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Regarding Filler cells

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komalkumaralienboy

Contributor

Posts: 44

Joined: Tue May 20, 2014 2:24 pm

Location: Linkoping University, Sweden and Finnish Institute of Molecular Medicine, Finalnd

Post Mon Jun 01, 2015 9:44 am

Regarding Filler cells

HI,

I have few samples to run in CyTOF
but Unfortunately I lost most of the cells in washing steps this time, Thinking of Using filler cells to fill the space.
Any suggestions
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TomerWeizmann

Contributor

Posts: 43

Joined: Mon Apr 07, 2014 11:58 am

Location: Weizmann Institute of Science - Israel

Post Mon Jun 01, 2015 11:30 am

Re: Regarding Filler cells

we've mixed with irrelevant dead cells in preparation and gated out by cisplatin and markers - worked good
Dr. Tomer-Meir Salame
Head, Mass Cytometry Unit
Life Sciences Core Facilities
Weizmann Institute of Science
E-mail: tomer-meir.salame@weizmann.ac.il
https://www.weizmann.ac.il/LS_CoreFacil ... etry/about
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komalkumaralienboy

Contributor

Posts: 44

Joined: Tue May 20, 2014 2:24 pm

Location: Linkoping University, Sweden and Finnish Institute of Molecular Medicine, Finalnd

Post Mon Jun 01, 2015 12:36 pm

Re: Regarding Filler cells

Hi Tomer
what kind of cells do u use as filler cells.

I thought of using SFC9 insect cells
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Florayao

Participant

Posts: 1

Joined: Tue Jun 03, 2014 1:34 pm

Post Mon Jun 01, 2015 2:05 pm

Re: Regarding Filler cells

We added HL-60cells with the PBMCs in our limiting dilution study and found that they made no difference in our detection of cells of interest (Yao et al J. Imm.Methods 2014). We typically can get 40-60% cell recovery following staining and washing steps prior to CyTOF running. How much did you lose during your washes? Were your samples fixed well?
Yi Yao, Ph.D.
Postdoctoral Associate
Dr. Ruth Montgomery's lab
Yale University School of Medicine
Dept. Int. Med./Rheum.
300 Cedar St./TAC s420
New Haven, CT 06510
E-mail: yi.yao@yale.edu
Phone: 203-785-6955
Fax: 203-785-7053
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TomerWeizmann

Contributor

Posts: 43

Joined: Mon Apr 07, 2014 11:58 am

Location: Weizmann Institute of Science - Israel

Post Tue Jun 02, 2015 9:16 am

Re: Regarding Filler cells

we used dead splenocytes (we killed them by incubating them with perm buffer for a few seconds)
Dr. Tomer-Meir Salame
Head, Mass Cytometry Unit
Life Sciences Core Facilities
Weizmann Institute of Science
E-mail: tomer-meir.salame@weizmann.ac.il
https://www.weizmann.ac.il/LS_CoreFacil ... etry/about

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