I'm assisting in a light microscopy study where I count the number of pollen grains in a sample, separating normal grains from malformed grains. The pollen are suspended and stained in cotton blue. The goal is to see if notable normal:abnormal pollen ratios exist among hybrid plants.
I was instructed to vortex the solution for uniform mixtures. However I am concerned that vortexing may make shear the normal grains' outer casing, release their contents, and erroneously make these grains seem abnormal.
Dear Alwynne,
Thank you for the information! The pollen I'm using were collected from living plants, and were stored for at least 6 mos in cotton blue at room temperature. No other preservatives were used. And yes, by "malformed", I was concerned that vortexing would rupture or tear their exine, and alter the pollen's internal structures. In short, I'm comparing the quantity of viable pollen against non-viable pollen.
Cheers,
Gary
Please have a look at this link for the answer.
https://www.britannica.com/science/pollen
One of the gold-standard procedures for identifying pollen grains, acetolysis (Erdtman, 1952), involves high-speed centrifugation of pollen with no ill effect. I have used the method with a vortex, and did not detect any negative effect on pollen integrity.
I did, however, rupture some of the very large pollen grains (esp. Iris spp.) by placing the cover slip on too hard
http://www.tandfonline.com/doi/abs/10.1080/11035895209453507
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