I am beginning to study freshwater fish gonads and willing to use x-ray fluorescence and x-ray tomography. For the first one I am thinking in to use lyophization. But for the tomography I am considering to fix the samples with formol 10%. Do anyone know if there is a better preservative substance for the gonads and the technique, and if anyone could help me with sources about this matter I would appreciate.
I assume the samples are fresh, i.e. no staining etc. applied?
Did you consider using algoras to embed the samples?
Alexander Rack thank you for the reply. Yes, they are fresh. We are fixing it in formol, (metanal), and intend to wash it with water or ethanol. The problem is that the fish migrate and we were not prepared to receive the samples, but they do not wait! I am considering in to use the Karnovsky solution, but I need to check if it will be possible to be made in the lab.
I do not know about algoras, if you could suggest some references I would very much appreciate.
Sorry, typo - it is agarose , I never used it myself but there is a lot in the web
https://www.youtube.com/watch?v=KKmiKKMDDhY
I am sure I have also seen soft tissue scanned without any fixation. For storage then indeed more care has to be taken, i.e. cooled storage etc. is required.
Marie-Christine Zdora and Alexander Rack We've decided to fix the samples in formalin and are considering embedded it in agarose. Thank you very much!
We have scanned cochleae and many other types of tissues before and after decalcification using synchrotron radiation. Tissue can be fixed or fresh (fresh requires shock freezing) and thawing at the beamline. This affects tissue quality. Conservation can be done with any fixation method. Depending on what you intend to do after the x-ray tomography you have to decide on the type of method. Agar embedding may cause problems. Agar acts as a phase scrambler. With thick layers of agar around the samples, the image quality suffers. Scanning in fluids however is possible. For the latter approach, you want to degas the fluids.
Edison Freitas Cossignani, I suggest you read the paper:
The paper reports measurements carried out in a synchrotron facility. The samples were fixed in formalin 5%. For tomographic imaging, they were washed in distilled water for 10 minutes, and subsequently, each sample was dehydrated in a graded ethanol series and the samples were transferred into polypropylene pipette tips containing pure ethanol (we had success in the use of isopropanol, ass well).
A key point was a movable plunger was used over the pipette to increase, though little the pressure under the ethanol avoiding the appearance of bubbles.
If you are using a desktop system I suggest you try stain with iodine solution .
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