I want to dissect damselfly specimen and wants to count the number of ovarioles and eggs to estimate the fecundity status of damselfly. I am not an entomologist and have almost no experiences of specimen dissection. I have go through some articles but could understand the protocol. I am expecting an elaborate protocol which will guide to do the job. Thanks in advance
sir, this paper may help you.
https://www.researchgate.net/publication/50886588_Number_and_Size_of_Eggs_in_the_Three_Emerald_Damselflies_Lestes_sponsa_L_temporalis_and_L_japonicus_Odonata_Lestidae
I've not worked with Damselflies, but I have worked with other insects on egg counts. This will depend on how easy it is to break the ovarioles away from each other and as such would only likely work in the latter stages of oogenesis. You can place the ovaries into a set volume of saline - call it 100µL - and try vortexing or pipetting up and down to break the ovarioles away from each other. Too much vigorous mixing might destroy eggs though so be wary. You may have to tease them apart with forceps or dissecting needles if they're particularly fragile. If you take say 10µL of this fluid onto a microscope slide and count all of the eggs you can infer that this represents 10% of the eggs in the specimen. You should make sure the eggs are mixed into the water thoroughly to ensure you are not taking just water or just eggs and you should count at least three times to get an average.
I've done this with reasonable effect in mosquitoes, but I confess I can't be sure how well it would work in damselflies. What are you counting the eggs for? In the past I've used the bradford assay to measure the amount of protein in a sonicated ovary to score fecundity which is also a good measure that doesn't involve counting so much
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