Equipment
Our lab has a FacScan three color one laser cell analyzer with a
G3 computer, a FacsCalibur four color two laser cell analyzer with a G5 computer, and a LSRII
10 color two laser cell analyzer. We also have a MoFlo seven color three laser cell sorter and
a FacsAria 13 color three laser cell sorter. We have a G4 with FlowJo and Cellquest software
and a PC analysis computer with Diva and Summit.
Equipment Specifications:
FACScan
Using a G3 computer and Cellquest Ver. 3.5
| |
Fluorochromes |
|
|
| |
|
|
|
| 488 nm: |
|
|
FILTER(nm) |
| 1 |
FITC/GFP |
|
530/30 |
| 2 |
PE/CY3 |
|
585/42 |
| 3 |
PE-TXRD/PI/7-AAD |
|
650LP |
| |
PE-CY7/PerCP/PE-CY5 |
|
|
FACSCalibur
Using a G5 computer and Cellquest Ver. 5.2
| |
Fluorochromes |
|
|
| |
|
|
|
| 488 nm: |
|
|
FILTER(nm) |
| 1 |
FITC/GFP |
|
530/30 |
| 2 |
PE/CY3 |
|
585/42 |
| 3 |
PE-TXRD/PI/7-AAD |
|
670LP |
| |
PE-CY7/PerCP/PE-CY5/PerCP-CY5.5 |
|
|
| |
|
|
|
| 633 nm: |
|
|
|
|
1 |
APC/CY5 |
|
661/16 |
MOFLO
Using Summit software Ver. 4.3
| |
Fluorochromes |
|
|
| |
|
|
|
| 488 nm: |
|
|
FILTER(nm) |
| 1 |
FITC/GFP |
|
530/40 |
| 2 |
PE/CY3/YFP |
|
580/30 |
| 3 |
PE-TXRD/PI/7-AAD |
|
630/30 |
|
4 |
PE-CY5/PI/7-ADD/PerCP |
|
670/30 |
|
5 |
PE-CY7 |
|
740LP |
| |
|
|
|
| 633 nm: |
|
|
|
|
1 |
APC/CY5 |
|
670/30 |
|
Can use one 2 |
APC-CY5.5/APC-CY7 |
|
700ALP |
|
or the other 2 |
TEXAS RED |
|
630/30 |
| |
|
|
|
| 354 nm: |
|
|
|
|
1 |
SEE CHART BELOW |
|
|
| |
|
|
|
|
UV Dyes |
for MOFLO |
|
|
| |
|
ex nm |
em nm |
|
Hoechst 33342 |
|
350 |
470 |
|
DAPI |
|
359 |
462 |
|
Cascade Blue |
|
375/400 |
423 |
|
EBFB |
|
380 |
440 |
|
Marine Blue |
|
365 |
460 |
FACSAria/LSR II
Using DIVA software Ver. 4.1
| |
Fluorochromes |
|
|
| |
|
|
|
| 488 nm: |
|
|
FILTER(nm) |
| 1 |
GFP |
|
510/21 |
| 2 |
FITC/YFP |
|
530/30 |
| 3 |
PE |
|
576/26 |
| 4 |
PE-TXRD/PI/7-AAD |
|
610/20 |
| 5 |
PE-CY5/PI/7-ADD/PerCP |
|
660/20 |
| 6 |
PE-CY6.6/PerCP-CY5.5 |
|
695/40 |
|
7 |
PE-CY7 |
|
780/60 |
|
|
|
|
|
|
633 nm: |
|
|
|
|
1 |
APC/CY5 |
|
660/20 |
|
2 |
APC/CY5.5/ALEXA 700 |
|
700/30 |
|
3 |
APC-CY7 |
|
780/60 |
|
|
|
|
|
|
407 nm: |
Aria only |
|
|
|
1 |
Alexa Fluor 430 |
|
530/30 |
|
2 |
Pacific Blue/EBFB/ECFP |
|
450/40 |
|
3 |
Alexa 405/Cascade Blue |
|
425/10 |
|
|
|
|
|
|
LSRII will have same capabilities as the FacsAria except for the 407nm Laser. |
|
|
|
For analysis we have a G4 with Cellquest Ver. 5.2 and FlowJo Ver. 8.3 and a PC with
Summit Ver. 4.3 and DIVA Ver 4.1. |
CYAN
Using Summit Software Ver. 4.3
| |
Fluorochromes |
|
|
| |
|
|
|
| 488 nm: |
|
|
FILTER(nm) |
| FL1 |
FITC/GFP |
|
530/40 |
| FL2 |
PE/YFP |
|
575/25 |
| FL3 |
PE-TXRD/PI/7-AAD |
|
613/20 |
| FL4 |
PE-CY5/PI/PerCP |
|
680/30 |
| FL5 |
PE-CY7/PE-Cy5.5 |
|
750LP |
|
|
|
|
|
|
405 nm: |
|
|
|
|
FL6 |
CASCADE BLUE/CFP/DAPI |
|
450/50 |
|
FL7 |
CASCADE YELLOW |
|
530/40 |
|
|
|
|
|
|
635 nm: |
|
|
|
|
FL8 |
APC/CY5 |
|
665/20 |
|
FL9 |
APC-CY5.5/APC-CY7 |
|
750LP |
|
|
|
|
|
|
