I have always worked with whole tissues. Now I will work with cells. Granulosa cells will be obtained form follicular fluid (from follicle aspiration) after centrifugation. As I have never worked with these samples, I´m looking for a protocol to collect them in a slide, fix them, preserve them for a month and then perform an immunocytochemistry. thanks!
You can make a cell block the technique you can found everywhere.
But in my experience, instead of using commercial gel (ex Histogel), we use Agar gel 3%, it works well.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3148935/
If you have access to a cytospin or thin prep technique, you can use these for concentrating cells in fluid specimens and can fix with whatever fixative you need to use for the staining that is to be applied.
Hi Florencia,
The approach is dependent of what kind of cell population you are interested in. If your primary goal are the sao called "rear events", preparation of cytospins with a cell concentrator is a good option. If you want to preserve the material for like 5-10 years, a preparation of a cell block from a cell pellet is also a variant. The pellet has to be afterwards fixed which might result in certain loss of analytes.
As far as I got you need to have a spin preparation. The spins can be stored dried in a fridge for about a month w/o fixation. Add dessicant (silicagel for example) to the box and seal it well to prevent yeast grow on the cells. The slides can be stored in a fridge for 1 week to 3 months according to my personal experience. For longer storage the spuns can be frozen but this kills the morphology completely.
Spin preparations can be performed in any cell spin centrifuge (I hope you have one in hand). The collected fluid can be directly spun down to a slide or can be diluted in PBS to a certain volume. I will suggest that you wash the cells couple of times before making the cell spin.
These are very general suggestions. I can dig out a whole protocol if you're still interested.
Best,
Ivan
Either you can do cellblock from the fluid & do immunohistochemistry
Or do immuno stain on the fluid it self
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