Re: BD HTS for LSRII/Canto

This should be a red flag that there is a problem with your  
particular setup.  We have had great luck with the HTS system...  
however, we have noticed that there is an issue in certain  
circumstances where the pressures are not quite balanced during the  
first few seconds of acquisition.  This causes the stream velocity to  
be faster until it equilibrates.  The way to diagnose this is to look  
at a graphic of a fluorescence parameter vs. time (or event number)  
-- if you see a significant change with time, you will need to gate  
out the initial few seconds of data.  The problem is only evident on  
multi-laser systems (since it is an effect on the timing), and is  
most obvious on the laser that is furthest from the first.  Also, the  
problem will be significantly exacerbated by using a small window  
extension (say, only 3); we typically run with a window extension of  
10.  (BTW, this is an issue on the LSR II; we have not seen it on the  
Calibur).

The BD instrument reps have been going over our system to trace the  
problem; they have identified a couple of possibilities and are  
working hard to correct or accommodate the issue.

By the way, I highly recommend viewing a graph of various  
fluorescence parameters vs time (or event #) on a routine basis to  
make sure that there are no clogs, or other issues that can induce  
variabilities in the fluorescence that can cause artefacts and "bad  
data."

In any case, we have 5 HTS systems, and they perform very well.   
Indeed, when we first installed an HTS unit, we discovered that our  
users wouldn't use the non-HTS-equipped system, so we were "forced"  
to buy HTS on all the machines...

mr


On Dec 8, 2006, at 2:04 PM, Watson, Susan wrote:

> We got one to use for screening mouse blood.
> After 3 months we gave up using it cos there was too
> much variation in the data as compared with hand running the samples.
> SRW
>
>
> -----Original Message-----
> From: Adrian Smith [mailto:a.smith@centenary.usyd.edu.au]
> Sent: Thu 12/7/2006 5:44 PM
> To: cyto-inbox
> Subject: BD HTS for LSRII/Canto
> Hi all,
>
> We are considering adding a 96-welll plate reader to LSRII or Canto
> and I'm putting together a grant application for the BD HTS.
>
> I'm looking for feedback from current users on reliablity and ease of
> use.
>
> I'm also particularly interested in what people are using it for?
> What sort of efficiency improvements does it provide over tubes in
> non-highthroughput scenario (ie most of what we do requires
> reasonably large files).
>
> I know I want one - I just need some convincing arguments for the
> funding committee :)
>
> Regards,
>
> Adrian Smith
> Centenary Institute, Sydney, Australia
>
>
>