Many thanks for the positive feedback. Since we’re a teaching laboratory we don’t often get access to fresh lymph nodes but this is a great idea for future videos.
Thanks Na. Glad you found the video useful. If you are interested, additional content on frozen sections can be accessed from my online course by joining my channel as a “trainee” member.
How is the frozen section held across the blade? like in paraffin sectioning you have the wax covering the entire specimen, giving it a rectangle shape, since no such embedding is done in cryotomy, do you just place the whole thing in the holder?
Spheroids can be very tricky, especially when small. My suggestion would be to suspend multiple spheroids in agar following fixation and before processing into OCT. You will also need to be careful when sectioning but embedding multiple spheroids per block should ensure that you achieve cross sections through the middle.
Have a great day sir. I have a question: with spleen, lymph node specimens, should the section after being embedded in paraffin be cut thinner or thicker than usual ? And why we shoud do that ? Thanks for answering me !
I think this depends on a number of things. Lymphoid tissue is generally more fragile owing to less ECM components and higher density of cells, so you may gain advantage by cutting slightly thicker sections, but generally this issue can be addressed via use of picric acid in fixative solutions such as Bouin’s.
Hi James, most laboratories use a commercial embedding medium known as O.C.T. which is an abbreviation for Optimal Cutting Temperature. OCT contains polyethylene glycol and polyvinyl alcohol dissolved in water. You can buy OCT from a variety of scientific suppliers.
As an anatomic pathologist. Much appreciation for this video! Hope you can sample other tissue such as lymph node.
Many thanks for the positive feedback. Since we’re a teaching laboratory we don’t often get access to fresh lymph nodes but this is a great idea for future videos.
Your videos are very useful, I greatly appreciate them!
You are most welcome. Thanks for watching!
Thanks Damien and Chris, wonderful job!! very useful video for early learners!
Glad it was helpful!
Thank you so much! So well and clearly explained! Really, thank you!
Thanks Na. Glad you found the video useful.
If you are interested, additional content on frozen sections can be accessed from my online course by joining my channel as a “trainee” member.
Nice vid always wondered what that anti roll plate was for as ones at work dont have the glass piece
How is the frozen section held across the blade? like in paraffin sectioning you have the wax covering the entire specimen, giving it a rectangle shape, since no such embedding is done in cryotomy, do you just place the whole thing in the holder?
The section is held together owing to the frozen water present within the tissue. The ice is the equivalent of paraffin used in regular microtomy.
Hi. Thanks for this video. I am working with spheroids from cell culture . What is your suggestion for thickness? Thanks
Spheroids can be very tricky, especially when small. My suggestion would be to suspend multiple spheroids in agar following fixation and before processing into OCT. You will also need to be careful when sectioning but embedding multiple spheroids per block should ensure that you achieve cross sections through the middle.
5-8 microns would be a good starting point for thickness.
Have a great day sir. I have a question: with spleen, lymph node specimens, should the section after being embedded in paraffin be cut thinner or thicker than usual ? And why we shoud do that ? Thanks for answering me !
I think this depends on a number of things.
Lymphoid tissue is generally more fragile owing to less ECM components and higher density of cells, so you may gain advantage by cutting slightly thicker sections, but generally this issue can be addressed via use of picric acid in fixative solutions such as Bouin’s.
@@damienharkin yeah i got it. Thank u so much.
ASCP 30 years fromTexas,very nice technique
Thank you Sir.... It's really helpful
So detailed, Thank you
You’re welcome 😊
Hello, I wonder will you dry out your slides at Room temperature after cutting?
Very helpful. Thank you very much
Amazing 🤩. Thank you so much.
So educative
What is the embedding medium for frozen sections called?
Hi James, most laboratories use a commercial embedding medium known as O.C.T. which is an abbreviation for Optimal Cutting Temperature. OCT contains polyethylene glycol and polyvinyl alcohol dissolved in water. You can buy OCT from a variety of scientific suppliers.
Negative -50
very well, thank you very much
does tissue sectioning for ovary also uses cryostat?
What is the thickness of sections ?
The thickness varies a little according to the tissue but generally we cut around 6-8 microns.
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