Dear Scientists
I need to capture the larvae for biochemical assays. whether I can use tricane or If I use Tricane to anesthetize also when pippeting the medium into 1.5 ml centrifuge tube for PBS wash, the larvae get enters into the tips. I am spending more time in washing and feeling difficulty. In what way I can improve my work. Can you suggest the alternative.
Using tricane on the embryos shouldn't mess with any biochemical assays, we use it to knock out our larvae for manipulations or collections. Just don't leave them in there for too long (~30 min) or they will die.
Have you tried using PBS-Tween instead of normal PBS? The Tween detergent helps to prevent the larvae from sticking to the inside of the pipettes.
I also recommend the Tween20 to help lubricate your pipette.
Do they need to stay in their own individual centrifuge tubes? If not, then consider using a Multiscreen TM 96-well microtiter plate (see Fig. 2 in the publication below). The bottom of each well is made of a fine mesh screen and you lower the plate into its buffer tray that contains whatever agent you wish to use. Because of the buffer tray and screen design, all wells will go through the same conditions. If that is OK then it really speeds up the washing process without the need for pipetting.
Hope that helps.
- Melissa
Article Adapting the Medaka embryo assay to a high-throughput approa...
Thanks Clay and Melissa
I didn't tried with Tween20. I will proceed and let you Know.
-Maharajan
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