[RASMB] XLA run at 37 degrees

[lelj]

Dean,

I've done runs at 30 and 40C. The main problem that I've encountered is the presence of oil in the chamber and the fact that there are two temperature controls. when you first start equilibrating you'll notice that the temperature that you read is the temperature of the 'air' in the chamber but once the vacuum goes below 100micron then you'll start reading the temperature of the rotor. So, if the rotor is at less than 37C the machine will heat up and the actual temperature in the chamber might go as high as 80C and you'll start having trouble with oil. I have the following suggestions:

1) incubate your rotor at 37C in a cell culture oven.
2)but don't incubate the cells with the sample because when you take them out of the 37C oven the windows will fog up!
3) when you start adjusting the temperature, do it step by step. Set the machine at 30C and as soon as the vacuum goes below 100 micron, look at which temperatre the rotor is and slowly increase the machine setting from that temperature, so it won't go too hot in the chamber.
4) At the end of the run before releasing the vacuum make sure to re-equilibrate the instrumet at 20C. that way you'll have less oil problem 
4)After the run, clean the windows and do a small run at 20C and then clean the windows again. you'll find that the second run after a high temperature run will have a lot of oil drops in the chamber/windows. 

good luck
barbara

PS I don't think there's a difference if it's a velocity of equilibrium run, just remember to adjust the parameters when you fit the data (like  density etc.)


-----Original Message-----

> Date: Fri May 12 09:27:39 PDT 2006
> From: "Dean Malencik" <malencid at ucs.orst.edu>
> Subject: [RASMB] XLA run at 37 degrees
> To: RASMB at server1.bbri.org
>
> Hello,
> 
> I have been asked to do a run at 37 degrees on the XLA analytical centrifuge.  Does anyone have any pitfalls to anticipate or is it even feasible to do a run at this temperature?  Any differences if the run is a velocity or equilibrium run?
> 
> Thanks.
> 
> Dean Malencik
>
--
Barbara Lelj Garolla Di Bard
Dr. Mauk's Lab
Dept. of Biochemistry and Molecular Biology
University of British Columbia
Life Sciences Building, Center for Blood Research
2350 Health Sciences Mall
Vancouver, B.C. V6T 1Z3
Phone: (604) 822-2526