Hi,
I have a hydrogel scaffold seeded with schwann cells to study myelination process.
now I want to fix it and see it with transmission electron microscopy.
i've read some articles mainly about in vivo fixations; but there are few articles about this special case and i need help.
Thank you in advance
Can you can get cell suspension (removing cells from a scaffold), then follow protocol suggested by Anand Patwardhan. If you cannot remove cells, or want to observe them while on a scaffold, you will need to embed your specimen in a resin (follow standard TEM protocol for tissue).
By the way, both cell suspensions and bulk tissue specimens can be dehydrated with alcohol or acetone; alcohol is used more often.
When speaking about in vivo/in vitro fixation, we speak about bigger organisms, like mouse or elephant. For individual cells it's always in vivo fixation.
Basically you are looking for protocol to fix Cells my EM. Yes there is different between Cells & tissue fixation protocols for EM. Cells are dehydrated by using ethanol while tissues by acetone. Do you want detailed protocol ?
Good luck.
Hi Vladimir
it would be vary nice if you tell me the differences like dos and don'ts.
Thank you
Its very similar to what we do...http://www.ihcworld.com/_protocols/em/em_routine_cell.htm
it does use hazardous chemicals. So take precautions !!
Can you can get cell suspension (removing cells from a scaffold), then follow protocol suggested by Anand Patwardhan. If you cannot remove cells, or want to observe them while on a scaffold, you will need to embed your specimen in a resin (follow standard TEM protocol for tissue).
By the way, both cell suspensions and bulk tissue specimens can be dehydrated with alcohol or acetone; alcohol is used more often.
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