I dissected and stored the tissue at -40 and it's been around 5 days, can it be used for TEM imaging. If it can, before using fixative what precautionary measure i must adhere to?
If you are interested in something like collagen, then you may have some luck (collagen is almost non destructive). However cells could be badly damaged by ice crystals. You may try. You' may see something, may be even something useful, but do not expect publication quality pictures.
Thank you Vladimir, I was more inclined towards mitochondrial dysfunction. I reckon it won't provide any significant insight or image.
I just realized I have sample in dissolved formalin for histopathology, can those be washed with buffer and used for TEM ?
i think that fresh samples are prefered and more clear because formaline can effect on the tissue
Formalin-fixed tissue samples can be used for TEM if well washed and refixed with glutaraldehyde and OsO4. The quality of TEM images will highly depend on the elapsed time between the interruption of oxygen supply to the sampled organ and the beginning of the fixation procedure (duration of anoxia). The size of the tissue blocks (as they are usually large for histopathology) may determine the quality of ultrastructure as well. Take the most superficial zones of histopathology blocks for further processing.
Frozen samples may have tissue artefacts caused by ice crystals at cellular level, that look like pale empty spaces inside the cells cytoplasm cutting randomly membranes of cells and organelles, but not all mitochondria may be targeted by these mechanical artifacts. The time factor (as mentionad above) is again the most critical factor for preservation of mitochondrial ultrastructure. Good luck.
thank you , Barakat Abd El Maleck and Dimitrolos Krajci .
Fresh samples is not possible, but i will certainly try TEM of formalin fixed sample after following what suggested by Dimitrolos . Let's see how it goes.
Regards
- Badges
- Science topic
- Similar topics
- Biological Science
- Anatomy
- Tissue