Types of microscopy

TYPES OF MICROSCOPE:
BASED ON APPLICATION:
Biological microscope Stereoscopic microscope
Magnification 4x to 100x Magnification 7x to 46x
Light comes beneath the stage Light comes from above and beneath the stage
Blood cells, bacteria and protozoa etc. Flower, rock, and insect etc.

BASED ON THE STRUCTURE:
Inverted microscope Upright microscope
Observe target from below Observe target from the above
Specimen observe on slides Specimen observe cells soaked with
culture in a dish.

Types of microscopy:
There are main types of microscopy which are as follows:
1- Optical microscopy or light microscopy
2- Electron microscopy
3- X-ray microscopy
4- Scanning probe microscopy

LIGHT MICROSCOPE:(1872) 200nm
Light microscope use the system of lens
Such as
Ocular lens
Objective lens
Light microscope used the to magnify the
image of smaller object to 100-1000 times.
Light beams travel between the Object and
eye to magnify the image.
Example:
RBC etc.

COMPOUND MICROSCOPE: 200nm
It is also called as bright field microscopy.
Principle:
A compound microscope uses a lens close
to the object being viewed to collect light
which focuses a real image of the object
inside the microscope.
That image is then magnified by a second
lens or group of lenses called the eyepiece
that gives the viewer an enlarged inverted
virtual image of the object.
Disadvantage:
Light dust easily observe.
e.g cheek cells views with bacteria

TYPES OF LIGHT MICROSCOPE:
1- ULTRA OR DARK FIELD MICROSCOPE:
Zsigmondy (1905)
It is used in microbiology and in autoradiography

Phase contrast microscope: (eraly1930)
A microscope that visualized minute surface by using light interference and observed the living
cell without staining.

DIFFERENTIAL INTERFERENCE CONTRAST
MICROSCOPY: (1952) Georges Nomarski
The microscope similar to phase contrast it observer the minute surface
irregularities but at a higher resolution

ULTRAVIOLET MICROSCOPE: Casperson (1938) Invented
ultraviolet microscope where the illuminating source is ultraviolet rays from mercury
or iodine-Quartz lamp. Image is captured on photographic film.

POLARIZING MICROSCOPE: (1768–1851)
Magnification power upto 2x to 100x.
PRINCIPLE:
A microscope that uses the different light transmission characteristics of materials.
Teeth, bone, striated muscle and gout crystal are evaluated by this microscope.

FLUORESCENCE MICROSCOPE: (1911-1913)
The "fluorescence microscope" refers to any microscope that uses fluorescence to
generate an image, fluorescence's such as fluoro chromes to get better resolution of the
fluorescent image. 8 October 2014 Nobel prize is given to Eric Betzig and his collagues
for super resolve fluorescence microscopy.

CONFOCAL MICROSCOPY:
Confocal microscope used the laser light for the image formation. The laser light scan the
sample that have been dyed. Theses samples are prepared on slides and inserted then with the
aid of dichromatic mirror the device produced a image on computer screen. Operated and
created a 3-D image. Like compound microscope this microscope offer a high degree of
magnification but resolution is more better. It is commonly used in cell biology and medicine
application

STEREOMICROSCOPY: (early 20th century) 120nm
Give the three dimensional view
an object. Magnification up to
300x. Use in medical, biological
and electronic industries.
A stereo model is an
optical microscope that functions
at a low magnification. It works
by using two separate optical
paths instead of just one. The two
objectives and two eyepieces
provide the eyes with slightly
different viewing angles.

ELECTRON MICROSCOPY: (1931-2000)
These microscope emit the electrons beams not light beams towards
target to magnify them.
TYPES OF ELECTRON MICROSCOPE:
1- Transmission electron microscope
2- Scanning electron microscope

SCANNING ELECTRON MICROSCOPE:
PRINCIPLE:
It is uses electron beams and sample
are scanned in vacuum or near
vacuum condition, after
dehydration sample is coated with a
thin film of conductive material
such as gold. After sample is
prepared placed in chamber than
SEM produces a 3-D image black
and white image on computer
screen.
It is used in physics, biology and
chemistry and examine the insect to
bone through SEM.

TRANSMISSION ELECTRON MICROSCOPE:
PRINCIPLE:
Transmission electron
microscopy (TEM) is
a microscopy technique in which a
beam of electrons is transmitted
through an ultra-thin specimen,
interacting with the specimen as it
passes through it.
It works on the principle of nature
of electrons as they exhibit less
wavelength when bombarded at
high velocity.
It is used see the 2-D image of
object.

SCANNING PROBE MICROSCOPY: (1981)
PRINCIPLE:
Scanning Probe Microscopy ( SPM )
scans an atomically sharp probe over
a surface, typically at a distance of a
few nanometres.
The interaction between the sharp
probe and surface provides 3D
topographic image of surface at the
atomic scale. SPMs are relatively
simple, inexpensive, and easy to
operate.
ATM and SNOM

X-RAY MICROSCOPY:
PRINCIPLE:
An X-ray microscope uses electromagnetic radiation in the soft X-ray band to produce
magnified images of objects. Therefore, an X-ray microscope exposes film or uses a charge-
coupled device detector to detect X-rays that pass through the specimen. The resoultion power
of x-rays lie in between the optical and electron microscopy. X-rays do not reflect or refract
easily, and they are invisible to the human eye.

OPTICAL
MICROSCOPY
Resolution to
approximately
0.2 micrometres
This limits the
practical magnification
limit to ~1500x.
Out-of-focus light from
points outside the focal
plane reduces image
clarity.
Internal structure
difficult to observe.
ELECTRON
MICROSCOPY
 Large
 Expensive
piece of
equipment
extremely
sensitive to
vibration and
external
magnetic fields.
Scanning probe
microscopy
Slower in
acquiring
the image.
Maximum
image size is
smaller.
X-rays
microscope
 Due to the high
energy destroyed
the sample and
cause the damage
of cells.
 Mutations
DISADVANTAGE

Recent development:
Howard Hughes Medical Institute's Janelia Research Campus, and colleagues report
the work April 19, 2018.
Before betzig,s and his colleagues work combining the two microscope techniques
together in 2014.
Lattice light sheet microscope
Two adaptive optical system to maintain the thin illumination of a lattice light sheet as
it penetrates within an organism, to create distortion-free images when looking down
on the illuminated plane from above. By shining a laser through either pathway, the
researchers create a bright point of light within the region they wish to image.
Movement of cells.
LIMITATIONS:
Lattice light sheet microscopy is limited to transparent samples and thin samples to
achieve good image quality

Types of microscopy

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Types of microscopy