I have 33 uL samples, and need to transfer 10 uL of each sample into a 384 well plate. After doing this, I often find air bubbles in my samples in the 384 well plate, and this interferes with later aspects of the protocol. I am using 20 uL tips, making sure there are no air bubbles in the tips, pipetting slow, and holding the pipette in place after expelling the necessary volume. I am also taking care to stop pipetting at the first stop, and not the second stop (using the reverse pipetting technique). However, I am still struggling with frequent air bubbles in my samples. Is there any alternative pipetting technique I might try, or something else I should be cautious of during this process? Or is there something I might try to get rid of the bubbles after they form?
No experience for Cell free techniques .. but i know a laborious method
if your samples not heat sensitive you can use flame out after pouring samples to well just use direct flame over well .. i have used this technique in agar plating ..
or if they are sensitive you can use sterile inoculation loop or sterile needle and heat it up and manually just move that needle over the sample to remove air bubbles .
you can go through :
http://blogs.rsc.org/chipsandtips/2017/02/28/a-simple-bubble-free-cell-loading-technique-for-culturing-mammalian-cells-on-lab-on-a-chip-devices/
All the Best
Did you try anti-foaming agent ? probably it is Ok since it is neutral
https://handling-solutions.eppendorf.com/.../pipetting.../how-to-pipett...
Amir Pandi I will definitely give that a try, thank you very much for sharing!
To get rid of bubbles after they form, try popping them with ethanol vapor -- just take the straw out of a squirt bottle, fill with ethanol, and blow gently on the bubbles.
Hi,
THe approach suggested by Augustus could be followed. Prof. Khaled's advise to use antifoam could also be considered.
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