immunfluoresence
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homeThis document provides an overview of immunofluorescence testing. It defines immunofluorescence as a process using fluorescent dyes and antibodies exposed to ultraviolet light to detect antigens and antibodies. There are two main types: direct immunofluorescence detects unknown antigens using a single layer of fluorescent antibody, while indirect immunofluorescence detects antibodies by first forming an antigen-antibody complex and then adding a fluorescent secondary antibody. Examples of applications include detecting viruses, bacteria, and toxoplasma antibodies. The procedure and results of both direct and indirect immunofluorescence are described.
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_ู
ุงูู_ุฏุฑุฎ
3- ุฏูุฉ ุงููุชุงุจุฉ ูุงูุตูุฑ ุนุงููุฉ ุฌุฏุงู ุฌุฏุงู ุฌุฏุงู
4- ูููุงูู ุจุนุถ ุงูู
ุนููู
ุงุช ุชู
ุชูุถูุญูุง ุจุดูู ุชูุตููู ุฌุฏุงู (ุชูุนุชุจุฑ ูุฏู ุงูุทุงูุจ ุฃู ุงูุทุงูุจุฉ ุจุฅููุง ู
ุนููู
ุงุช ู
ูุจูู
ุฉ ูู
ุน ุฐูู ุชู
ุชูุถูุญ ูุฐูู ุงูู
ุนููู
ุงุช ุงูู
ูุจูู
ุฉ ุจุดูู ุชูุตููู ุฌุฏุงู
5- ุงูู
ูุฒู
ุฉ ุชุดุฑุญ ููุณูุง ุจ ููุณูุง ุจุณ ุชููู ุชุนุงู ุงูุฑุงูู
6- ุชุญุชูู ุงูู
ูุฒู
ุฉ ูู ุงูู ุณูุงูุฏ ุนูู ุฎุงุฑุทุฉ ุชุชุถู
ู ุฌู
ูุน ุชูุฑูุนุงุช ู
ุนููู
ุงุช ุงูุฌูุงุฒ ุงูููููู ุงูู
ุฐููุฑุฉ ูู ูุฐูู ุงูู
ูุฒู
ุฉ
ูุงุฎูุฑุงู ูุฐูู ุงูู
ูุฒู
ุฉ ุญูุงูู ุนูููู
ูุฅุชู
ูู ู
ููู
ุฅู ุชุฏุนููู ุจุงูุฎูุฑ ูุงูุตุญุฉ ูุงูุนุงููุฉ ููุท
ูู ุงูุชูููู ุฒู
ูุงุฆู ูุฒู
ููุงุชู ุ ุฒู
ูููู
ู
ุญู
ุฏ ุงูุฐูุจู ๐๐
๐ฅ๐ฅ๐ฅ๐ฅ๐ฅ๐ฅ๐ฅ๐ฅ๐ฅ
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ุตุญู ุงููุฑุงุกุงุช ุงูุนุดุฑ ุฃุนุฏ ุฃุญุฑู ุงูุฎูุงู ุณู
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ุตุญู ุฃุญุฑู ุงูุฎูุงู ูููุฑุงุก ุงูุนุดุฑุฉุฃุนุฏ ุฃุญุฑู ุงูุฎูุงู ุจุงูุชูููู ูุตูุง ุณู
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immunfluoresence
- 2. SPECIFIC OBJECTIVE โข DEFINITION OF IMMUNOFLUORESCENCE โข TYPE OF IMMUNOFLUORESCENCE
- 3. DEFINITION OF IMMUNOFLUORESCENCE ๏ This test is similar to ELISA . ๏ A process in which flurochromes or fluorescent dyes are used which is marked with antibodies and exposed to ultraviolet ,violet or blue light to make them fluoresce.
- 4. CONT.. ๏ It use for detect antigen and antibody in their test use fluorescent dye known as โFLUORESCEIN ISOTHIOCYNATE. The method is suitable for only qualitative reaction this test is two type. ๏ Fluoresce is the property of absorbing light wave on particular wave length and limiting rays with different wave light.
- 5. CONTโฆ ๏ Procedure-:fluorescent dye coupled molecules without changing the antibody capacity bind to specific antigen. ๏ A antigen can also be labeled with fluorescent dye.
- 6. TYPE OF IMMUNOFLUORESCENCE ๏ 1.Direct immunofluorescence (unknown antigen) ๏ 2.Indirect immunofluorescence (known antigen)
- 7. DIRECT IMMUNOFLUORESCENCE ๏ In this test use specific antibody labeled ( single layer) with Fluorescent dyes. ๏ It is use for direct of unknown antigen in a specimen. ๏ It antigen is present react with labeled antibody fluorescence can be absorb use light.
- 8. CONTโฆ.. ๏ The fluorescent antibody technique is used for:- ๏ Rapid serological diagnosis of of bacteria ๏ Detection of ant toxoplasma antibody ๏ Detection of virus in cell.
- 9. CONT.. ๏ Procedure ๏ Prepare the sample by fixing it to the glass slide. ๏ specific fluorescein-labeled antibody to cover the fixed smear. Smear should not be allowed to dry during staining process. ๏ Incubate at 36ยฐC for 25 to 65 minutes in a humidified chamber ๏ The excess antibody from the smear by rinsing with buffer (e.g. phosphate-buffered saline) ๏ the use of a UV light microscope for yellow green fluorescence.
- 10. USE ๏ Detection of respiratory syncytial virus (RSV) antigens in nasopharyngeal aspirate ๏ Detection of rabies virus antigen in the brain of animals suspected of having rabies infection
- 11. RESULT ๏ Positive: Specific apple-green fluorescence ๏ Negative: No or little nonspecific background fluorescence
- 12. CONT..
- 13. CONT
- 14. CONT..
- 15. INDIRECT IMMUNOFLUORESCENCE ๏ In this test specific antibody labeled with fluorescent dye. ๏ It use for detection of antibody in serum. ๏ If during immunofluorescence test two antibodies are used then this known as indirect immunofluorescence test.
- 16. CONT..
- 17. CONT.. Procedure- ๏ A known antigen is fixed into slide. ๏ If specific antibody is present it react with antigen and make a antigen-antibody complex . ๏ Now fluorescent labeled antibody is added with react with fixed antibody. ๏ Incubate then wash and now observe under fluorescent microscopic. ๏ Fluorescent indicate presence of specific antigen and antibody.
- 18. CONT..
- 20. THANK YOU