filling micropipettes free of air bubbles

Dirk Hampf d_hampf at web.de
Tue Aug 20 10:07:52 EST 2002


Hi!
We want to isolate cytoplasm of single cells (lymphocytes, lymphoblasts) in
order to measure the concentration of some drug metabolites. For this
purpose we use an experimental setup similar to patch clamp or ICSI. With a
micropipette single cells are tapped to "suck" out the inside. We have the
following problem: The micropipettes (inner diameter 1-5µm, borosilicate
glass) should be charged free of blow holes with mineral oil. Attempts with
purchased micropipettes with an inlying filament as a filling aid have
failed so far. Currently we are pulling the pipettes independently (Sutter
Instrument P-97 micropipette puller). We have not been able to fill the
micropipettes free of air bubbles. We already tried to fill the capillaries
with oil by using  over and low pressure without any result. Please send us
any ideas, we would be very thankful.

Regards
Sandra Kalich, Dirk Hampf





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