Immunofluorescence

Question 1

What are the advantages of using Fluorescence microscopy:

Answer 1

Easy, sensitive, specific, rapid, reliable, universal and adaptable

Question 2

What is the principle of fluorescence:

Answer 2

• The ability of fluorescent dye (fluorochrom) to emit the radiation with higher wavelength (emit light) than was exciting light after irradiation with ultraviolet or blue-violet light
• wavelength of emitted radiation is usually green, greenish-yellow or orange

Question 3

Fluorochroming (FC) advantage and used for:

Answer 3

Advantage
* High contrast of colour transformation
* Possibility of fast examination
* High susceptibility

Method for visualization of bacteria by FC
* Boya’s method for detecton of Mycobacteria (Tuberculous germs are stained yellow orange at dark nonfluorescing background

Question 4

What is the principle of Immunofluorescence (IM):

Answer 4

Using marked antibodies of immune serum against antigen which we want to visualise

Question 5

A conjugate is a:

Answer 5

Marked antibody

Question 6

What does conjugation mean:

Answer 6

Chemical reaction between immunoglobuline of serum and fluorochrome

Question 7

Direct method of immunofluorescence:

Answer 7

Binding between detected antigen and marked antibody

Question 8

Indirect method of immunofluorescence:

Answer 8

Can be used for detection of antigens and also for detection of serum antibodies

Question 9

Advantages of Immunofluorescence microscopy:

Answer 9

Rapid results, pure antigen not needed, wide application, high sensitivity, intracellular organisms, simplicity and low cost

Question 10

What are the disadvantages of Immunofluorescence microscopy:

Answer 10

• Initial cost of the equipment
• Getting or training technical operators