I am extracting hemolymph from bees and using Giemsa stain so that I can differentiate and count hemocytes. Do I need to apply a mounting medium and coverslip to my slides after staining to preserve my samples? If so, what type of mounting medium should I use?
I see that Qayum et al., (2011) uses PolyMount or Shur/Mount, so I'm guessing that a
toluene-based acrylic resin would be ideal? I am still not sure if I can access these materials, so it would be great to know if there are viable alternatives. Thank you!
Dear Maggie,
IMO you should mount the cells for better results.
It is possible to do microscopy without mounting a coverslip, when you use oil immersion, but I did that never before and read it only in older papers. And I guess it´s a mess.
For proper mounting: After Giemsa staining air dry the slides, then place them in Xylole (or suitable xylole replacement) for 30 min to overnight, let the xylole vape off, and mount with DePeEx (DPX) or Entellan or similar mounting medium.
You may try to mount them with water or glycerole based mounting medium, but again I have no experience with that. I´m not sure if Giemsa is stable in water or glycerole or if the stain will wash out. That´s something you have to test before you loose precious specimens.
Be carefull, because Giemsa staining for Cold-blooded vertebrates should be blot ,dry and mounted , but Giemsa for birds says clearly do not blot.
See Humason GL, Animal tissue techniques Freeman co 1967. Bees ????
Is there a histolab in the vicinity? They have this reagens for routine work. I don't recommend water-based mounting media for Giemsa. The stain would bleed out.
A few dips in 100% ethanol and then a few minutes in xylene or substitutes would do as preparation for mounting.
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