PRINCIPLES AND FEATURES OF LIGHT

MICROSCOPES
 PRINCIPLES OF MICROSCOPY

0.1 mm or 10th of a millimeter

Human Egg cell

Eukaryotic
Bacterial
cell size
cell size
1-20 μm
0.1-5 μm
MAGNIFICATION AND RESOLUTION

De
ana tailed re r
lys e a
is Cl

With increased magnification

and resolution pictures are
clearer and also more detail
can be seen.
 MAGNIFICATION AND RESOLUTION

Clarity &
Resolution Sharpness
change of the image.

More Resolution Less Resolution

 MAGNIFICATION AND RESOLUTION

R Seen as one object

es
The two objects can be ol
seen as overlapping / not ut
clear if the resolution of io
n
the microscope is not
great or its not a in
powerful microscope. cr
ea
se Seen as two separate objects
MAGNIFICATION AND RESOLUTION
MAGNIFICATION AND RESOLUTION

Resolution adjustment
THE LIGHT MICROSCOPE
THE LIGHT MICROSCOPE

The light microscopes do not require cells

to be killed first for viewing purpose.
Thus, many cells can be seen in real time
performing functions.

Cells have
3-D shapes
 THE LIGHT MICROSCOPE

Visible light range

A typical human eye will

respond to wavelengths
from about 380 to 740
nanometers.
Electromagnetic Spectrum

The 200nm resolution means, if we want to see
two objects at a distance of 200nm, then it can
only be seen at 200nm apart distance. Lesser than
this will make them appear overlapping / blur /
not clearly distinct from each other.
Specimen compared to the image
is 2000 times smaller, and image is
2000 times bigger than the real
specimen.
SIZE UNIT
SIZE UNIT
 SIZE UNIT

Ant small plants animal cell Bacteria Mitochondrion Viruses fibrils molecules

mm μm nm
 SIZE UNIT

Example:
m 1m = 100cm = 1000mm
×1000

÷ 1000 mm
×1000 1m = 1000,000 μm

÷ 1000
μm
SIZE UNIT

0.04m × 1000,000 = 40,000 μm

 M A G N I F I C AT I O N C A L C U L AT I O N S

Calculation of Magnification of an object.

I = (to be measured from Question paper by a ruler)
A = Actual size (image size given in question paper)
m = Magnification

Magnification = I / A

Actual size = image size / magnification

ELECTRON MICROSCOPE
 ELECTRON MICROSCOPE

Visible Light Source Electron Beam

Beam
Detector

Light Microscope Electron Microscope Specimen

 ELECTRON MICROSCOPE

Wavelength of visible light is longer than electron beam.

Light Microscope Electron Microscope

ELECTRON MICROSCOPE

Beam of Electrons

Magnets

Electron
detector

Specimen
ELECTRON MICROSCOPE
TRANSMISSION AND SCANNING
MICROSCOPY
 SCANNING ELECTRON MICROSCOPE

Electron

Specimen is coated with

metal and placed in vacuum Magnets

Electron detector

Specimen

Electrons scattered
SCANNING ELECTRON MICROSCOPE

When the electrons scatter

they form 3-D image.
Resolution and magnification
of the scanning electron
microscope is low.
 TRANSMISSION ELECTRON MICROSCOPE

Electron

Specimen is coated with

metal and placed in vacuum Magnets

Specimen

Electron detector
Present underneath
TRANSMISSION ELECTRON MICROSCOPE

In transmission electron
microscope the image
formed is 2-D image.
Resolution and magnification
of the transmission electron
microscope is high.
SUMMARY OF TEM AND SEM

Lower the nm no. higher the resolution

SUMMARY OF DIFFERENT MICROSCOPES

In general for E.M. 0.5 – 1 nm is acceptable.