Hi all,
I've been working on a project that involves microinjecting cells into zebrafish.
A big issue has been needle clogging. Once capillaries are loaded, cells clog the tip of the needle and it's nearly impossible to inject a standardized/consistent number of cells in each fish.
ANY tips would be much appreciated. I'm wondering if a shorter or wider needle would work any better? Right now I am using 6 inch non filamented thin-walled glass capillaries 1.0 OD/0.75 ID.
The capillary absolutely cannot have the filament - that shreds the cells. Larger needle opening will help as well. Also, you can add a little of DNaseI to the cell suspension - that will prevent cell clumping due to the released genomic DNA.
Aidas Nasevicius
Wonderful - thank you so much.
If possible, would you be able to share the type of capillary you use?
I use the same needles for my injections. I have had this problem as well before. Do you chill your needles before injection? Putting your needles in the fridge or on ice prevents cell clumping and is the easiest solution if you have not already tried this. I have had to cut my needle back for a larger opening--it means more damage to the fish so I avoid it. It may also depend on the cell line you are using. Is it prone to clumping? If so, you may need to add something to your suspension to prevent clumping. You could also turn up the pressure to clear the tip with force.
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