BSMT – 2B GIESSAN MAE LABRADO

OCULAR (EYEPIECE) PHASE CONTRAST MICROSCOPE

➢ The lens in the upper part of the microscope.
Monocular microscopes have one ocular, Phase-contrast
while binocular microscopes have two microscopy visualizes
oculars. Ocular magnification is 10x differences in the refractive
REVOLVING NOSEPIECE indexes of different parts of a
➢ Located at the lower end of the body tube. A specimen relative to unaltered
revolving device that holds the objective light. It allows for the detailed
lenses (aka objectives) observation of living organisms,
especially the internal
ILLUMINATION SYSTEM structures

CONDENSER
➢ Lens beneath the stage that concentrates and • This microscope splits a beam of light into 2
focuses the light before it passes through the types of light, direct and refracted (reflected)
specimen to be viewed and brings them together to form an image of
IRIS DIAPHRAGM LEVER the specimen.
➢ Small lever beneath the condenser which • Where the lights are “in-phase” the image is
controls the amount of light passing through brighter, where the lights are “out of phase”
the condenser. the image is darker, and by amplifying these
LIGHT SOURCE differences in the light, it enhances contrast
➢ Provides a beam of visible light to be passed
through the specimen. DARKFIELD MICROSCOPE

Magnification – the increase of an object’s apparent Dark-field microscopy is

size. ideally used to illuminate
Resolution - The power to show details clearly unstained samples causing
them to appear brightly lit
against a dark background.
It shows a light silhouette of
an organism against a dark
background.

In dark-field
microscopy, the light
TYPES OF COMPOUND MICROSCOPE
reaches the specimen
from an angle with the
help of an opaque disk.
LIGHT MICROSCOPY

An instrument that
magnifies an image and
allows visualization of The specimen
greater detail than is appears lit up
possible with the unaided against a dark
eye background
MICROSCOPY
Prelims
BSMT – 2B GIESSAN MAE LABRADO

FLUORESCENCE MICROSCOPE

Fluorescence microscopy
is used to study specimens
that are chemically
manipulated to emit light. It
is currently one of the more
powerful and versatile The light source in a
techniques available for confocal microscope comes from
studies of biologic an illuminating laser light system
specimens. that is strongly convergent and
therefore produces a high-intensity
excitation light in the form of a
In fluorescence microscopy, specimens are shallow scanning spot.
first stained with fluorochromes and then viewed
through a compound microscope by using an ELECTRON MICROSCOPE
ultraviolet (or near-ultraviolet) light source.
Electron microscopy uses magnetic coils to
direct a beam of electrons from a tungsten filament
Microorganisms through a specimen and onto a monitor.
appear as bright
objects against a TWO KINDS OF EM
dark background 1. Transmission electron microscope (TEM)
• It transmits electrons through an ultra-thin
specimen to reveal its internal structure in
2D with very high magnification and
Fluorescence microscopy is used primarily in resolutions uses the interaction of 8 beam
a procedure called fluorescent- antibody (FA) of electrons with 8 specimens to produce
technique, or immunofluorescence. an image.

CONFOCAL SCANNING MICROSCOPE 2. Transmission electron microscope (TEM)

• scans the surface of a bulk sample with
an electron beam, detecting emitted
electrons to create a 3D-like image of the
surface topography. The electron beam
does not pass through the specimen but
is scanned across its surface.

ATOMIC FORCE MICROSCOPY

Atomic Force Microscopy (AFM) takes a
fundamentally different approach by "feeling" the
This allows visualization of a biologic surface with a sharp tip on a cantilever, measuring
specimen in three dimensions. The two lenses in the the forces between the tip and the sample to create a
confocal microscope (objective and photo· tube lens) 3D topographical map at the nanoscale.
are perfectly aligned to focus light from the focal point
of one lens to the focal point of the other lens
MICROSCOPY
Prelims
BSMT – 2B GIESSAN MAE LABRADO

CARE AND MAINTENANCE

DONT’s

1. Always carry the microscope in an upright position ➢ Clean the lenses of the objectives and
using both hand sone hand at the base and the other eyepieces with Xylene
at the arm. ➢ Use ordinary paper or cotton wool to clean
2. Keep the microscope away from the edge of the the lenses
table, and do not let the cord get entwined around or ➢ Touch the objectives with your finger
with something ➢ Clean the inside lenses of the eyepiece and
3. When returning the microscope to its proper place, the objectives with cloth or paper Leave the
ALWAYS: microscope without the eyepiece
• Remove the slide from the mechanical ➢ Carry the microscope by the limb with one
stage hand
• Rotate the nosepiece so that the scanning ➢ Exchange lenses from different microscope
lens is in place. ➢ Dismantle or try to clean parts of the
• Secure the cord so that it does not hang microscope difficult to reach, unless you have
down. been trained to do so.
• Make sure that the equipment is turned or ➢ Subject the microscope to severe and sudden
switched off. impact
• Cover it with dust jacket when not in use ➢ Touch the bulb of the microscope with bare
fingers D
4.Always clean the microscope. Materials for
Cleaning:
• Piece of cotton cloth and lens paper •
70% Alcohol
• Special Tissue Paper or Whole
Absorbent paper
• Plastic Cover
• Make-up Brush (Softest )
• Rubber Bulb

DO’s

➢ Cover the microscope when not in use.

➢ Line up the x10 objective with the ocular
when microscope is not in use
➢ Report to the Maintenance of any
malfunctioning observed
➢ Oil immersion objective should be cleansed
after
➢ Always put the microscope on a flat and
sturdy surface
➢ Keep the microscope in a dust-free area
➢ Always put the grounding wire in place.