Does normal saline (0.9% NaCl in H2O) have a limit of support for fresh biopsies meant for immunofluorescent stain in terms of length of time?
Saline does not 'support' living tissue; it is a salt solution, without any of the essential elements that support life!
The purpose of saline is to prevent dehydration and isotonic shock.
Different tissues will have different tolerance for saline before exhibiting the various manifestations of cell death. Chilling samples (
Hi Kamoru,
I agree with David saline (0.9% NaCl in H2O) can damage fresh biopsies.
You can fix tissue first in 10% Neutral-buffered Formalin for some time like 24hours (depends on thickness of tissue), if too thick, can prolong time. Take tissue out and wash 5 minutes 3 times in PBS and place in PBS and keep at 4%. for few days. Or best is start dehydrating tissue in 50% ethanol and store in 70% ethanol for many days at 4C
David Ian Leavesley Prof., I really appreciate your contribution by providing scientific reasons furthered with a better choice in replacement. Indeed, we have been using NS with refrigeration as you added and it has always been helpful without affecting the outcome. It is just that scientists should not wait to think ahead in case of any harsh situation to offer a solution. Your reminder greatly helped. Thump-up!
Walid Hassene Hamri No, it cannot damage it unless all metabolic requirements are exhausted. Be reminded that normal saline is also known as physiological saline, it is isotonic and not the same as putting salt crystals in food. Normal saline has the same osmolarity as plasma, and if it can damage fresh tissue it means you and I cannot be living by now as humans.
Normal saline can be directly administered to the bloodstream without damaging our blood cells or tissues.
Subhash C. Juneja thank you for providing more insight!. Be reminded that formalin causes non-specific fluorescent signal, are you sure this will not aid false positivity?
Your initial question was storage of tissue, you did not metion which way you want to do your immuno. If you want immunofluorescence, you better do cryopreservation and cut cryo sections, if you want immunohistochemistry using non-fluorescent method, than you go for paraffin sections, I will send you 2 reprints tomorrow for these procedures,
Dr. Subhash C. Juneja thanks for this contribution, thumb-up! Meanwhile, the question, in bold, mentioned: "...meant for immunofluorescent stain". I would appreciate other methods, it's gaining more knowledge from scientists like you, thanks.
Yes, you can do cryo sectioning of tissues, means you have to freeze tissue in Liquid nitrogen after fixation, and also you do not need antigen retrieval in on cryosections, rest immuno procedure is similar to other system like paraffin sections, good luck
https://www.urmc.rochester.edu/musculoskeletal-research/core-services/histology/protocols.aspx
Please get good knowledge of protocols from this site, if you have any specific questions, you can ask me
Emmanuel Sunday Okeke , Stanley Ebhohimhen Abhadiomhen and Charles Obinwanne Okoye thank you for your comments.
Check https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4016408/
Chinaza Godswill Awuchi thank you for your contributions, the two articles are highly informative, thumb-up!
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