 | Make appointment at least two days early and discuss
with Randy first about your project. Make sure you have the cell show the
levels of expression that are to be sorted. You welcome to use Coulter XL to
run the sample first see how the expression. |
| Preparing the sample, resuspend them in your culture
medium that contains HEPES buffer. It will protect the cells from any PH
changes they might undergo during pressurization or charging of droplets
during the sort process. |
| For the live cell sorting, the buffer should contain
1% BSA or 1% Fetal Calf Serum. |
| The concentration should be around 10-20 x 106
cells/ml. |
| Tube should use 12x75mm, 5ml Polypropylene Round-Bottom
Tube. (Falcon#352005) |
| Filter the cells before sorting. Use cell strainer 70um
(Falcon#352350) |
| Collecting sorting sample: Variety of containers.
Common one is 12x75mm falcon tube. Or 15 ml conical tube. Or any size of
microtiter wells plate. 96 wells is the most popular one. |
| The entire collect container should have your culture
medium. You may want to put double dose of fetal calf serum or BSA. Make your
cells a little happier. |
