Cryo TEM sample preparation using Vitrobot

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Sit back and enjoy the show!
If you want to give credit, please give credit to the Cryo EM facility of the National University of Singapore.

[@borjaabascalimizcoz8248]

Thanks very much for the video!! Its very helpful!

[@tongsheng00]

It helps a lot. Thanks.

[@BhanuMantri]

how you can set the lot time and blot force ? what are the parameters we should keep in mind to set these values?

[@MrRight2006]

Parameters depend on the nature of sample. You need to play with different parameters at first. Try with different blot time at first. Vary between 1 and 10 sec . Then try changing blot force. Sample amount is about 3 to 3.5ul. The trick in the transfer is that you need to have a stable hand during transfer. Rest your palm on the cup, release tweezers and then transfer.

[@MrRight2006]

you welcome!

[@buzy2a]

hi, could you give me the diffrent paramaters you use: Blot time, bolt total, blot force and the quantity of liquid you put on the grid ? And do you use a special trick to transfer the grid from the ethane to LN2 ? Thx a lot

[@MrRight2006]

welcome :)

[@user-oo4hv9ye7w]

3:28 4:43 6:22 6:32 6:51 7:47

[@frankthespank]

Will this help me sneak dinosaur embryos out of the park?

[@jingchuansun6915]

You didnot put anti-contaminator (a float ring on LN2). Why?

[@SofyaPolyanskaya1]

To let people see the details of the process obviously.

[@solarbuff350]

Great video. One question: When you take the sample out of ethane and into the liquid nitrogen, can you dip the tweezer in the liquid N2 as well? If you dip the tip of the tweezer too much, there will be bubbling because the upper portion of the tweezer is still at room temperature and I wonder if the ice will be formed on the sample because of this. I cannot see where your liquid nitrogen level is, so cannot be sure what you did at this step. Thanks!

[@borjaabascalimizcoz8248]

To avoid that you can dip the tweezer before taking the grid, or at least touch the edge of the metal cylinder. I hope this helps :)

[@PedroRodriguez-Amor-y-Paz]

Vitrobot FEI

[@teleportvalorant7658]

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