[Cytometry] Clogging from running digested mouse lungs

Ryan Rountree Ryan.Rountree at bn-immunotherapeutics.com
Tue Feb 3 13:31:20 EST 2009


Hi All, 

 

We have been getting some clogs in our FACScalibur after running
digested lung samples. The lungs are prepped by mincing and digesting
with DNAse/Collagenase. Then squishing and straining through 70um nylon
filters. There is lots of debris in the prep when you look under the
microscope, and the pellets for 1E6 cells are much larger than 1E6
splenocytes. But I don't think we're having any problem with
resuspending the cells. Others on this site are using a similar
protocol.

 

However, the day after running our samples, people have found clogs in
the SIT. I'm going to start checking the SIT for buildup routinely now,
but we think these samples are causing the problems. We also run a lot
of whole blood, which tends to be viscous and could cause build-up over
time.

 

Any advice for how to clean better??

 

This protocol is run for cleaning after our session:

 

5 min 10% Contrad 70 on hi

1 min 10% C-70 with the arm to the side

1 min 10% bleach with the arm to the side

5 min 10% bleach on hi

1 min water with the arm to the side

5 min water on hi

 

Leave tube of water on the SIP

 

 

Thanks for your help,

Ryan

Ryan Rountree, Ph. D. 
Research Scientist II
BN ImmunoTherapeutics, Inc.
2425 Garcia Ave
Mountain View, CA 94043
Desk: 650-681-4673
Fax: 650-681-4680

 



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