I want to do immunohistochemistry on an aorta. I want to store in 70 °C, but the 70 °C refrigerator is full, so I think I need to store in a liquid, but I don't know what liquid to use. My friend told me to use PBS, another told me to use the buffer formalin. Thanks for your help.
I would suggest you to formalin fix the tissue first for overnight and then store in 70% EtOH at 4 degC for long time.
There is another active parallel thread is available addressing similar question at
https://www.researchgate.net/post/Storage_of_tissue_for_immunohistochemistry
Hope that helps.
Consider first: How do you want to cut it afterwards? Which antibodies do you want to use? Fixation like formalin or ethanol might change your epitopes. Working with formalin-fixed tissue is the standard and paraffin-embedded blocks can be stored at 4°C or room temperature, but antigens might be destroyed during fixation, since formalin modifies the amino acids. For cryosectioning it's best to shock-freeze the organs right away and keep them cold (-70°C or liquid nitrogen). So look in your antibody data sheet, if your antibodies are made for use in formalin/PFA tissue or only with cryosections.
Don't store you organ for much more than a day (better only a few hours) in PBS, it will loose morphology and antigen reactivity.
Decide first what you are going to do with the tissues later. Fixation in formalin, Paraformaldehyde or gluteraldehyde helps immunolabelling with some antibodies and on the other hand it could mask the antigenecity.
If it is a short term storage 'RNA later' would be good (you can find a recipe online to prepare in your lab or, can buy it commercially).
For the full tissue, you can store in sucrose 30% after fixation. If you want to store slices (e.g. microtome-sliced), you can use 'Antifreeze buffer':
Antifreeze
2 L :
57.5 mL Buffer A
192.5 mL Buffer B
750 mL H2O depc
600 mL Ethylene Glycol RNase free
400 mL Glycerol RNase free
Buffer A
1 L :
27.6 g Sodium Phosphate Monobasic
Complete with water
Ajust to pH 7.3,
Autoclave
Buffer B (antifreeze)
1 L :
28.4 g Sodium Phosphate Dibasic
Complete with water
Ajust to pH 7.3,
Autoclave
For IHC You can keep the tissue in formalin for few weeks with no problem. But if your talking months or years, your best choice would be to generate paraffin-embedded tissue which you can store in a relatively controlled environment (I use a regular refrigerator for this) for so many years. I've used blocks that are 65 years old with no problem. Any pathology lab would do this for you just make sure to let them know you only need paraffin-embedded tissue for the time being and NO sections.
You cannot "store an organ" (which organ ? each one has its features / must first learn some elementary data before immuno-...) in a liquid. In morphology, we use different names for different receipts of mixtures, according to the purpose - including immuno- ... And another qu - immuno- for what ? And how long do you want to store it ?
I want to store about 2 months, the organ is rat aorta,,,by the way thanks for the answers,,,,
- If yor organ is already fixed by perfusion, you can let it in the same fixative for 3 days. then, if you want to embed your tissue in paraffin, it's better to transfer it in PBS at 4°C and keep it for many months. You have to change PBS solution each two weeks or use sodium azide. If you intend to cut the organ by cryocut, you have to transfer it into sucrose 20-30%. you can store it in this solution or transfert it to PBS (but after a minimum of 2-4 days in sucrose).
What book i have to read about this? And what basic science learning about it?
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