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

ARIA II configuration diagram

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 

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

SSC

FITC / Alexa488 / GFP / YFP / CSFE (530/30)

PE_Blue (575/26)

PerCP-Cy5.5 / PI / 7AAD (695/40)

Violet Laser (405nM) x 3 parameters

BV510 / AmCyan / Aqua Live/Dead (525/50)

BV421 / PacBlue / DAPI/ CellTraceViolet (450/50)

BV605 or 655 / 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 / -H7 / Alexa750 or 780 (780/60)

Green Laser (532nM) x 5 parameters

PE-Gr (585/42)

PE-TxRed / dsRed / mCherry (610/20)

PE-Cy5 / PerCP (670/30)

PE-Cy5.5 / PerCP-CY5.5 (730/45)

PE-Cy7 (780/60)

Numbers in parentheses refer to emission filter wavelengths of each detector

*Contact facility personnel for instructions

LSR14 configuration diagram

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 3 parameters

SSC

FITC / Alexa488 / GFP / YFP / CSFE (525/50)

PerCP-Cy5.5 / PI / 7AAD (710/50)

Violet Laser (405nM) x 8 parameters

BV421 / PacBlue / CellTraceViolet / eFluor (450/50)

BV510 / AmCyan / Aqua Live/Dead (525/50)

BV570 / Qdot 565 (560/40)

Qdot585 (585/42)

BV605 / Qdot 605 / ZombieYellow (605/40)

BV650 / Qdot 655 (660/40)

BV711 / Qdot 705 (705/70)

BV785 / Qdot 800 (780/60)

Red Laser (640nM) x 3 parameters

APC / Alexa647 (660/20)

Alexa700 / APC-Cy5.5 (710/50)

APC-Cy7 / -H7 / Alexa750/790/ APC-eFluor780 (780/60)

Green Laser (532nM) x 5 parameters

PE-Gr (575/25)

PE-TxRed / dsRed / mCherry / PI (610/20)

PE-Cy5 / PerCP / 7AAD (660/20)

PE-Cy5.5 (710/50)

PE-Cy7 (780/40)

Numbers in parentheses refer to emission filter wavelengths of each detector

LSR18 configuration diagram

Contact facility with questions regarding color combinations

 

CyAn ADP, DakoCytomation (purchased 2002) (Hill 480)

● Capable of user-friendly analysis
● Three 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 / GFP / 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.