--%>
Assignment Help - homework help

  Techniques of Immunofluorescence

Various Techniques of Immunofluorescence:

First utilized by Coon’s in the year 1942 for Pneumococcus in tissue sections.

Direct method: or single layer method

In this technique antigen is treated once with antibody tagged with fluorochrome.

1680_immunoflouroscence1.jpg


 Illustration: Explanation of Rabies antigen in impression smear.

1. Impression smear from hippocampus of infected dog is prepared and fixed.

455_immunoflouroscence2.jpg

2. This is treated with antirabies antibody tagged with Incubated and FITC.

1170_immunoflouroscence3.jpg

3. It is washed to eliminate un-reacted antibody.

4. The preparation is inspected for fluorescence.

318_immunoflouroscence4.jpg

Likewise different other antigens can be explained.

1. To explain viruses and bacteria in the respiratory specimens.
2. To explain Adenovirus, Chlamydia in the conjuntival smears.
3. To explain GC and Mycoplasma in urethral smear.

Indirect Technique or Antiglobulin Method or Double Layer Technique:

1667_immunoflouroscence5.jpg

Principle:

To explain the antigen, first unlabelled particular antibody is employed. Subsequently it is treated with tagged antibody to gamma globulin. (Therefore when the first layer is rabbit antibody then the second layer is anti-rabbit gamma globulin- tagged).

Procedure:

To explain an unknown antigen in fixed preparation. The test is completed in two steps:

Step: 1

1. Unknown antigen is existed in a fixed preparation.

837_immunoflouroscence6.jpg

2. It is treated with particular non-fluorescent serum.

1057_immunoflouroscence7.jpg

3. Incubated at 37ºC for 10 to 20 min in wet chamber.

711_immunoflouroscence8.jpg

4. Washed 3 to 4 successive baths for 5 min each in phosphate buffered saline. Antigen antibody complex is opposed to washings. The preparation is washed and then dried.

Step 2:

1. To the washed dried preparation labeled anti-species anti-globulin is added and incubated at 37ºC for 15 to 30 minutes in wet chamber. It is then washed and dried as in step 1. Whenever antigen-antibody complex has been made in the first step, labeled antiglobulin will couple to particular antibody to antigen and a finely fluorescent complex will be prepared.

 

Latest technology based Biology Online Tutoring Assistance

Tutors, at the www.tutorsglobe.com, take pledge to provide full satisfaction and assurance in Immunology help via online tutoring. Students are getting 100% satisfaction by online tutors across the globe. Here you can get homework help for Immunology, project ideas and tutorials. We provide email based Immunology help. You can join us to ask queries 24x7 with live, experienced and qualified online tutors specialized in Immunology. Through Online Tutoring, you would be able to complete your homework or assignments at your home. Tutors at the TutorsGlobe are committed to provide the best quality online tutoring assistance for Microbiology Homework help and assignment help services. They use their experience, as they have solved thousands of the Biology assignments, which may help you to solve your complex issues of Immunology. TutorsGlobe assure for the best quality compliance to your homework. Compromise with quality is not in our dictionary. If we feel that we are not able to provide the homework help as per the deadline or given instruction by the student, we refund the money of the student without any delay.