labelling of Chlamydia bacteria
rafaeln at vetvir.unizh.ch
Tue Sep 19 01:43:09 EST 2000
Hi Andreas, you may consider labeling with CFSE, here is our protocol
Protocol for Staining with CFSE.
1- Cells in logarithmic growth phase are washed with PBS (x 3).
2- Resuspend them in 2 ml of PBS without FCS or protein supplement.
Final concentration: 6-10 x 107 cells/ml.
3- Add 2.8 mg/ml of CFSE.
4- Incubated cells at 37°C for 10 minutes with mixing 3 to 4 times.
5- Add several volumes of ice-cold medium to stop CFSE binding.
6- centrifugation for 10 minutes (4°C)
7- Resuspend pellet cells with fresh medium.
8- Adjust cells density to your desidered concentration
9- Optional: Add medicament(s) at variable concentrations (0 through
10- Determine the CFSE fluorescence by flow cytometry immediately after
staining, and after 24, 48, 72, and 96 hours.
I hope it will help
>does anyone have experience in permanent fluorescent labelling of live
>bacteria, in particular Chlamydia trachomatis ? We would like to quantify
>infection rates in monocytes.
>Any help would greatly be appreciated.
>Thanks in advance!
>Dr. Andreas Thiel
>phone ++49 30 28460 682
>email:thiel at drfz.de
___/\_ | Rafael Nunez mailto:rafaeln at vetvir.unizh.ch
/ o \/| | University Inst.for Virology http://www.vetvir.unizh.ch/
/ _| | Winterthurerstr. 266a Telephone: (+41) 1 6358710
/_/\__/-\/ | 8057 Zurich SWITZERLAND Faximile : (+41) 1 6358911
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