Instrumentation |
FACS Analysis |
BD LSRII
BD LSR II ( TAC S613 )
Contact: Ewa.Menet@yale.edu or call 203-785-7958
- Multilaser multiparameter analysis cytometer
- Four lasers, 12 colors
- Digital data collection using FACS DIVA software
- Automatic compensation (compensation controls are still needed)
- 355 nm (UV): Indo-1, DAPI, Alexa Fluor 350
- 405 nm (Violet): CFP, Alexa Fluor 430, Pacific Blue, Marina Blue, Alexa Fluor 405
- 488 nm (Blue): FITC, PE, PerCP, Pe-Cy5 (aka CyChrome), PI, PerCp-Cy5.5, PE-Cy7, GFP, PE-Texas Red
- 633 nm (Red): APC, APC-Cy7, Alexa Fluor 647, Alexa 680
- Availability of UV gives a capability to perform Ca 2+ flux experiments.
- Unlike a FACSCalibur, it is possible to use PE-Cy5 and APC together.
UV Laser - 355nm 20 mW
| PMT | Bandpass | Long Pass | Fluorochrome |
| A | 530/30 | 450LP | Indo-1 (Blue), Hoechst Red, Fura-2, Am-Cyan |
| B | 440/40 | | Indo-1 (Violet), DAPI, Alexa Fluor 350, Hoechst |
Violet Laser - 405nm 50 mW
| PMT | Bandpass | Long Pass | Fluorochrome |
| A | 525/50 | 505LP | Alexa Fluor 430, CFP, Am Cyan |
| B | 440/40 | | Cascade Blue, Pacific Blue, DAPI, Hoechst, Alexa Fluor 405, BFP, Marina Blue |
Blue Laser - 488nm 50 mW
| PMT | Bandpass | Long Pass | Fluorochrome |
| A | 780/60 | 735LP | PE-Cy7, Alexa 750 PE |
| B | 670/14 | 635LP | PI (with PE), PerCP, PerCP-Cy5.5, PE-Cy5, EMA |
| C | 610/20 | 600LP | PE-Texas Red, PI (with PE and PE-Cy5.5) |
| D | 576/26 | 550LP | PE, PI (with PerCP), Pynonin |
| E | 530/30 | 505LP | FITC, GFP, CFSE, YFP, Alexa Fluor 488 |
| F | 488/10 | | SSC |
Red Laser - 640nm 40 mW
| PMT | Bandpass | Long Pass | Fluorochrome |
| A | 780/60 | 735LP | APC-Cy7, APC-Alexa Fluor 750 |
| B | 710/25 | 685LP | Alexa Fluor 680, Alexa Fluor 700 |
| C | 660/20 | | APC, Alexa Fluor 647 |
*Please consult FACS Facility Staff or http://www.bdbiosciences.com/spectra/ to determine which fluorochromes cannot be used together due to high spectral overlaps.
