Académique Documents
Professionnel Documents
Culture Documents
Compiled by
Dr.CH.Aruna kumari PG
Guided by
Dr. K.R.L Surya Kirani MD Prof&HOD
Dr. K. Rajasekhar MD Asst.Prof
Dr. S.S.Sarma, MD Asst.Prof
History
The term microscope was coined by Faber in 1623.
MICROSCOPY :
Applications:
2.Compound microscope.
1.Simple microscope
Light Electron
Microscopes Microscopes
Phase Transmission
Bright Field Dark Field Flourescent Scanning EM
Contrast EM
COMPOUND MICROSCOPE
• This is an optical instrument containing one or
more lenses producing an enlarged image
WORKING OF COMPOUND MICROSCOPE
1. Magnification
2. Resolution
3. Contrast
Magnification:
• Field lens collect them to pass through eye lens that magnifies it
further.
• NA = n Sin θ
n = refractive index,
θ = angle of aperture
Examples :
Achieved by staining.
Uses
PRINCIPLE
For detecting:
HISTORY
Release
Absorb
light of
Molecules radiant Excited
longer
energy
wavelength
Tissues,cells,bacteria
Stained with
fluoroscent dye
Examined under
microscope with UV
light
Luminous objects
around dark
background
FLUOROCHROMES
FADING
Reduction of intensity of fluorescence.
Principle:
Increased sensitivity.
1. To measure
Redox potentials
Fluidity of membranes
• Non hazardous.
LED FM ZN
3+ 3+
2+ 2+
1+ 1+
Negative Negative
PHASE CONTRAST MICROSCOPE
PHASE PLATE
Placed in back focal plane of objective.
FUNCTION :
TRICHOMONAS ENTAMOEBA
Differential Interference Contrast Microscopy
PRINCIPLE
1. ELECTRON GUN
2. ELECTROMAGNE
TIC LENSES
3. VACUUM PUMPS
4. OPENING TO
INSERT
SAMPLES
5. OPERATION
PANEL
6. DISPLAY
SCREEN
7. WATER SUPPLY
TO COOL THE
INSTRUMENT
Transmission electron microscope
Specimen Preparation
1) NEGATIVE STAINING
5) Microtomy
Advantages
1) No 3-D image.
H.pylori Rotavirus
Scanning electron microscope
Secondary e-
Collector
Detector—emits light
when struck by e-.
Light converted to
current.
Cathode ray tube
screen
Specimen preparation
Easy.
Avian flu
RBC
NEURON
ADVANTAGES OF SEM
Advantages :
5kV Low-voltage EM
1. High contrast
Problems
• Formation of artefacts
Comparision of various types of Microscopes
light vibrates in
all planes that contain
vibration
the light ray
direction
(i.e., all planes
perpendicular to
the propagation
direction)
Polarized Light
Unpolarized Light
• Detects specimens that are birefringent(have the
characteristic of double refraction, i.e. the velocity of
light refracted by a substance is not the same in all
directions).
Compared to a typical microscope,a polarizing microscope
has the following added units:
4) An analyzer
-crystals
-minerals
-certain foreign bodies and
-some lipids.
Disadvantages
2. Microdissection
3. Examination of parasites
3. Flow cytometry
CONFOCAL MICROSCOPE
USES
Types :
Principle:
1. Localization microscopy
2. Localization microscopy SPDM
3. SPDM Phymod
4. STORM – Stochastic optical reconstruction
microscopy
PALM - Photoactivated localization microscopy
FPALM - Fluorescence photo activation localization
microscopy
5. Label- free localization microscopy
Combination of techniques: