A fluorescence microscope uses fluorescence to enhance its capabilities beyond a regular light microscope. It illuminates samples tagged with fluorescent dyes with high-energy light, which causes the dyes to emit lower-energy light, producing a magnified image. This allows visualization of cell structures and live/dead cell assays. Advanced fluorescence microscopes like confocal microscopes can generate high-resolution 3D images of sample depths using lasers and image reconstruction software. Key applications include imaging cellular components, viability studies, and fluorescence in situ hybridization.

1. Fluorescence microscope
Presented by : - Salvi Pinky P.
M.sc sem : - 2
CBO : - 406
Department of Life sciences
H. N. G. U., Patan

2. Content
 What is fluorescence microscope ?
Principle
Application

3. What is fluorescence microscope ?
• Fluorescent is a type of a light microscope.
• Fluorescent microscope is much the same as a Conventional light
microscope with added features to enhance its capabilities.
• This fluorescent species in turn emits a lower energy light of a
longer wavelength that produces the magnified image instead of
the origin light source.

4. • It is also used to visually enhance 3 - D features at small scales.
• This is achieved by using powerful light sources, such as lasers,
that can be focused to a pinpoint.
• This focusing is done repeatedly throughout one level of a
specimen after another.
• Most often an image reconstruction program pieces the multi
level image data together into a 3 - D reconstruction of the
targeted sample.

6. Principles
• In most cases the sample of interest is labeled with a
fluorescent substance known as a fluorophore and then
illuminated through the lens with the higher energy source.
• The illumination light is absorbed by the fluorescence (
now attached to the sample ) and causes them to emit a
longer lower energy wavelength light.
• This fluorescent light can be separated from the
surrounding radiation with filters designed for that specific
wavelength allowing the viewer to see only that which is
fluorescing.

7. • The basic task of the fluorescence microscope is to let excitation
light radiate the specimen and then sort cut the much weaker
emitted light form the image.
• First, the microscope has a filter that only lets through radiation
with the specific wavelength that matches your fluorescing
material.
• The radiation collides with the atoms in your specimen and
electrons are excited to a higher energy level.
• When they relax to a lower level, they emit light.

9. • To become detectable ( visible to the human eye ) the
fluorescence emitted from the sample is separated from the
much brighter excitation light in a second filter.
• This works because the emitted light is of lower energy and has
a longer wavelength than the light that is used for illumination.
• Most of the fluorescence microscope used in biology today are
epi - fluorescence microscopes, meaning that both the excitation
and the observation of the fluorescence occur above the sample.
• Most use a xenon or Mercury arc - discharge lamp for the more
intest light source.

10. Application
• The refinement of epi - fluorescent microscopes and advent of
more powerful focused light sources, such as lasers, has led to
more technically advanced scopes such as the confocal laser
scanning microscopes and total internal reflection fluorescence
microscopes ( TIRF ).
• CLSM'S are invaluable tools for producing high resolution 3 - D
images of subsurfaces in specimens such as microbes.
• Their advantage is thick samples at various depths by tacking
images point by point and reconstructing them with a computer
rather than viewing whole images through an eyepiece.

11.  These microscopes are often used for : -
• Imaging structural components of small specimens, such as
cells.
• Conducting viability studies on cell populations ( are they a live
or dead ? ).
• Imaging the genetic material within a cell ( DNA & RNA ) .
• Viewing specific cells within a larger population with
techniques such as FISH.

12. References
 https://www.directindustry.com
 https://www.en.m.wikipedia.org
 https://www.biotek.com
 https:// www.serc.carleton.edu

13. THANK YOU