[Cytometry] Sorting neurons

Daqian Sun daqian.sun11 at gmail.com
Mon Apr 22 16:08:57 EDT 2013

Hi All,
   We have a user who want to sort neurons isolated from mouse tissue. We
have a few technical questions:
1. There are tons of dead cells and debris in his sample. I wonder of
anyone have a good protocol for preparation of neuron cell suspension.
2. Currently we are using AriaII, 100um nozzle and 20psi for his sort. Do
we need a larger nozzle and lower pressure?
3. There is a literature which suggest to use low flow rate, 1000-3000
cells per seconds for neuron sorting. I'm assuming that the neurons also do
not like high differential pressure, is this right?
4. The FSC-W and SSC-W signal distribution are much wider than other type
of cells. The user told me that the neuron he isolated are not round in
suspension. Does anyone has experience sorting irregular shaped cells? Can
we still use signal width for doublet discrimination?

Dr. Daqian Sun
Senior Scientist, Manager
Mount Sinai School of Medicine
Flow Cytometry Shared Resource Facility New York, NY  10029

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