Instruments and Applications

Instruments and Applications

The Flow Cytometry Core currently offers five instruments. Two sorters and three analyzers. Click the tab below to view the instrument profile.

Astrios

Astrios

 

Description: The Beckman Coulter Moflow Astrios cell sorter is a digital sorter enabling high-speed sorting and multicolor analysis. Our Astrios has 4 lasers that can detect up to 14 simultaneous colors (see the below chart). We have the capability to sort up to 6 different cell populations simultaneously from a heterogeneous sample. The Astrios allows for sorting onto slides and into plates with up to 1500 wells. Other options include the temperature control unit and the aerosol management unit.

 

Common Applications: The Astrios is particularly well suited for sorting single cells in to 384 well plates that can be used for downstream genomic/metabolomic anslysis as well as clonal expansion. The array of lasers and corresponding detectors enaples sorts from single color GFP to complex 14 color pannels to detect rare cells such as stem cells, cancer stem cells, and other immune cells. Our current hardware does support cell cycle analysis and sorting of cells in G1/G0 and G2 stages along with 6 -way sorting.

 

Scheduling: Review the Flow Cytometry Calendar in iLabs for available times.

 

FACSAria Configuration:

 

Laser
Detector
Band Pass Filter
Colors for Each Channel

405 nm

solid-state laser,

55 mW

FL1
448/59
Pacific Blue/Dye Cycle Violet/v450/DAPI
FL6
546/20

Alexa 430/

v500/Cascade Yellow

405/30
SSC (Side Scatter)

488 nm

Sapphire

solid-state laser, 165mW

FL10
513/26
FITC/AF488/Acridine Orange (DNA)/GFP/CFSE/CDF
FL11
576/21
PE/PI/Cy3
FL12
620/29

PI/PE-TXRed/7-AAD/Ethidium Bromide

FL7
664/22

PE-Cy5/PerCP/Acridine Orange (RNA)/7-AAD

FL9
710/45

PE-Alexa700/PE-Cy5.5/PerCP-Cy5.5

FL8
795/70
PE-Cy7
488/30
SSC (Side Scatter)

561 nm

Yellow Laser

200 mW

FL18
579/16
PE/mTomoto/AL555/RFP/DsRED
FL13
614/20

PE-TxRed/mCherry

FL14
692/75

PE-Cy5/PE-Cy5.5/7-AAD/mPlum

561/30

SSC (Side Scatter)

640 nm

solid-state laser

105 mW

FL23
671/30

APC/AF647/Cy5/DRAQ5

F24
722/40
APC-AF700/APC-Cy5.5/PerCP-Cy5.5
FL19
795/70
APC-AF750/APC-Cy7
640/30
SSC (Side Scatter)

 

FACSAria

 

Description: The BD FACSAria cell sorter is a digital sorter enabling high-speed sorting and multicolor analysis. Our FACSAria has 5 lasers that can detect up to 13 simultaneous colors (see the below chart). We have the capability to sort up to four different cell populations simultaneously from a heterogeneous sample. The ACDU unit allows for sorting into 6, 12, 24, and 96 well plates. Other Aria options include the temperature control unit and the aerosol management unit. Users that have taken the BD Bioscience operators course may use this instrument at the self-user rate.

 

Common Applications: Sorting transgenic cell lines (i.e. GFP cell lines), sorting cell subsets (i.e. regulatory T Cells, macrophages) and multicolor analysis (>4 colors). Our current hardware does support cell cycle analysis and sorting of cells in G1/G0 and G2 stages.

 

Scheduling: Review the Flow Cytometry Calendar in iLabs for available times.

 

FACSAria Configuration:

 

Laser
Detector
Long Pass Filter
Band Pass Filter
Colors for Each Channel

488 nm

Coherent Sapphire solid-state laser,

22 mW

A
655LP
695/40
PerCP/PerCP-Cy5.5/PI/7-AAD
B
502LP
530/30

FITC/CFSE/ALEXA 488/

GFP/Sytox Green/YFP

C
-
488/10
SSC (Side Scatter)

561 nm

Coherent Sapphire

solid-state laser, 50mW

A
735LP
780/60
PE-Cy7
B
630LP
670/14
PE-Cy5/PI/mCherry
C
600LP
610/20

PE-TexasRed(Tandem)/

PI/mCherry/Alexa 568

D
-
582/15
PE/RFP/Alexa 555

633 nm

DS Uniphase Helium Neon laser, 20 mW

A
735LP
780/60
APC-Cy7/APC-H7
B
-
660/20

APC/Alexa 647/

eFluor 660

405 nm**

Coherent VioFlame solid-state laser, 25 mW

A
735LP
780/60

BV785

B
635LP
670/14
BV650
C
595LP
610/20
BV605
D
502LP
525/50
AF430/BV510/V500
E
-
450/50
Pacific Blue/Dye Cycle Violet/v450/DAPI

375nm **

Spectra-Physics, 10mW

Near UV

A
735LP
780/60

BUV805

B
635LP
670/14
BUV661/Hoechst Red (33342)
C
595LP
610/20
BUV563
D
502LP
525/50
BUV496/Zombie UV
E
-
450/50
BUV/395 Hoechst Blue (33342)

 

**NOTE - The 405nm and the 375 nm lasers share the same detectors and electronics. Thus, only ONE of these can be in operation at any given moment.

LSR-II

LSR-II

 

Description: The BD LSR-II is a high-speed, multi-color, digital analyzer. Our LSR-II has 4 lasers that can detect up to 11 different colors (see the below chart). The LSR-II can acquire up to 20,000 events/second. The FACSDiva software allows for automatic compensation setup, and allows for compensation after data acquisition. This minimizes the amount of sample needed to obtain correct compensation before data acquisition. This is an easy-to-operate instrument and is made available to self-users 24/7 as free time permits

 

Common Applications: We can perform 11-color flow cytometry analysis. Our LSRII also has the capabilites to detect very small particles using the Forward Scatter PMT. Using more colors in a given sample can decrease the number of different panels or tubes. This ultimately decreases the amount of total sample needed to collect meaningful data. Also, more colors more efficiently identifies sub-populations from a heterogeneous sample.  Because we can compensate our samples after data acquisition, samples with more difficult compensations (i.e FITC and PI) should be run on the LSR-II. The LSR-II does not have the hardware to sort cells.

 

Scheduling: Review the Flow Cytometry Calendar in iLabs for available times.

 

LSR-II Configuration:

 

Laser
Detector
Long Pass Filter
Band Pass Filter
Colors for Each Channel

405 nm

100 mW

A
630LP
660/20
Brilliant Violet 650
B
505LP
525/50
BV510/V500
C
-
450/50
Pacific Blue/Dye Cycle Violet/v450/BV421

488 nm

Coherent Sapphire solid-state laser, 100 mW

A
685LP
695/40
PerCP/PerCP-Cy5.5/PI/7-AAD
B
505LP
525/50

FITC/CFSE/Alexa 488/

GFP/Sytox Green/YFP

(SSC)
488LP
  SSC (Side Scatter)

561nm

Coherent Sapphire

solid-state laser, 50mW

 

A
750LP
780/60/26
PE-Cy7
B
630LP
660/20

PE-Cy5

C
595LP
610/20
PE-Texas Red
D
-
585/15
PE

633 nm

50 mW

A
750LP
780/60
APC-Cy7/APC-H7
B
-
670/30
APC/Alexa 647/eFluor 660

Dual Forward Scatter

Diode
-
-
Default Scatter
PMT
-
-
For Small Particles

Celesta

LSR-II

 

Description: The BD FACSCelesta is a high-speed, multi-color, digital analyzer. Our Celesta has 3 lasers that can detect up to 12 different colors (see the below chart). The LSR-II can acquire up to 20,000 events/second. The FACSDiva software allows for automatic compensation setup, and allows for compensation after data acquisition. This minimizes the amount of sample needed to obtain correct compensation before data acquisition. This is an easy-to-operate instrument and is made available to self-users 24/7 as free time permits

 

Common Applications: We can perform 12-color flow cytometry analysis. Using more colors in a given sample can decrease the number of different panels or tubes. This ultimately decreases the amount of total sample needed to collect meaningful data. Also, more colors more efficiently identifies sub-populations from a heterogeneous sample.  Despite the inclusion of FACS in the name of the instrument the FACSCelsta does not have the hardware to sort cells.

 

Scheduling: Review the Flow Cytometry Calendar in iLabs for available times.

 

FACSCelesta Configuration:

 

Laser
Detector
Long Pass Filter
Band Pass Filter
Colors for Each Channel

Violet Laser

405 nm

50 mW

A
750LP
780/60
Brilliant Violet 786
B
655LP
670/30
Brilliant Violet 650
C
595LP
610/20
Brilliant Violet 605
D
505LP
525/50
Brilliant Violet510/V500
E
-
450/40
Pacific Blue/Dye Cycle Violet/v450/BV421

Blue Laser

488 nm

20 mW

A
750LP
780/60
PE-Cy7
B
670LP
695/40
PerCP/PerCP-Cy5.5/PI/7-AAD
C
550LP
575/20

PE

D
505LP
530/30

FITC/CFSE/Alexa 488/

GFP/Sytox Green/YFP

(SSC)
-
488/10
SSC (Side Scatter)

Red

640 nm

40 mW

A
750LP
780/60
APC-Cy7/APC-H7
B
750LP
730/45
APC-R700/Alexa Fluor 700
C
-
670/30
APC/Alexa 647/eFluor 660

Forward Scatter

Diode
-
-
Default Scatter

FACSCalibur

 

Description: The BD FACSCalibur is an analogue analyzer equipped with 2 lasers permitting detection of 4 colors (see chart below). This is an easy-to-operate instrument and is made available to self-users 24/7 as free time permits.

 

Common Applications: The best application for the FACSCalibur is cell cycle. All other applications should be run on either the LSRII or the FACSCelesta. Our FACSCalibur does not have the hardware to sort cells.

 

Scheduling: Review the Flow Cytometry Calendar in iLabs for available times.

 

FACSCalibur Configuration:

 

Laser
Detector
Band Pass Filter
Colors for Each Channel
488 Argon (15mw)
FL1
530/30
FITC/CFSE/Alexa 488/GFP/Sytox Green/YFP
FL2
585/42
PE/PI
FL3
670LP (Long Pass)
PerCP/PerCP-Cy5.5/PI/7-AAD/Cy5
635 Red Diode (6mW)
FL4
661/16
APC/Alexa 647/eFluor 660