Tissue Transport Mrdium

[fery]

Hi All


would you please let me know what is the best tissue transport medium for flow cytometry or IF staining and what is it's formula


Thanks in Advance,

Fery

[ImmunoNYC]

Transport from where to where? A far or short distance? How much time needs to be accounted for? I assume these are not frozen?

I would say if the tissue is UNFIXED and still alive I would transport it ON WET ICE in whatever culture medium is appropriate to the type of tissue with the aim of keeping it alive for as long as possible. As a routine action (if the information about specific media is not known) I use RPMI + 10% fetal bovine serum (plus antibiotics) or something similar to that. (RPMI and the serum are all commercially available - no need to make up yourself). The key is to give it goodies to keep it alive and keep it cold.

If the tissue is FIXED but not frozen you can transport in fixative or perhaps 30% EtOH if going a short distance. If it is going by plane you cannot send ethanol so I would send it in PBS or HBSS. Again to be cautious I would recommend sending on ice only to avoid any potential bacterial/fungal growth. (Both of these buffers are commercially available or can easily be made up - just google them for a recipe or look it up in any common lab reference book).

[fery]

Dear Maximina

Thank you very much for your reply. I need a Medium To keep unfixed  Lymphocyte cells in tissue sample alive for Maximum time possible and suitable for flow cytometry.



Regards,


Fery

[ImmunoNYC]

Are they already disaggregated from a primary tissue? Or is it a cell line?? Or is it a prep of tissue such as bone marrow, spleen or lymph node. This makes a difference as to my answer.

Broadly speaking in any case I think I would go RPMI or perhaps X-Vivo. Both are commercially available and are generally speaking suitable for culturing lymphocytes. You probably want to add some serum in there too.

You should know no tissue will stay alive for a very long time on ice in this manner without proper gas exchange and temperature.

Can you possibly freeze them in appropriate freezing medium and send them on dry ice?

[fery]

Dear Maximina


Thanks alot for your help


Best,

Fery

[excalibur]

Fery, I suggest using Michelle's fixative (actually a fluid and not a fixative)to keep the cells alive. Also, M-K (McCarey-Kaufman) Media, Optisol, and Likorol are used to keep donor corneal tissue alive during transport. Tissues the size of human corneas survive and have been transplanted with success as long as 7 days in M-K Media.

Most companies that sell fixatives, such as formalin, also have Michelle's.

Polysciences also has their own transport medium:
http://www.polysciences.com/shop/product.asp?dept%5Fid=203555&pf%5Fid=24311

[fery]

Dear Excalibur

Thank you very much for your Help. I appreciate you if you give me michels medium formula.


Best,


Fery