Isolating cells from Haliclona

[Gary Laevsky]

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Hi All,

We have a group trying to isolate cells from a Haliclona sponge.

When they homogenize, they get cells, bacteria, and various other particulates.

Then, when they try to sort, the cells explode.

The cells are dead, so no adherence and no internalization of magnetic particles.

They've tried centrifugation once, with no success. 

Thinking about trying to capture with a microinjector, gentle vacuum, and then isolating, but I think cell will get torn up pulling out of media.

Maybe  a filter?

Other ideas?

Thanks in advance.

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[JOEL B. SHEFFIELD]

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How are you treating the tissue before homogenization?  Wasn't Haliclona one of the species that was used by Moscona and Humphreys to show species specific reaggregation? 

On Tue, Sep 6, 2016 at 12:48 PM, Gary Laevsky <[hidden email]> wrote:

***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. *****

Hi All,

We have a group trying to isolate cells from a Haliclona sponge.

When they homogenize, they get cells, bacteria, and various other particulates.

Then, when they try to sort, the cells explode.

The cells are dead, so no adherence and no internalization of magnetic particles.

They've tried centrifugation once, with no success. 

Thinking about trying to capture with a microinjector, gentle vacuum, and then isolating, but I think cell will get torn up pulling out of media.

Maybe  a filter?

Other ideas?

Thanks in advance.

--