RE: PI viability dye during sorting

Art:

Yes, it is called aerosol containment devices, no sort operator should be
caught without it. If your instrument doesn't have one then you should be
seriously consider using a Personal Protective device. What do you do when
you sort other live material? I think I would be more concerned about
aerosol inhalation of other biohazardous materials from live cells. Kevin
Holmes did a very nice evaluation of aerosol generation from an Aria cell
sorter characterizing both size and quantity of aerosols generated under a
variety of conditions and showed a good percentage are just the right size
for inhalation into the lung. He presented this at ISAC and will be
presenting it at the upcoming Chesapeake Cytometry meeting next Monday in
Rockville, MD. Sorting without some type of aerosol containment practice is
at best risky.


Joanne Lannigan, M.S.

Director, Flow Cytometry Core Facility

University of Virginia

Jordan Hall, Room 7067

P.O. Box 800734

Charlottesville, VA 22908-0734

Office: 434-924-0274

Lab: 434-243-2695 or 434-243-2711

Fax: 434-982-1071

email: joannelannigan@virginia.edu 


  _____  

From: Arthur Roberts [mailto:robertar@umdnj.edu] 
Sent: Tuesday, May 27, 2008 12:20 PM
To: cyto-inbox
Subject: PI viability dye during sorting


Hi Mario,
I saw your answer to this question, and I was wondering about the risks in
using PI in a stream-in-air sorter. Since PI is a DNA intercalater, and
therefore, could have nasty effects on the operator's DNA if it were inhaled
as an aerosol, is there a way to use it in sorting that will minimize
exposure?  I would wash the cells before sorting, but the PI intensity drops
off quickly after washing, since the binding is reversible.  

Thanks for any insight you might have.

Art

Arthur Roberts
Robert Wood Johnson Medical School
Dept. of Molec. Genetics, Microbiol & Immunol
SRB rm 113
675 Hoes Lane
Piscataway, NJ 08854
phone: 732-235-4502
fax:   732-235-4505


----- Original Message -----
From: Mario Roederer <roederer@drmr.com>
Date: Sunday, May 25, 2008 4:24 pm
Subject: Re: Amine-reactive dyes and impaired functionality
To: cyto-inbox

> We have not looked at the effect of amine-reactive dyes on long-
> term	
> viability.  However... why use the amine-reactive dyes if 
> you want to 
> culture long-term?  The amine reactive dyes are good 
> because they	
> withstand fixation.  If you're not going to fix, you should 
> use PI,  
> the king of all viability dyes!
> 
> mr
> 
> On May 23, 2008, at 9:43 PM, Suzanne Mertens wrote:
> 
> > Dear Colleagues,
> > I have a Core client who is interested in using amine-
> reactive  
> > Alexa430 as a viability
> > dye during a cell sort. Should I be  concerned about 
> impaired  
> > functionality for the
> > sorted cells previously stained with such a reagent?. Is there 
> any  
> > data out there for
> > functional performance of cells stained with an amine reative 
> dye,	
> > sorted, and then
> > cultured for cytokine assays?
> >
> > Suzanne
> >
> > Sent from my Verizon Wireless BlackBerry
>