RE: Do your have any DIVA problems?

To all,

We are routinely collecting 2.5M cell counts on most of out multicolor work.
The software works (DiVa.v4.0.1) quite well with only a few limitations as
listed below, which BDIS is working to improve. With the additional
detectors (20 parameters) the use of DAQ II boards is required.

1. Routinely files should be transmitted and stored onto larger file servers
and deleted from the DiVa hard drive.

2. Routinely the computer requires defragmentation with the DEFRAG program
located in the accessories folder.

3. If files do not reach predetermined target counts these files slow down
tube runs because the graphics are re-loaded, slowing down memory
allocation. However, if these counts are reached then re-graphing is
seamless and memory allocation can proceed without slowing down the system.
Many tubes per experiment can be collected without difficulty. Hence, we
would like researchers to "hit" their pre-determined counts, which is not
always possible. In these cases one can duplicate the experiment and
continue collection in this new file.

4. We found that the "NEXT" button should not be used. At least in our
system it will lock-up the software without warning. Tubes can be selected
and advanced by selecting the pointers located beside each tube.

Hope this helps.

SP

Stephen P. Perfetto, MS.,MT. (ASCP)Manager, Core Flow Cytometry FacilityVaccine
Research Center, NIH40 Convent Dr., Room 5507Bethesda, MD 20892-3015

> ----------
> From: 	Fischer, Randy (NIH/NIAMS)
> Sent: 	Tuesday, October 19, 2004 9:56 AM
> To:	Cytometry Mailing List
> Subject:	RE: Do your have any DIVA problems?
> 
> Obviously I missed this when it first came out, but is it true the LSRII
> with the DiVa is limited to collecting 100K events/tube?  Mario and Steve,
> you guys are doing >14 colors per tube, and statistically, small
> populations
> could not be acquired in enough numbers with this limitation. As you two
> provide the impetus for the rest of us to push the frontiers with huge
> polychomatic experiments, how does this work? Or do you just limit your
> experiments to a few tubes?
> 
> I do find it hard to believe BD would not have addressed this problem
> before
> now, it was a problem with the original DiVa unit we had (gone now).
> 
> Randy T. Fischer
> NIH/NIAMS
> Building 10, Room 6D50
> 9000 Rockville Pike
> Bethesda, MD 20892
> (301) 594-3537
> fischer1@mail.nih.gov
> 
> > ----------
> > From:	Christopher A. Worth
> > Sent:	Friday, October 15, 2004 4:12 PM
> > To: cytometry@flowcyt.cyto.purdue.edu
> > Subject:	RE: Do your have any DIVA problems?
> > 
> > 
> > That sounds like a plan.
> > 
> > Customer 'our cytometer that you built isn't working right with our
> > experiments"
> > 
> > Vendor 'your experiments aren't working right with OUR cytometer.'
> > 
> > Customer 'hmmmm.... what to do? '
> > 
> > Vendor 'change your experiments!'
> > 
> > Customer 'or I could change vendors'
> > 
> > sounds simple enough to me.
> > 
> > Chris
> > 
> > 
> > 
> > 
> > --Original Message Text---
> > From: Akos Szilvasi
> > Date: Fri, 15 Oct 2004 10:12:39 -0400
> > 
> > 
> > I pasted below a response to our Diva problems. 
> > 
> > In short there are some serious limitations "built in" to Diva that
> > renders the LSR II to a less than perfect cytometer. 100,000 event limit
> > is unacceptable for any cytometer. Especially not for one that can
> collect
> > over a dozen parameters. If you apply complex gating to your analyses,
> run
> > an Indo-1 Ca++ flux experiment or go after rare events you may need more
> > than one or two hundred thousand cells collected. The recommended 50
> > tube/experiment is a new feature I have not seen in the color catalog I
> > received. I guess the delay of the HTS's release for LSR II is due to a
> > shortage of 50 well Falcon plates. 
> > 
> > What is the solution? We are very interested in BD's official response.
> We
> > are about to buy three more of the new BD cytometers. 
> > 
> > Akos 
> > 
> > .............................................................. 
> > 
> > As a general rule, when collecting 10 parameters per Tube, approximately
> > 50
> > Tubes per Experiment can be recorded with 100,000 events per Tube.
> > Collecting more parameters (such as Area and Height) or having multiple
> > gates decreases the number of Tubes to 25 or fewer per Experiment. To
> save
> > memory (and disk space for permanent storage), save only parameters that
> > are being used.
> > 
> > . once they notice that Diva is slowing down,
> > that's a sign that their open Experiment is getting too large and they
> > should split it up into smaller experiments. They should also check the
> > size of their database and clean it out on a regular basis.
> > 
> > 
> > 
> > 
> > 
> 
>