Fluorescence and Confocal Microscopy Outline

LovedBinomial avatar
LovedBinomial
·
·
Download

Start Quiz

Study Flashcards

29 Questions

What type of chemicals are fluorochromes?

Photoreactive chemicals

What happens to the light absorbed by fluorochromes?

It is emitted at a lower energy level as fluorescence

What is the primary function of the secondary antibody conjugated to a fluorochrome in immunofluorescent staining?

To allow detection of antigen-antibody complexes

What type of tissues is immunofluorescent staining suitable for?

Frozen, non-fixed tissues and ethanol fixed tissues

What is the main purpose of immunofluorescent staining?

To identify patterns of protein expression in cells

What is the role of fluorophores in fluorescence microscopy?

To absorb light and emit it with lower energy

What is the purpose of incubating cells with a blocking solution in the immunofluorescent cell staining process?

To minimize non-specific staining

Which fluorochrome is the secondary antibody directed against the primary antibody in the provided information?

FITC

What is used to visualize stained specimens on a fluorescent microscope?

PBS/Glycerol mix

What is the major problem in microscopic examination of fluorescent specimens?

Photobleaching

Which chemical agent may be added to the mounting medium to preserve specimen brightness and prevent photobleaching?

Trans-pyridine-2-azo-p-dimethylaniline (PADA)

What is the purpose of incubating fixed cells in 1% Triton X-100 in PBS+0.02%BSA for 2 minutes at room temperature?

To permeabilize the cells

Which primary antibody is specific for the protein p53 as mentioned in the provided information?

Sheep anti-p53 polyconal

What is the primary purpose of immunofluorescent staining?

To locate and identify patterns of protein expression in cells.

What is the function of fluorochromes in fluorescence microscopy?

Fluorochromes absorb light in a specific wavelength range and re-emit it with lower energy, allowing the visualization of fluorescent samples.

What type of tissues is immunofluorescent staining suitable for?

Frozen, non-fixed tissues and ethanol fixed tissues.

What happens to the light absorbed by fluorochromes?

Fluorochromes re-emit the absorbed light at a longer wavelength with lower energy, a process known as fluorescence.

What is the major advantage of confocal microscopy over conventional fluorescent microscopy?

Confocal microscopy provides higher resolution and better 3-dimensional imaging compared to conventional fluorescent microscopy.

What is the role of the secondary antibody conjugated to a fluorochrome in immunofluorescent staining?

To bind to the antibody-antigen complex and allow detection of the complex through fluorescence.

What chemical can be added to the mounting medium to preserve specimen brightness and prevent photobleaching?

trans-pyridine-2-azo-p-dimethylaniline (PADA)

What is the specific fluorochrome used for the secondary antibody directed against the primary antibody in the provided information?

FITC

In immunofluorescent cell staining, what is the primary function of the secondary antibody conjugated to a fluorochrome?

To bind to the primary antibody and aid in visualization under a microscope.

What is the role of fluorochromes in fluorescence microscopy?

Fluorochromes absorb light at one wavelength and emit it at a longer wavelength, producing fluorescence, which aids in visualization under the microscope.

What chemical is used to fix cells during the cell preparation process?

Paraformaldehyde or methanol/acetone

What is the major problem in microscopic examination of fluorescent specimens?

The tendency of fluorochromes to lose fluorescence upon excitation by a high energy light source, known as photobleaching.

What is the purpose of incubating fixed cells in 1% Triton X-100 in PBS+0.02%BSA for 2 minutes at room temperature?

To permeabilize the cells, allowing antibodies to enter and bind to intracellular structures or proteins.

What is used to visualize stained specimens on a fluorescent microscope?

Mounting medium, usually a PBS/Glycerol mix

What is the specific primary antibody mentioned for the protein p53 in the provided information?

Sheep anti-p53 polyconal

What is the specific fluorochrome used for the secondary antibody directed against the protein a tubulin in the provided information?

Texas Red

Learn about immunofluorescent staining, conventional fluorescent microscopy, confocal microscopy, and their applications in this presentation by Dr. Firas Zakaria and Dr. Hassan Khachfe. Understand the use of fluorochromes as fluorescent markers in microscopy and the specific requirements for suitable samples.

Make Your Own Quizzes and Flashcards

Convert your notes into interactive study material.

Get started for free
Use Quizgecko on...
Browser
Browser