Upgrading to DIVA 6.0?

Ian Dimmick ian.dimmick at newcastle.ac.uk
Wed Jan 16 17:46:07 EST 2008


Hello Michelle, I have just upgraded from diva 5 to 6 , the instruments involved were a
16 parameter 4 laser Aria and a 18 parameter 5 laser LSR II

Experiments and user database transfered without any problems , I am getting my HTS
installed later this week so can not comment on the HTS aspect.

In terms of the do's and don'ts then I would suggest the following which has worked well
for us 

Do not copy your original configuration to create customised  filter and application
specific configurations unless you have to , we felt that this complicated our use of the
instruments, in addition to a Java script error that occurred on the Aria while running
with multiple configurations, once we went back to Diva 5 then back to Diva 6 and
installed only the 1 original configuration this script error disappeared and has not
returned ,we are running very happily on both instruments with our single original
instrument configuration  derived in version 5 .

The CS&T bead function within version 6 is used complimentary to our existing QC, rather
than a replacement to it , the good & bad points are 

Laser time delays and area scaling factor calculation is very good , although on the Aria
we use this function on one pressure only (35PSI)to avoid having to set up 4 separate
configurations to accommodate each separate pressure used, we find that manual adjustment
of the other laser delays under the different pressures is easy but it would be nice to
have the ability to set all laser delays by the beads under all pressures without
creating extra configurations to do this .

Linearity check and PMT efficiency checking is excellent.

The tracking of a very comprehensive range of QC parameter results over time is
excellent.

The CS&T beads are also used to give instrument specific  minimum voltages by the method
described by Joe Trotter et al in cytometry , I have mixed feelings with regard to this ,
if the PMT is not responding in terms of efficiency or linearity then the graphical
representation that you get from the software for each PMT is very telling and is an
excellent indicator of a bad PMT, however the baseline PMT voltages we find are not very
useful , might as well just have the default 500V for each , but we deal with lots of
stem cell and "abnormal " samples so I guess everyone will have there own view on this
aspect probably proportional to types of cells under analysis.

The CS&T manuals leave a lot of questions unanswered 

We are still relatively new to version 6 so please read this mail with that in mind ,
however I guess there are not too many people in the field with a lot of experience ,
things like the ability to create a PDF of your experiment output at the click of a mouse
is a very nice touch , if I had to make a call on it I would say yes I am happy that we
went for the upgrade , and we are getting added value in terms of the core facility from
the new version .


Good luck



Ian

Ian Dimmick
Flow Cytometry Core Facility Manager
North East England Stem Cell Institute
Bioscience Centre
International Centre for life
Newcastle Upon Tyne
NE1 3BZ
UK
Ian.Dimmick at ncl.ac.uk
Tel 0044 191 2418831
Fax 0044 191 2418666
(mob) 0044 7970344823



-----Original Message-----
From: Michele Black [mailto:mblack2 at u.washington.edu]
Sent: Tue 15-Jan-08 20:29
To: cyto-inbox
Subject: Upgrading to DIVA 6.0?
Greetings,


Does anyone have first hand experience upgrading to DIVA 6.0? I am
purchasing 3 new HP workstations (xw4400's) so as far as speed, memory and
HD space I should be set. I will be putting these on my Canto, LSRII and
Aria. 


Questions

1. Can the experiments and user database be transferred to 6.0 from a
previous version from an old workstation? 

2. Anyone using HTS on an LSR II, do the HTS templates transfer OK?

3. Do's and don'ts when upgrading? If you have already gone through this and
know of some things that will save me from pulling my hair out I would love
to hear what you have to say.

4. General impression of the improvements from the previous version?


Cheers,


Michele


Michele Black 
Director | Cell Analysis Facility 

Department of Immunology

University of Washington



Lab:

(206) 685-3014 


Fax:

(206) 543-1013 

 <mailto:mblack2 at washington.edu> mblack2 at washington.edu 








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