FRET using FACScalibur
szollo at jaguar.unideb.hu
Tue Feb 15 15:16:06 EST 2005
The paper bleow describes how to perform FRET measurements on
FACScalibur. It also offers autofluorescence correction on a cell by cell
Sebestyén, Z., Nagy, P., Horváth, G., Vámosi, G., Debets, R., Gratama, J.
W., Alexander, D.R., Szöllösi, J: Improving flow cytometric energy transfer
measurements on a commercial flow cytometer using cell-by-cell correction
Cytometry, 2002. 48, 124-135.
However in your case the situation can be a bit more complicated. Can I
assume from your short description that the ligands and the receptors are
on two separate cells? If yes, how would you keep the cells together in
the flow system? Even if you succeed only percent of the receptors will
interact with the ligands [for sterical reasons] so the FRET signal will be
overshadowed by fluorescence coming from molecules not participating in the
For this purpose microscopic measurements could be a better solution.
Do not hesitate to write me directly with more specific questions!
At 11:04 PM 2/11/2005, Garg, Ankita wrote:
>I have to perform receptor-ligand interaction on 2 cell .I want do it
>using FRET, but am
>not sure how can I do it with flowcytometr . I work on FACScalibur with 2
>help me out.
>thanks in advance
>From: Nebe-Von-Caron, G [mailto:g.nebe-von-caron at unipath.com]
>Sent: Thursday, February 10, 2005 12:40 PM
>Subject: RE: Sorter sterilization
>According to a beer advert there are beers that reach parts not reached
>by others, but I can not see that to be the case for disinfectants.
>Hypochlorite and alcohol is as potent as you can get. You are more
>likely to miss some crucial part in the cleaning routine (valve, air
>trap...). If you sort below 40PSI I recommend Millifill-GV disposable
>online filter(2 in parallel) close to the flow cell. That should stop
>anything from that end.
>From: Barren, Phil [mailto:BarrenP at MedImmune.com]
>Sent: 04 February 2005 21:35
>Subject: Sorter sterilization
>Can someone tell me the concentration of hydrogen peroxide they use in
>sterilization of a cell sorter.
> I have tried ETOH and Bleach ( various concentrations), but my low
>level contamination continues, so I want to try peroxide.
>Any wisdom would be appreciated
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