Dear All,
I have already seen many Q&As regarding HistoGel handling and tried various changes of the dehydration protocol. Sadly, nothing has worked out so far.
I am currently working with spheroids and use HistoGel for Histology, since it would be very difficult to handle the spheroids otherwise. During dehydration, the gel becomes really hard and makes paraffination almost impossible. While embedding in paraffin, the gel remains like a single unit within the paraffin block and does not really unify. While slicing later, this unit falls sometimes off after transfering on the water, or the slicing does not work at all (paraffin brakes completely). Does anyone know how to avoid HistoGel hardening? I would appreciate any tricks you have up your sleeve. Maybe also not involving HistoGel ;)
Thank you very much!!
Best regards,
Astrid
Dear Matthew Ibbs
Thank you very much for your answer! Since I have just started doing Histology, we do not really own a tissue processor and I am doing everything "by hand". The dehydration protocol does not include acetone but xylene. I start with 50% EtOH and go up to 80%, then 96% and finally 100%, before going to xylene. Each step 15 min. After that, I go into paraffin:xylene, then paraffin for an hour and finally paraffin over night. When I open the cassette for embedding, I find a very hard HistoGel.
As you said, sadly in the instructions of HistoGel there are not really any helpful informations.
Best regards,
Astrid
Dear Matthew Ibbs
oh! I really have not thought about that. I have changed a lot in the alcohol times and concentrations, but not the paraffin itself. I will definitely try this out the next time!
Will let you know.
Thank you so much for your insight!
I am trying to go through this process as well. I have tried your protocol Matthew and it gets hard in the xylene process.
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