RE: Surfactant in LSRII Sheath Buffer and Aria O-rings?

Hello Geoffrey, with regard to the LSR II we use sheath fluid with surfactant whether we
are using the HTS module or not , we still get the odd air bubble in the system but I
think this inevitable no matter what you use as a sheath fluid. I dare say it may kill
the cells but I guess this will not be an issue for you if you are not sorting	your
cells .
The question regarding the O ring for the Aria surprises me , it is our routine to remove
the nozzle from the Aria every evening and "wash off" the O ring by using plastic
distilled water bottle to gently concentrate a small stream  of water onto the ring ,
washing it into a small petri dish, we then place the lid onto the petri dish until the
next morning when we pick the O ring up with a disposable 100ul pipette tip and place the
ring onto whichever nozzle we require , it is not unusual for us to use the same O ring
for a month or longer using this daily process of removal. 
if you are leaving parts of the O ring behind it sounds to me as if you are leaving a
nozzle in the Aria for long periods and removing it when it has partialy dried out
allowing sheath crystals to form arround the nozzle and ring so when the nozzle is
removed you simply pull the ring apart due to the adhesive effects of the salt crystals
.If  you state this happens after a single time , ie one days use , then I think you need
to contact those nice people at BD



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@ncl.ac.uk
Tel 0044 191 2418831
Fax 0044 191 2418666
(mob) 0044 7970344823



-----Original Message-----
From: Geoffrey M. Lyon [mailto:geoffrey.lyon@yale.edu]
Sent: Tue 29-Apr-08 18:56
To: cyto-inbox
Subject: Surfactant in LSRII Sheath Buffer and Aria O-rings?
Hello Flowers,

I have two different questions I was hoping you all might be able to help me
with:

1) We have had quite a bit of residual air floating around our LSRIIs.	I was
told by a few different people, including a BD Service engineer, that a small
amount of surfactant in the sheath eliminates almost all the trouble we have
been experiencing with air bubbles.  Has anyone else tried this?  Does
surfactant kill most of the cells even at low concentrations since it is a
detergent?

2)  Am I the only one who now sees the Aria nozzle o-rings as a single use item?
 It seems that ever since we got the new amber style flow cell that once we take
out an o-ring that o-ring is pretty much finished.  We can actual see small
pieces of the o-ring shear off after removing the o-ring from the flow cell
after a single time.  It seems that the new flow cell is so sharp it cuts fine
pieces of the o-ring off every time it's installed.  Trying to replace a used
o-ring either leads to leaks or a pulsing stream (as part of the o-ring
flutters in and out of the stream).  We use to be able to take an o-ring in and
out of the flow cell many times before it became useless.  I've called BD and
have tried making various adjustments but it seems that we still only get a few
uses out of an o-ring.	Does anyone have a solution?

Thanks, any help is greatly appreciated.

Geoff-


Geoffrey Lyon
Biotechnology Associate I
FACS Facility
Yale University School of Medicine
TAC Building Rm-S613
300 Cedar Street

Shipping Address:
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New Haven, CT 06519

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