Author Topic: TEM and Immunohistochemistry  (Read 4675 times)

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Offline dr_meenakshi1501

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TEM and Immunohistochemistry
« on: August 19, 2003, 10:28:25 PM »
Hi,
I have learnt the basic immunocytochemistry and TEM.I am working on nerves and looking at the myelin.I wanted to know,if there are any antibodies which can actually detect myelin only??????
Will the ultrastrucure get spoiled if I use TEM after immunostaining?
Is it better just to use TEM without immunostaining?
Is there any difference in the fixatives for both????????

TEM and Immunohistochemistry
« on: August 19, 2003, 10:28:25 PM »

Offline richard03

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TEM and Immunohistochemistry
« Reply #1 on: August 19, 2003, 11:32:56 PM »
There are many ways to detect myelin. The most popular one is Luxol Fast Blue method at LM level. Check here http://ihcworld.com/_protocols/histology/luxol_fast_blue.htm

If you would like to detect myelin at EM level, the immunoEM is not necessary since myelin is clearly seen and easy to identify at EM level. Check these:
http://www.uni-mainz.de/FB/Medizin/Anatomie/workshop/EM/externes/Wartenberg/mhNerv3.html
http://www.uni-mainz.de/FB/Medizin/Anatomie/workshop/EM/externes/Wartenberg/mhNerv4.html
http://www.uni-mainz.de/FB/Medizin/Anatomie/workshop/EM/externes/Wartenberg/mhNerv7.html

If you do routine EM, use 4% paraformaldehyde and 1% glutaraldehyde in PB. If you do immunoEM or both, be caution about glutaraldehyde since it may inhibit some antibody staining.

Hope this helps.

Richard

TEM and Immunohistochemistry
« Reply #1 on: August 19, 2003, 11:32:56 PM »