i want to stain rat brain for ischemia. what would be the best procedure to avoid disturbance to the infarcted area? they day transcardial perfusion fixation denatures the tissue in place? would this be destructive in terms of interference with measurement of area of infarct?
Any procedure that uses formaldehyde will affect total volume of your sample, thus both post-fixation by immersion or perfusion will have this effect.
But generally speaking, perfusion will allow faster penetration of the fixative with better morphology conservation.
Anyway, the bigger impact in volume will be if your protocol includes ethanol dehydration.
Any procedure that uses formaldehyde will affect total volume of your sample, thus both post-fixation by immersion or perfusion will have this effect.
But generally speaking, perfusion will allow faster penetration of the fixative with better morphology conservation.
Anyway, the bigger impact in volume will be if your protocol includes ethanol dehydration.
The answer from David Rotllant is spot on. I would always choose perfusion if possible. Dehydration and fixation protocols vary from lab to lab, and this is why is difficult to compare absolute infarct volumes from TTC stainings, even between experiments done in the same lab. I suggest to express your results as a percentage of the total volume of the sample.
Seconding the answers of David and Federico I would like to point to RG-Archives by < SEARCH > function, pasting in your browser:
https://www.researchgate.net/search.Search.html?type=question&query=Immersion+perfusion+fixation or
https://www.researchgate.net/search.Search.html?type=question&query=Immersion+perfusion+fixation+infarction+area ,
respectively. It might be that you’ll find some answers to your query.
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