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FACS
(Fluorescence Activated Cell Sorter) facility was established in 1997
with support from DBT, Govt. of India. There are five flow
cytometers for
advanced scientific research. BD FACScan |
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FACS Committee members
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Dr. William R. Surin, PhD Scientific
Officer Dept.
of Microbiology & Cell Biology Indian
Institute of Science Bangalore-560012 Phone:
+91-80-22933451 Fax:
+91-80-23602697,
Email:
wrsurin@mcbl.iisc.ernet.in |
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FACS Facility members:
Dr.
Omana Joy, Project Associate Ms. Puja Pai, Project Assistant Ms. Kavya A, Project Assistant |
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Flow Cytometers: Analyzers |
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BD FACScan: Single Laser with 3-color parameters. Mac operating system with CellQuestPro Software for acquisition & analysis. Laser Specifications:15mW 488nm, air-cooled argon-ion laser Parameters: Forward Scatter, Side Scatter, FL1, FL2 and FL3
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BD
FACSCaliburTM:It
is dual laser with 4-color parameters. Laser Specifications:15mW 488nm, air-cooled argon-ion laser; Red diode laser, 635nm Fluorescence Detectors and Filters: High performance, high dynamic range photo- multipliers with bandpass filters : 530nm (FITC), 585nm (PE/PI), 661nm (APC) and >650nm (PerCP) with base unit, >670nm (PerCP) with FL4 option Sample Flow rate: Three selectable flow rates of 60mL/min, 35mL/min and 12mL/min. Particle velocity in flow cell is approximately 6 meters/sec Sample Concentration: Single-cell suspension of 105 to 2 x 107 particles/mL recommended range. Kindly use 12 x 75-mm polystyrene tubes (Falcon) (Part No. 54-10001-00) for samples for acquisition. |
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BD FACSCantoTMII: It is configured with 3 lasers and 8 fluorescence parameters. Windows XP operating system with FACS DivaTM Software for acquisition & analysis Laser Specifications: Air-cooled, 20 mW solid state 488 nm, 17 mW HeNe 633 nm laser and 10mW–17mW Point Source Violet solid state laser 405nm Optics configurations: FSC Detector Photodiode with 488/10BP, SSC Detector PMT with 488/10BP Fluorescence Detectors: 8 PMTs in 4+2+2 configuration Four wavelengths detected from the 488-nm laser (standard filters): (1). 515–545 nm FITC (2). 564–606 nm PE (3). 670LP nm PE-Cy™5 or PerCP (4). 750–810 nm PE-Cy™7 Two wavelengths detected from the 633-nm laser (standard filters): (1). 650–670 nm APC (2). 750–810 nm APC-Cy7 Two wavelengths detected from the 405-nm laser (standard filters): (1). 485–535 nm AmCyan (2). 425–475 nm Pacific Blue™ |
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Features: Sample Injection: Direct into flow cell Max Particle Size: 50 μm Flow Rate, min: 10 μL/min Flow Rate, max: 120 μL/min Sample Acquisition Rate: 10,000 events/sec, 10 channels with 8 compensated fluorescent parameters Nominal Minimum Sample Volume: 30 μL |
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Flow Cytometers: Cell Sorters |
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BD FACSAriaTMII: It is having three lasers with 9-color parameters. Windows XP operating system with FACS DivaTM Software for acquisition, analysis and sorting. Laser Specifications: 488-nm Coherent® SapphireTM solid state (13mW–20mW), 633-nm JDS UniphaseTM HeNe air-cooled(10mW–20mW), 375-nm near UV, air launched laser (7–10 mW). All lasers are Class IIIb. Fluorescence Detectors: 9 PMTs in 5+2+2 configuration Five wavelengths detected from the 488-nm laser (standard filters): (1). 515–545 nm FITC, (2). 564–606 nm or 563–589 nm PE, (3). 600–620 nm PE-Texas Red® (4). 675–715 nm or 665–685 nm PE-Cy™5 or PerCP or PI, (5). 750–810 nm PE-Cy™7 Two wavelengths detected from the 633-nm laser (standard filters): (1). 650–670 nm APC (2). 750–810 nm APC-Cy7 Two wavelengths detected from the 375-nm laser (standard filters): (1). 430–470 nm Hoechst blue emission (2). 675 nm Long pass Hoechst red emission Features: Sheath pressure is adjustable from 5 to 75 psi. Filters and mirrors are user changeable. Maximum acquisition rate (events per second) with 12 compensation pairs and 8 parameters: 70,000 events/sec. Purity and Yield At 70 psi and 90 kHz with an average threshold rate of 25,000 events per second, a four-way sort achieved a purity of >98% and a yield >80% of Poisson’s expected yield. Higher threshold rates of up to 70,000 events per second can be achieved without affecting purity; however, yield will decrease based on Poisson statistics. Viability All sorts resulted in cells that proved viable and proliferated for several days post-sort. Sort Collection Devices Two-way sorting: microtubes, 12 x 75 mm, and 15 mL Four-way sorting: microtubes and 12 x 75 mm Automatic Cell Deposition Unit (ACDU) for slide and plate sorting: 6, 24, 48, 96, and 384-well plates Contact person: Dr. Omana Joy |
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Features: Sample Injection Chamber; Sample Input Sizes Microtubes, 12 x 75 mm, and 15 mL Polystyrene or polypropylene tubes can be used. Sample Flow Rates Adjustable sample pressures from 1 to 11 Sample Input Agitation Adjustable through the software to keep sample constantly suspended Temperature Control Sample input, software-adjustable: 4, 20, 37, and 42°C Sample output for sort collection: water refrigerator or heater option is available to provide cooling or heating for sort collection into tube holders, multi-well plates, and slides BD FACS Accudrop • Red diode laser provided for manual or fully automated drop-delay determination • Automated drop break-off monitoring • Automated clog detection system using Sweet Spot technology |
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MoFLo Legacy Cell Sorter & Analyser: It is dual laser lasers (488nm and 350nm) with 5-color parameters. Windows XP operating system with SummitTM Software for acquisition, analysis and sorting. Laser Specifications: 488nm iCyte 30-200mW air cooled solid state laser and 350nm JDSU UV air cooled laser (2 color) Fluorescence Detectors: 5 PMTs in 3+2 configuration Three wavelengths detected from the 488-nm laser: (FL1). 510–550 nm (FL2). 565–605 nm (FL3). 603-623 nm Two wavelengths detected from the 350-nm laser: (FL6). 418–483 nm (FL7). 615–645 nm For details contact: Dr. William R Surin |
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Contact Us: @ Flow
Cytometry Facility Ground
Floor, GCCL-01, Biological Sciences Building, Indian Institute of Science Bangalore-560012, Tel: +91-80-22933451 |
Some useful links: International Society for Advancement of Cytometry
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by wrsurin@mcbl.iisc.ernet.in, Last updated on 26th March, 2012 |
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