Name of Bright field Dark Field Fluorescence microscope Confocal laser Scanning Electron Transmission electron

microscope microscope (BFM) microscope microscope microscope (SEM) microscope (TEM)

(DFM)
Image

Principle The functioning of the When light hits an It uses a much higher A specimen is stained with The electrons are emitted The working principle of
microscope is based on object, rays are intensity light to illuminate fluorochrome is examined. after thermal energy is the Transmission Electron
its ability to produce a scattered in all the sample. This light excites When a beam of light is applied to the electron Microscope (TEM) is
high-resolution image azimuths or fluorescence species in the focused at a particular source and allowed to move similar to the light
from an adequately directions. The sample, which then emit light point of the fluoro- in a fast motion to the anode, microscope. The major
provided light source, design of the dark of a longer wavelength. The chromatic specimen, it which has a positive charge. difference is that light
focused on the image, field microscope is image produced is based on produces an illumination The beam of electrons microscopes use light rays
producing a high- such that it removes the second light source or the that is focused by the activates the emission of to focus and produce an
quality image. the dispersed light, emission wavelength of the objective lens to a plane primary scattered electrons image while the TEM uses
or zeroth order, so fluorescent species rather above the objectives. The at high energy levels and a beam of electrons to focus
that only the than from the light originally objective has an aperture secondary electrons at low- on the specimen, to produce
scattered beams hit used to illuminate, and excite, on the focal plane located energy levels from the an image.
the sample. the sample. above it, which primarily specimen surface.
functions to block any
stray light from reaching
the specimen.
Advantages - It is simple to use - Dark-field - This stems from its ability - It can be used to study - They are easy to operate - It produces very efficient,
with few adjustments microscopy to isolate individual proteins live and fixed cells and has user-friendly high-quality images with
involved while techniques are with a high degree of - It can be used to collect interfaces. high clarity.
viewing the image. almost entirely free specificity amidst non- serial optical sections - They are used in a variety - It can produce permanent
- It can be used to view of artifacts. fluorescing material. - It illuminates uniformly of industrial applications to images.
both stained and - A researcher can - The sensitivity is high across the focus points. analyze surfaces of solid - It is easy to train and use
unstained. achieve a dark field enough to detect as few as 50 objects. the Transmission Electron
by making molecules per cubic Microscope
modifications to micrometer.
his/her microscope.

Disadvantages - It has low contrast - The main - Fluorophores lose their - They have a limited - They are very expensive to - They are very big to
hence most specimens limitation of dark- ability to fluoresce as they are number of excitation purchase handle.
must be stained for field microscopy is illuminated in a process wavelengths, with very - They are bulky to carry - The preparation of
them to be visualized. the low light levels called photobleaching. narrow bands. - They must be used in specimens to be viewed
- Use of oil immersion seen response.in the Photobleaching occurs as the - They are expensive to rooms that are free of under the TEM is very
may distort the image final image. fluorescent molecules produce the ultraviolet rays vibrations and free of tedious.
- The use of coverslip - The sample must accumulate chemical damage used by the Confocal electromagnetic elements - The use of chemical
may damage the be very strongly from the electrons excited Microscopes fixations, dehydrators, and
specimen illuminated, which during fluorescence. - They are also expensive embedments can cause the
can cause damage - Cells are susceptible to to manufacture and to dangers of artifacts.
to the sample. phototoxicity, particularly purchase. - They are laborious to
with short-wavelength light. maintain.It requires a
constant inflow of voltage
to operate.
Applications - Used to visualize and - Useful for the - To identify structures in - Used to identify the - Used for spot chemical - To visualize and study cell
study the animal cells demonstration of fixed and live biological presence of fungal analysis in energy- structures of bacteria,
- Used to visualize and very thin bacteria samples. Fluorescence elements in the corneal Dispersive X-ray viruses, and fungi
study plant cells. not visible under microscopy is a common tool stroma, during Spectroscopy. - To view bacteria flagella
- Used to visualize and ordinary for today’s life science keratomycosis infection - Used in the analysis of and plasmids
study the illumination. research because it allows the and quick therapeutic cosmetic components which - To view the shapes and
morphologies of - Used for rapid use of multicolor staining, - Used in pharmaceutical are very tiny in size. sizes of microbial cell
bacterial cells demonstration labeling of structures within industries, to ensure the - Used to study the filament organelles
- Used to identify of Treponema cells, and the measurement of maintenance of thin-film structures of - To study and differentiate
parasitic protozoans pallidum in clinical the physiological state of a pharmaceuticals, microorganisms. between plant and animal
such as Paramecium. specimens. cell. cells.