MICROSCOPY

By: Alyanna Angel Carman

Anne Valerie Lasaleta
Kate Samel Panilla
MICROSCOPY

 Microscopy is the technical field of using microscopes to

view objects and areas of objects that cannot be seen with the
naked eye (objects that are not within the resolution range of
the normal eye).
MICROSCOPE
 The microscope is a valuable instrument. There are many
small objects or details of objects which cannot be seen by the
unaided human eye. The microscope magnifies the image of
such objects thus making them visible to the human eye.
Microscopes are used to observe the shape of bacteria, fungi,
parasites and host cells in various stained and unstained
preparations.
 TYPES OF MICROSCOPY

 Microscopes used in clinical practice are

light microscopes.
 A compound light microscope is the most common microscope
used in microbiology. It consists of two lens systems
(combination of lenses) to magnify the image.
 Each lens has a different magnifying power. A compound light
microscope with a single eye-piece is called monocular; one
with two eye-pieces is said to be binocular.
 TYPES OF MICROSCOPY

 Microscopes that use a beam of electrons (instead of a

beam of light) and electromagnets (instead of glass
lenses) for focusing are called electron microscopes.
 LIGHT MICROSCOPY
Brightfield microscopy
 This is the commonly used type of microscope. In
brightfield microscopy the field of view is brightly lit so
that organisms and other structures are visible against it
because of their different densities. It is mainly used with
stained preparations.
 LIGHT MICROSCOPY

Darkfield microscopy
 In darkfield microscopy the field of view is dark and the
organisms are illuminated. A special condenser is used
which causes light to reflect from the specimen at an
angle.
 LIGHT MICROSCOPY
Phase-contrast microscopy
 Phase-contrast microscopy allows the examination of
live unstained organisms. For phase-contrast microscopy,
special condensers and objectives are used. These alter
the phase relationships of the light passing through the
object and that passing around it.
 LIGHT MICROSCOPY

Fluorescence microscopy
 In fluorescence microscopy specimens are stained with
fluorochromes/ fluorochrome complexes. Light of high
energy or short wavelengths (from halogen lamps or
mercury vapour lamps) is then used to excite molecules
within the specimen or dye molecules attached to it.
 PARTS OF THE MICROSCOPE

 The main parts of the microscope are the eye-pieces,

microscope tube, nosepiece, objective, mechanical stage,
condenser, coarse and fine focusing knobs, and light
source.
 PARTS OF THE MICROSCOPE
 Eyepiece: The lens the viewer looks through to see the
specimen. The eyepiece usually contains a 10X or 15X
power lens.

 Diopter Adjustment: Useful as a means to change focus

on one eyepiece so as to correct for any difference in
vision between your two eyes.
 PARTS OF THE MICROSCOPE
 Body tube (Head): The body tube connects the eyepiece
to the objective lenses.
 Arm: The arm connects the body tube to the base of the
microscope.
 Coarse adjustment: Brings the specimen into general
focus.
 PARTS OF THE MICROSCOPE
 Fine adjustment: Fine tunes the focus and increases the
detail of the specimen.

 Nosepiece: A rotating turret that houses the objective

lenses. The viewer spins the nosepiece to select different
objective lenses.
 PARTS OF THE MICROSCOPE
 Objective lenses: One of the most important parts of a
compound microscope, as they are the lenses closest to the
specimen.
 Specimen or slide: The specimen is the object being examined.
Most specimens are mounted on slides, flat rectangles of thin
glass.
 Stage: The flat platform where the slide is placed.
 PARTS OF THE MICROSCOPE
 Stage clips: Metal clips that hold the slide in place.
 Stage height adjustment (Stage Control): These knobs move
the stage left and right or up and down.
 Aperture: The hole in the middle of the stage that allows light
from the illuminator to reach the specimen.
 On/off switch: This switch on the base of the microscope turns
the illuminator off and on.
 PARTS OF THE MICROSCOPE

 Illumination: The light source for a microscope. Older

microscopes used mirrors to reflect light from an external
source up through the bottom of the stage; however, most
microscopes now use a low-voltage bulb.
 Iris diaphragm: Adjusts the amount of light that reaches the
specimen.
 PARTS OF THE MICROSCOPE

 Condenser: Gathers and focuses light from the illuminator onto

the specimen being viewed.

 Base: The base supports the microscope and it’s where

illuminator is located.
 HOW DOES A MICROSCOPE
WORK?
 All of the parts of a microscope work together - The light from
the illuminator passes through the aperture, through the slide,
and through the objective lens, where the image of the specimen
is magnified.
 The then magnified image continues up through the body tube
of the microscope to the eyepiece, which further magnifies the
image the viewer then sees.
 INSTALLATION AND
STORAGE

 Install the microscope on a sturdy, level table. Equipment and

instruments which generate vibrations, such as centrifuges and
refrigerators, should not be placed on or near this table.
 The height of the table should be convenient for the user. As an
alternative or in addition, an adjustable stool should be made
available to make microscopy comfortable.
 INSTALLATION AND
STORAGE

 The table should be away from water, sinks, and racks

containing chemicals, to prevent damage to the microscope
from splashes or spills.
 If the microscope does not have a built-in light source then the
table should be placed near a window away from direct sunlight
and arrangements made for the provision of a lamp.
 MAINTENANCE OF LENSE

 Avoid collection of dust and immersion oil on the objectives

and eye-pieces by keeping the microscope covered. Do not
allow immersion oil to touch any of the objectives other than the
oil immersion objective. Always keep the eye-pieces in place to
protect the inner surface of the objective. Close the holes of
missing objectives in the nose-piece by using special caps that
are provided, or by sealing with adhesive tape.
 REMOVAL OF DUST FROM LENSES

 Removal of dust from lenses Check for dust or dirt on

the lenses (eye-pieces, objective, condenser and
illuminator lenses) if the image appears hazy or with
black dots.
 STIFFNESS

 is due to accumulation of dust or because the sliding channel

has become rough. This problem can be overcome by cleaning,
polishing and lubricating the sliding channel and the rack and
pinion
 Stiff movements may also be due to mechanical bending of
some part. Rectify the fault or call the service engineer.
TISSUE
CULTURE
 TISSUE CULTURE

 Tissue culture is the growth of tissues or cells in an artificial

medium separate from the organism. This is typically facilitated
via use of a liquid, semi-solid, or solid growth medium, such as
broth or agar.
 TISSUE CULTURE

 A whole plant can be regenerated from a small tissue or plant

cells in a suitable culture medium under controlled environment.
The plantlets so produced are called tissue-culture raised plants.
 This capacity of a single cell to grow into a complete plant is
termed as Totipotency.
 TISSUE CULTURE PROCESS

 STAGE I is the initiation phase. It concerns the establishment of plant

tissue in vitro by sterilizing the material and initiating it into culture.
 STAGE II is the multiplication phase. At this stage, the in vitro plant
material is re-divided and placed in a medium with plant growth
regulators that induce the proliferation of multiple shoots.
 STAGE III is the root formation phase. It involves the introduction of
hormones to induce rooting and the formation of complete plantlets.
Flasks containing tissue culture 
growth medium which provides
nourishment to growing cells
 ADVANTAGES AND BENEFITS

 are guaranteed to be disease free

 have a more fibrous, healthier root system free of any root rot problems
 exhibit a denser, bushier branching habit
 are characterised by more vigorous growth after transplanting
 have a higher survival rate
 are ready for re-sale in a shorter time
END