Fluorescence Microscopy in Diagnostics and Immunology

based on ai / notes dobbelcheck.

Name some key figures in the development of fluorescence microscopy and their contribution year.

Köhler (1904), Haitinger (1938), Coons, Creech, and Jones (1941), Riggs et al. (1958), Leitz and Zeiss (1959), Tomlinson (1967).

List the advantages of fluorescence microscopy in medical diagnostics.

Fluorescence microscopy is: Easy, Sensitive, Specific, Rapid, Reliable, Universal, Adaptable.

What are some applications of fluorescence microscopy?

Infectious disease diagnosis, Autoimmune disease diagnosis, Environment monitoring, Emergency support, Biological research, Industrial process control, Disease diagnosis, Environment microorganism monitoring, Biological attack defense.

Define fluorescence.

Fluorescence is the ability of a fluorescent dye (fluorochrome) to emit radiation with a higher wavelength (emit light) than the exciting light after being irradiated with ultraviolet or blue-violet light.

What is a fluorochrome?

A fluorochrome is a chemical dye that fluoresces (emits light) when stimulated (excited) by certain wavelengths (colors) of light.

Why are immunofluorescence and fluorochroming important in diagnostics?

They are of utmost importance in diagnosing infectious diseases, especially bacterial infections.

What is fluorochroming (FC)?

Fluorochroming (FC) is frequently used in diagnosing bacterial infectious diseases, utilizing low concentrations of fluorochromes for staining bacterial pathogens.

Briefly outline Boya's method for detecting mycobacteria using fluorochroming.

1. Fixation: Fixate the slide with a solution mixture of auramin. 2. Rinse: Rinse with water. 3. Differentiation: Differentiate with acid alcohol. 4. Rinse: Rinse with distilled water. 5. Contrastive Staining: Stain the background with acid fuchsin solution. 6. Rinse and Dry: Rinse and dry the slide.

What is immunofluorescence (IM)?

Immunofluorescence uses marked antibodies of immune serum against an antigen to visualize it.

List fluorochromes used for marking antibodies in immunofluorescence.

Fluorescein isocyanate, Fluorescein isothiocyanate (FITC), Tetramethyl rhodamin isocyanate.

Describe the direct method of immunofluorescence.

In the direct method, binding occurs between the detected antigen and the marked antibody: Ag + Ab == AgAb.

Describe the indirect method of immunofluorescence (IMIF).

Immune serum (unmarked antibodies) serves as an interlayer against the detected antigen: Ag + Ab == AgAb; AgAb + Ab* == AgAbAb*.

What are the applications of immunofluorescence in laboratory diagnostics?

Detection of disease agents or their antigens; Detection of antibodies.

List the advantages of immunofluorescence microscopy.

Rapid results, Pure antigen not needed, Wide application, High sensitivity (detecting a few pathogens), Intracellular organisms are detectable, Simplicity (requires few steps), Low cost.

List the disadvantages of immunofluorescence microscopy.

Initial cost of the fluorescence microscopy equipment; Getting or training technical operators.