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>Does anyone have any experience using HBSS (Hanks Buffered Salt Solution)
>as sheath in a sorter? We've been using it for some sorts over the last
>couple of months because one of our users feels it increases cell viability.
>We have problems phasing the side streams and it clogs our sheath filter.
>Our usual buffer is PBS w/o Ca++ and Mg++ and we have no problems with it.
We also have tried HBSS as sheath and had nozzle problems. In our case, it
seemed to be due to HEPES buffer in the HBSS and leaving that out helped the
problem. Also, some of the components of HBSS can precipitate and cause
problems. Unless you are sorting large numbers of cells so the volume in the
collection tube has a substantial contribution from the sheath, your user
should be able to provide HBSS (or whatever the cells like) as a medium for the
collection tube. The time spent in the sheath is minimal and it should not
matter what the composition is. We usually include at least 10% serum in the
collection tube, this increases viability of sorted cells. Some people even
sort into 100% serum thinking that more is better. I have certainly found that
viabilities of primary isolate cell populations (compared to cultured cell
lines) is much better if the collection tube contains serum or some other
source of protein. Cooling the collection tube is also helpful. But my opinion
is that the sheath fluid should not really matter unless that is the primary
component that the cells collect in after sorting.
Cheers,
Jerry Spangrude
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