Welcome!
The Department of Microbiology & Immunology Flow Cytometry Core assists researchers in flow cytometry based studies. Cell samples can be quickly analyzed based on phenotypic markers and functional assays. Characterization of distinct cell populations based on these techniques has become increasingly important in biomedical research. Flow cytometry is especially useful for physically separating specific sub-populations defined by specific parameters using a fluorescence activated cell sorter (FACS). The microscope core provides researchers the ability to obtain high quality imaging data using state-of-the-art microscopy instruments.
Services include:
The shared resource operates three flow cytometers. The LSRFortessa and LSR II are designed for multi-fluorescence analysis while the custom configured FACS Aria II SORP high-speed cell sorter is capable of rapid sorting of cells, bacteria, yeast, and other small particles based on multiple parameter characteristics into highly pure populations. Our two flow cytometers and cell soter have five lasers capable of simultaneous 18-color detection (i.e. 488nm - 2 PMTs, 405nm - 7 PMTs, 355nm - 3 PMTs, 640nm - 3 PMTs, and 561nm - 3 PMTs) of apoptosis, DNA/cell cycle analysis, detection of novel living color fluorescent proteins (DsRed, mCherry, tdTomato, mOrange, mPlum, et al.), immunophenotyping and calcium influx experiments.
M&I Flow Core Instrument Configurations |
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2017 Upgrade |
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All instruments (LSR II, LSR Fortessa, Aria II) have the same configuration |
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Updated LSR II, Fortessa, and Aria II Configurations |
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| **Optional Configuration** (Filter/Configuration Change Required)# | ||||||||
| Laser | Channel Name | PMT Position | Dichroic | Bandpass | Similar Parameter (no filter change required)# |
Parameter/Stain | Dichroic | Bandpass |
| UV 355 nm |
BUV395 | C | - | 379/28 | DAPI, Indo-1 (violet) | - | 450/50 | |
| BUV496 | B | 450 LP | 515/30 | Fixable Live/Dead Aqua | Indo-1 (blue), Qdot 525 | 505 LP | 530/30 | |
| BUV737 | A | 690 LP | 735/35 | |||||
| VIOLET 405 nm |
BV421 | G | - | 450/50 | Pacific Blue, eFluor450, violetFluor450, V450, Alexa405 | |||
| BV510 | F | 505 LP | 525/50 | AmCyan, Pacific Orange$, V500, Alexa430 | ||||
| BV570 | E | 550 LP | 575/25 | PacificOrange$ *this channel can be difficult to compensate* |
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| BV605 | D | 595 LP | 610/20 | Qdot 605 | ||||
| BV650 | C | 635 LP | 670/30 | Qdot 655 | ||||
| BV711 | B | 690 LP | 710/50 | Qdot 705 | ||||
| BV785 | A | 750 LP | 780/60 | Qdot 800 | ||||
| BLUE 488 nm |
SSC | C | - | 488/10 | ||||
| FITC | B | 505 LP | 525/50 | GFP, Alexa488, BrilliantBlue515 | ||||
| PerCP-Cy5.5 | A | 685 LP | 710/50 | PE-Cy5.5, PerCP-eFluor710 | ||||
|
YELLOW/ GREEN |
PE | C | - | 582/15 | DsRED, dTomato | |||
| PE-TexasRed | B | 600 LP | 610/20 | PE-eFluor610, PE-CF594, mCherry, 7-AAD, Alexa568 | ||||
| PE-Cy7 | A | 750 LP | 780/40 | |||||
| RED 640 nm |
APC | C | - | 670/30 | Alexa647 | |||
| AF700 | B | 685 LP | 730/45 | redFluor 710 | ||||
| APC-Cy7 | A | 750 LP | 780/60 | APC-eFluor780, APC-H7 | ||||
| #compensations must be made with the same conjugate/stain used within each experiment |
| **For DAPI/Indo-1 please see Flow Core Manager about filter/configuration change** |
| $due to the broad emmision spectrum of Pacific Orange, both channels will detect this fluor |
The flow cytometric separation of cells from unfixed animal tissue is possible on the BD FACSAria cell sorter, which is housed in a Baker BioProtect IV biohazard cabinet. For unfixed human specimens, please contact Allyson Capili. Before the initial sort is performed, each user is required to meet with the core manager to discuss your cell type, origin of sample, and special requirements.
In additional to these systems, the facility also has a state-of-the-art Zeiss LSM710 ConfoCorr confocal imaging microscope system, a Zeiss AxioVert microscope, and a Typhoon Trio imaging system.
Ryan Wantroba
Flow Core Manager
Phone: 203-233-8665
Email: rmw2178@cumc.columbia.edu
Nicholas Arpaia, Ph.D.
Director
Phone: 212-305-1379
Email: na2697@cumc.columbia.edu
James Lapin
Dept. Administrator
Phone: 212-305-4046
Email: jl2787@cumc.columbia.edu
Carol Duigou
Accounts Manager
Phone: 212-305-3647
Email: ced4@cumc.columbia.edu
| Hours | Location |
|
Staffed |
Department of Microbiology & Immunology |
| Name | Role | Phone | Location | |
|---|---|---|---|---|
| Microbiology & Immunology Shared Resources |
Managers
|
212-342-4089
|
microimmuno-facs@cumc.columbia.edu
|
HHSC 1211-A
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