Dear colleagues,
How to mount nematode for SEM? I would like to mount the nematodes without they are laying down in the stickly black surface. If the nematodes is sticked it, I dont get to see their structures :( !! Thanks :)
Chapter Techniques for preparing nematodes for scanning electron microscopy
@Vladimir Dusevich I red it. Thank you!! Do you known other strategies for preparing nematode for SEM? For example, somebody use hair. What is your experience?
I am sorry, I have no experience of working with nematodes at all. As for hair... Do you mean an "eyelash", a tool that looks like a painting brush with just one hair? Of course it is useful for manipulating a tiny objects when mounting SEM specimens.
Well, you can adhere nematodes on poly-lysine coated Thermonax plastic coverslip after sputter coating or at dehydration steps.
Hi Carlos,
We normally use the following protocol for taking LT-SEM images of plant-parasitic nematodes:
We place individual nematodes on a smooth Millipore membrane filter containing 0.4 µm pores, and we remove the water excess through the membrane filter from below with filter paper. You can place a variety of elevated structures, as well as the standard filter paper on the membrane in order to get a good face view of the nematodes. Next, the membrane filter containing the nematodes is affixed to a copper plate, using conductive carbon tape and a small amount of cryo glue, and conductively frozen for 30 s on a pre-cooled (-196 °C) brass bar partially submerged in liquid nitrogen before fully submerging the samples into liquid nitrogen.
I hope this helps!
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