[Cytometry] Core facility questions

Charles A Kuszynski ckuszyns at unmc.edu
Thu Feb 25 11:29:01 EST 2010

I think Grace has a good suggestion in attending the Core Managers Workshop at Cyto 2010.  This is a good forum to get the most feedback.  It's coming up soon,
As for your questions, we have found that it usually takes 10-30 minutes to change nozzles on the Aria I.  However, once you have done this, you should have written down or saved somewhere what the optimal settings for each nozzle and cell type.  In our hands, doing this greatly reduces the time required to change setup pressure and nozzle size.
As for the other issues, we like Grace charge for the actual time the instrument is being used by each investigator.  This includes setup time and data handling.  We too have more than one sorter.  The Aria II is much easier to change nozzles and pressures.  But again keeping track of the "Sweet Spots" in terms of nozzle size and frequency etc. will make these switches easier and faster.
The suggestion of scheduling similar experiments on the same instrument has merit, although we have found that this is often not possible as our sorts vary from day to day.  We sort bacteria, blood from numerous sources and cell lines and tumor homogenates all in the same day.  We have experienced little contamination or loss of cells due to carry over or sample contamination as long as you observe good lab practices such as disinfecting the area around the sorter and thoroughly flushing the instrument.  Remember (in theory) the only place you should have sample is in the sample pick-up and in the core stream as well as potentially on the sort plates and sort chamber.  Cells should not accumulate in the flow cell as the core stream is significantly smaller than the core or the nozzle.
See you at the core managers meeting in Seattle.

Charles A. Kuszynski, Ph.D.
Associate Professor
Director, Cell Analysis Facility
University of Nebraska Medical Center
985816 Nebraska Medical Center
Omaha, NE 68198-5816
402 559-6299 office
402 559-6267 lab
402 980-7654 cell
402 559-4069 fax
ckuszyns at unmc.edu

"What a long strange trip it's been" Grateful Dead

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-----cytometry-bounces at lists.purdue.edu wrote: -----

To: "cytometry at lists.purdue.edu" <cytometry at lists.purdue.edu>
From: Mok Michelle <michelle_mok at bsf.a-star.edu.sg>
Sent by: cytometry-bounces at lists.purdue.edu
Date: 02/24/2010 07:25PM
Subject: [Cytometry] Core facility questions

Dear Flow-ers,

I have questions from 1 facility operator to another. I'm just wondering how other core facilities are handling situations like these:

1)      In a core facility, we accept many types of cells for sorting. At our lab, we have received everything from beads to primary tissue to tissue culture cells... The outcome of each sort can depend greatly on the type and size of cell passing through the machine. How do you offer service to ensure that we are giving our users the most optimal sort? Do you do an optimization procedure for each type of cell user brings down or is there like "fail-safe" setting for all/most cells.

2)      Do you "pad" bookings between users, like perhaps set ½ hr between users to do run a FACSRinse or Backflush?

3)      At our facility, we have the FACSAria 1, everytime we need to change between different sort setups (e.g. Low to High), it means a down time of about ½ hour, to change the nozzle and re-stabilize the stream. Is that normal? How often do you change the sort setup within a single day? If (as a core facility) we have to serve as many users as possible, how are other facilities doing it to decrease the downtime required?

At our facility we have 2 FACSAria (of different specs). What possibilities could we have to deal with such situations?

Advance thanks to anyone who is willing to share.

Best Regards,
Michelle Mok
Manager | Scientific Services | Biopolis Shared Facilities
TEL +65 6407 0148 (Office) | FAX +65 6478 9924
michelle_mok at bsf.a-star.edu.sg<mailto:michelle_mok at bsf.a-star.edu.sg>

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