I am planning to do immunofluorescence (IF) and immunohistochemistry (IHC) from rodent tissues. Is it fine to perform IF and IHC from tissue sections from another tissue of same animal group that was not used for H and E Staining ? I have seen in many papers that they use the same tissue section (in continuation while sectioning the block) for H and E, that was previously used for H and E Staining.
If used another, will the reviewer ask the question on this issue ?
Hi Jagtar Singh
Yes, you can use any tissue from the same group to perform IHC, IF or H&E. It is not necessary that all the staining has to be performed from one animal tissue of the group.
Hope it helps,
Thanks,
If you are preparing serial sections of your tissue block (paraffin as well as free floating frozen sections) you can use alternating sections for different stainings and immunohistochemical reactions with different antibodies.
If you like to mount your sections directly on the slides then you have to prepare a schedule of stainings . That means lable the slides for different stainigns and then mount the sections. Example: Slide 1 HE, Slide 2 IHC antibody A, Slide 3 IHC antibody B. Good luck!