Hi all,
I am a graduate student at ISU working on the nervous/neuroendocrine system of various insects. While still succeful on my slide mounts, I am struggling with the best way to remove fat bodies clinging to the ganglion of my specimens - specifically Tenebrio molitor. Currently, my process is as follows:
Fresh vivisection, rinse specimen in Formalin, removal of digestive tract etc, rinse again with either Formalin or PBST + 1 or 2% Triton. Some manual removal of fat clinging to CNS - upon complete removal of the nervous system + brain from my specimens, I wash again with PBST +1-2% Triton and attempt to manually peel off remaining fat bodies and tracheal tubes from (typically) the thoracic ganglia and brain using the finest forceps I have in the lab. My little buddy T. molitor doesnt have a lot of tensile strength in the nerve cord, and it is therefore fragile and easily wrecked.
As one can imagine, this gets to be incredibly frusterating at times plus I am unable to completely clear my specimens from fat - which sometimes interferes with fluorescent microscopy post-processing. I also cannot allow my specimens to dry out because I am tagging neuropeptides with fluorescent antibodies.
Maybe I'm screaming into the void here, but does anyone have any good techniques or tips they're willing to share? I just want to get the best imagery I possibly can and minimize non-specific binding.
Thanks!
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