replies to questions about 96 plate reader for LSRII

I have made a compilation of the relevant replies to my inquires 
about the plate reader-Please contact me directly if you would like 
the respondents emails-I also received replies about the HTS for the 
Calibur but did not include those here.

Thanks so much to everone who responded-I think it will help alot.

Janet


I have been using the LSRII for 2.5 years now and we just had the HTS
installed about 3 months ago.  Our users love the LSRII and now they
love the HTS.  We had a few problems with pressure build up but changed
the filter on the waste container and that seemed to take care of the
problem.  The software is easy to use (obviously it could be better but
it gets the job done).	The users are happy with the performance and
reliability.  One lab has been screening hybridomas on the HTS and it
has saved them hours.  Instead of 150 tubes its two plates in about 30
min.  The HTS primes during start up and then mixes and rinses in
between samples to minimize carryover.
The one complaint that I hear a lot is that when users want to run tubes
the HTS is in the way.	I don't take off the HTS because it would be too
much work to keep changing it out.  There is enough room to reach over
the unit and put on a tube.  Bottom line we are satisfied with the
product.


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We have a LSRII with a HTS (High Throughput Sampler) here at UCSF/SFGH. We
haven't done much on it, but it has been running fine until recently. We
noticed and suspects that with a high concentration of cells (~6mil PBMC /
200ul), the in between wash from well to well isn't enough to clean the
tubing and cells are getting carried over. We are underway in determining
the maximum concentration and hope to solve this issue soon. Also, HTS only
takes shallow well plates only.

Hope this helps.





  the HTS is running fine and reliably on the LSR II. Its operation is 
integrated into Diva now. I had an early version of it at my previous 
work place that had certain shortcomings but not much. It is so good 
I am buying three of them right now for LSR II cytometers.

Call me if you need more info.






Chris Trussell at GNF ( <ctrussel@gnf.org>) has one and I just talked 
to him yesterday and he said it was great (I have it for the Calibur, 
and am considering one for the LSR IIs myself).  Email him for a more 
candid opinion






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we have a LSRII with the plate reader attachment (which incidentaly can also
read 384 well plates), and I have found it to be extremely reliable.

The paragraph below is cut and pasted from an email from a BD consultant

User selectable 96 and 384 well microplate formats.
Sample delivery rate and volume controlled via precision syringe pump.
User selectable acquire volume (2-200 uL) and rate (0.5 - 3.0 uL/sec)
FAST - less than 15 min per 96 well plate in "high throughput mode".
removes only 22 uL from the well, for up to 10 uL acquire volume.
User selectable mixing and wash parameters.

I routinely process somewhere between 120 and 200 samples in a session. With
the attachment I am able to process two trays in about an hour (compared to
about 4 hours with tubes). However, the speed is very much dependent on sample
concentration. I work with primary cells from popliteal nodes of mice so I
don't have large cell numbers. Another person in our lab works with cell lines
and has achieved an acquisition rate similar to that outlined in the
specifications given above.

If cell numbers are not an issue, then you will find the speed and "hands
free" nature of the attchment to be fantastic. If you are looking at analysing
rare event samples then the only advantage of the plate attchment is
automation.

Overall I am very happy with the attachment itself, but unfortunately the part
of the software driving it is not as well designed as the rest of the DiVa
software. However these difficulties have not been serious enough for me to
consider discontinue using the attachment.

Furthermore, if you wish to continue using the machine to analyse tubes as
well as plates be prepared to spend around 20 minutes converting the machine
over from plate format to tube format, and about 5 minutes from tubes to
plates. It takes longer to revert to plate format due to the necessity of
flushing the plate reader with MQ water if it will not be used for more than 7
days. There is no cleaning program written for this procedure, you have to
manually prime it 10 times.

If you would like more specific information let me know, I would be more than
happy to help.



Here is advantages and disadvantages of High Throughput Sampler(HTS).

Advantages
1. Easy to install or removal
2. Fast sample acquisition
3. Easy maintenance and cleaning

Disadvantages
1. No HV adjusting between the samples
2. Need higher concentration of cells because the limitation of volume

Even if I found a couple of disadvantages, HTS works fine.
Hope it helps,


-- 
Janet Dow
Laboratory Research Specialist and Manager
Flow Cytometry and Cell Sorting  Facility
North Carolina State College of Veterinary Medicine
4700 Hillsborough Street
Room C-309
Raleigh, NC 27606
(919)513-6443