Instrumentation
Cell Sorting Instruments | Analysis Instruments |
FACSAriaII (Hill 480) | LSRII 14-Color (CRB 267) |
MoFlo (Hill 480) | LSRII 18-Color (Hill 481) |
| CyAn ADP (Hill 480) |
| FACSCalibur (Hill 481) |
| EPICS XL-MCL (Hill 480) |
Sorting Instruments
FACSAriaII 16-color cell sorter, BD (purchased 2009, Violet laser upgrade 2013) (Hill 480)
● Operated by facility personnel only
● Four lasers; Blue (488nM), Red (640nM), Violet (405nM), UV (325nM)
● Capable of sorting up to 4 subpopulations simultaneously at rates up to 21,000 cells/second (70u) or 9,000 cells/sec (100u)
Suggested fluorochromes (there are many more – see spectra viewers):
Blue Laser (488nM) x 5 parameters | FITC / GFP / Alexa488 (530/30) PE / DsRED / PyroninY (575/25) PE-TxRED / PI / mCherry (610/20) PerCP-Cy5.5 / PE-Cy5 or 5.5 / 7AAD (710/50) PE-Cy7 (780/60) |
Red Laser (640nM) x 3 parameters | APC / Alexa647 (670/30) Alexa 700 (710/50) APC-Cy7 / Alexa 750 or 780 (780/60) |
Violet Laser (405nM) x 6 parameters | PacBlue/BV 421/CellTrace Violet (450/50) AmCyan/Aqua Live-Dead/BV 510 (510/50) BV 570/Qdot 585 (585/30) BV 605/Qdot 605 (610/20) BV 650/Qdot 655 (660/20) BV 685/Qdot 705 (710/50) |
UV Laser (325nM) x 2 parameters | Hoechst Red / Qdot 655 (670/50) Hoechst Blue / DAPI / BV 421 (450/50) |
Numbers in parentheses refer to emission filter wavelengths of each detector
Contact facility with questions regarding color combinations.
MoFlo 6-color Sorter, DakoCytomation (purchased 2001) (Hill 480)
● Operated by facility personnel only
● Two lasers; Blue (488nM), Red (635nM)
● Capable of sorting to 4 subpopulations simultaneously at rates up to 25,000 cells/second (70u) or 10,000 cells/sec (100u)
Suggested fluorochromes (there are many more – see spectra viewers):
Blue Laser (488nM) x 4 parameters | FITC/GFP PE PE-TxRed or PI PerCP(Cy5.5)/PE-CY5(5.5)/ 7AAD/PE-Cy7 |
Red Laser (635nM) x 2 parameters | APC Alexa 700 or APC-Cy7/Alexa 750/780 |
Numbers in parentheses refer to emission filter wavelengths of each detector
Contact facility for other fluorochromes not listed, or with questions regarding color combinations.
Analysis Instruments
LSRII (14-color), Becton-Dickinson (purchased 2007) (CRB 267)
● Capable of user-friendly analysis
● Four lasers; Blue (488nM), Violet (405nM), Red (635nM), Green (532mM)
● High-speed analysis-only instrument
Suggested fluorochromes (there are many more – see spectra viewers):
Blue Laser (488nM) x 3 parameters | FITC / GFP / Alexa488 / YFP (530/30) PE_Blue (575/26) PerCP-Cy5.5 / PI (695/40) |
Violet Laser (405nM) x 3 parameters | AmCyan / Aqua Live/Dead (525/50) PacBlue (450/50) Qdot 605 or 655 (670/14 or 610/20)* |
Red Laser (635nM) x 3 parameters | APC / Alexa647 (660/20) Alexa700 / APC-Cy5.5 (710/50) APC-Cy7 / Alexa750 or 780 (780/60) |
Green Laser (532nM) x 5 parameters | PE-Gr / dsRed (585/42) PE-TxRed / dsRed / mCherry (610/20) PE-Cy5 / PerCP (670/30) PE-Cy5.5 (730/45) PE-Cy7 (780/60) |
Numbers in parentheses refer to emission filter wavelengths of each detector
*Contact facility personnel for instructions
Contact facility with questions regarding color combinations.
LSRII (18-color), Becton-Dickinson (purchased 2008) (Hill 481)
●Capable of user-friendly analysis
●Four lasers; Blue (488nM), Violet (405nM), Red (640nM), Green (532mM)
●High-speed analysis-only instrument
Suggested fluorochromes (there are many more – see spectra viewers):
Blue Laser (488nM) x 2 parameters | FITC / GFP / Alexa488 / YFP (525/50) PerCP-Cy5.5 / PI (710/50) |
Violet Laser (405nM) x 8 parameters | PacBlue BV421 (450/50) AmCyan / Aqua Live/Dead / BV510 (525/50) Qdot 565 BV570 (560/40) Qdot585 (585/42) Qdot 605 or 625 / BV605 (605/40) Qdot 655 / BV650 (660/40) Qdot 705 / BV711 (705/70) Qdot 800 / BV785 (780/60) |
Red Laser (640nM) x 3 parameters | APC / Alexa647 (660/20) Alexa700 / APC-Cy5.5 (710/50) APC-Cy7 / Alexa750 or 780 (780/60) |
Green Laser (532nM) x 5 parameters | PE-Gr / dsRed (575/25) PE-TxRed / dsRed / mCherry (610/20) PE-Cy5 / PerCP (660/20) PE-Cy5.5 (710/50) PE-Cy7 (780/40) |
Numbers in parentheses refer to emission filter wavelengths of each detector
Contact facility with questions regarding color combinations
CyAn ADP, DakoCytomation (purchased 2002) (Hill 480)
● Capable of user-friendly analysis
● Four lasers; Blue (488nM), Red (635nM), Violet (405nM)
● High-speed analysis-only instrument
Suggested fluorochromes (there are many more – see spectra viewers):
Blue Laser (488nM) x 5 parameters | FITC / GFP / Alexa488 PE / DsRED PE-TxRED / PI / mCherry PerCP-Cy5.5 / PE-Cy5 or 5.5 / 7AAD PE-Cy7 |
Red Laser (635nM) x 2 parameters | APC / Alexa647 Alexa 700 |
Violet Laser (405nM) x 2 parameters | AmCyan / Aqua Live/Dead PacBlue /BV421 |
Contact facility for other fluorochromes not listed, or with questions regarding color combinations.
FACSCalibur, Becton-Dickinson (purchased 2004) (Hill 481)
●Capable of user-friendly analysis
●Two lasers; Blue (488nM), Red (635nM)
●Low-speed (up to 5,000 events per second) analysis-only instrument
Suggested fluorochromes:
Blue Laser (488nM) x 3 parameters | FITC / GFP / Alexa488 / YFP PE / DsRED PE-TxRED / PI or 7AAD / mCherry / PerCP-Cy5.5 / PE-Cy5 or 5.5 / PE-Cy7 |
Red Laser (635nM) x 2 parameters | APC / Alexa647 / Alexa 700 or 750 or 780 |
Contact facility for other fluorochromes not listed, or with questions regarding color combinations.
EPICS XL-MCL, BeckmanCoulter (purchased 1994) (Hill 480)
● Capable of investigator-operated analysis
● Single Blue laser (488nM)
● Low-speed (up to 5,000 events per second) analysis-only instrument
● MCL autoloader carousel allows for walk-away operation
Suggested fluorochromes (there are many more – see spectra viewers):
Blue Laser (488nM) x 4 parameters | FITC / GFP / Alexa488 / YFP PE / DsRED PE-TxRED / PI / mCherry PerCP-Cy5.5 / PE-Cy5 or 5.5 / 7AAD / PE-Cy7 |
Contact facility for other fluorochromes not listed, or with questions regarding color combinations.
Other related equipment available in Hill 480
Fluorescent microscope
Additional PC and Mac workstations for post-acquisition analysis
FlowJo post-acquisition software
Refrigerator
There is also a collection of the following publications and resources:
● Current Protocols in Cytometry
● Cytometry, The Journal of the International Society for Analytical Cytology (1988 – current)
● Various flow cytometry reference books, including Practical Flow Cytometry (Shapiro, both 2nd & 3rd editions), Handbook of Flow Cytometry Methods (Robinson), and more
● Catalogs of flow cytometry-related supplies, antibodies, fluorochromes, etc.
