Monday, 14 July 2014

Battle 2 with the Electron Microscope.....

My second battle with the electron microscope took place last Thursday, and I can happily say that it was a successful one!!

I tested a few different staining techniques as my last images would not focus because of the techniques used for staining.

My first image was staining with uracil acetate 3 times but blotting straight away. These were the results...

Much better than the previous trail, as you can now see some of the more detailed features of the virus and see that it is in fact hexagonal. However the background is pretty salty! But better was still to come!

My second staining method was using uracil acetate but leaving the sample to stain for 30 seconds, unfortunately this method didn't give any images that were in focus so it was concluded that this staining method was not successful.

The third method I tried was leaving the stain for 1 minute. Here are the images...

 I think these are the best images of TCV that I took, and the most successful staining technique!

When I left the stain for 2 mins, the sample was slightly over stained, but still in focus!

Some of the detail can still be seen, but the virus capsids appear a little bright in contrast to the background, indicative of over staining. 

So I have found out that the best staining method, for me anyway (there are loads of different protocols that say otherwise with different methods), is to leave the uracil acetate on the grid for 1 min before blotting off the excess. 

All in all a pretty successful ending to a unsuccessful week!  

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