Multichannel Systems HiClamp Manual User Manual
Page 80
HiClamp Manual
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Fine selection
Use a stereo microscope and the golf-club shaped glass tool to check each single oocyte
for the following criteria.
Outer form:
No visible damage of the cell.
Well separated colors (dark and light brown).
No residues of follicular tissue.
Size:
Uniform size of about 1.2 mm.
Note: Selecting oocytes is an important step. Perform it very carefully to obtain best results.
11.3.6 Plating Oocytes
You need a well plate filled with Barth's + gentamicin.
Important: Visually check all well plates before use. Use only unsterile and very flat plates.
Sterilization may lead to warped plates. Warped plates will decrease the injection and expression
efficiency.
1. Aspirate a number of oocytes with a transfer pipette by applying a low negative pressure,
for example, with a rubber ball.
2. Move from well to well with the transfer pipette, carefully dropping one oocyte into each
of the wells of the well plate. Grip only the glass pipette and do not squeeze the rubber ball,
to avoid that too many oocytes will leave the pipette. The oocyte should drop into the fluid
by its weight alone.
3. If occasionally more than one cell drops into a well, this can be corrected after the plating
will have been finished.