Flow Cytometry
Overview
The facility has four analytical flow cytometers (BD LSRFortessa and Fortessa X20, Agilent NovoCyte 3000, and a Cytek Aurora full spectrum analyzer) capable of analyzing over 40 colors, all equipped with plate loaders. A BD FACSAria cell sorters with up to 16-color capability is housed in a biosafety enclosures, as are the X20 and Aurora analyzers. The Core also has an imaging flow cytometer (Amnis ImageStreamX MarkII) with up to 10 colors. Cell sorting is performed by expert Core staff with analytical cytometry available either as a service or for use 24/7 by trained investigators. The Core also offers a validated 27-marker antibody panel for spectral profiling of the murine tumor immune microenvironment. FlowJo analysis software is available on workstations in the core and through an institutional site license.
Scientists planning a flow cytometry experiment are encouraged to consult facility staff for assistance with protocols, fluorochrome selection or other aspects of experiment design. Sorter and analyzer optical configurations can be viewed here.
Services
High Speed Cell Sorting
- Sorting into tubes, 96 or 384-well plates for culture or single cell sequencing
- Expertise in calcium flux, cell cycle, side population, and sorting of IPSC and ESC, tumor progenitor and tissue stem cells
- Performed by expert facility personnel
- Simultaneous sorting of up to 4 populations
- BSL2 biosafety enclosure
Analytical Flow Cytometry
- Consultation
- Experiment design
- Hardware and software training
- Do-it-yourself 24hr/day, or assisted by appointment up to 30 colors with plate loader
- Data analysis and interpretation
- Publication quality figures
- Assistance with methods sections
- Pre-publication manuscript review
Validated Antibody Panels
- Murine Tumor Immune Profile (mTIME) Panel
- Core-developed 27-color spectral antibody panel for profiling the murine tumor immune microenvironment
- 27 cell surface markers, titrated reagents, validated staining protocol
- Resolves B cells, T cells (CD4, CD8, activation, Treg, inhibitory receptors), NK, NKT, pDC, cDC1 and cDC2, monocytes, transitional monocytes, macrophages (alternative activation)
- Six spectral drop-in zones for project-specific customization
- Compatible with cells expressing GFP, YFP, TdTomato, mCherry, or BFP
- Available in 10-test increments complete with all necessary buffers.
Equipment & Resources
Cell Sorters
- BD FACSAriaIIu 16-color high-speed cell sorter
- Four solid state lasers (405 nm, 488 nm, 561 nm, 640 nm)
- BSL2 containment
- Sort into tubes, 96 or 384-well plates
Flow Cytometry Analyzers
- Cytek Aurora full spectrum cytometer
- Collects cellular fluorescence across the full visible spectrum
- Allows for highly multiplexed panels with far more colors than a traditional cytometer
- Autofluorescence can be unmixed to improve sensitivity for highly autofluorescent cells
- Acea NovoCyte 3000 13-channel analytical flow cytometer
- 488nm, 405 nm and 640 nm lasers
- Multi format autosampler
- BD LSR Fortessa X20 18-color analytical flow cytometer
- Five solid state lasers (488nm, 637nm, 405nm, 561nm and 355nm)
- BSL2 containment
- HTS plate loader for 96 or 384-well plates
- BD LSRFortessa 14-color analytical flow cytometer
- HTS plate loader for 96 or 384-well plates
- Four solid state lasers (405 nm, 488 nm, 561 nm, 640 nm)
Imaging Flow Cytometers
- Amnis ImageStreamX MarkII imaging flow cytometer
- 12 imaging channels (10 colors and 2 brightfield channels)
- 405 nm, 488 nm and 642 nm laser lines
- 20x, 40x and 60x objectives
- 96-well plate loader
- Extended Depth of Field (EDF) option for spot counting
- Inspire acquisition software
- Ideas analysis software
Computing
- FlowJo site license
- Computer workstations with large monitors and data analysis software
- FlowJo
- ModFit LT
- SpectroFlo
- IDEAS
Price List
For a complete list of services, please call (858) 646-3106 or email us.
| Flow Cytometry Services | Internal Subsidized | Internal | External Non-Profit | External For-Profit | |
|---|---|---|---|---|---|
| FACSAria Sorting, 1st hour | each | $164 | $205 | $221 | $431 |
| FACSAria Sorting, additional hours | hour | $99 | $124 | $134 | $260 |
| ImageStreamX II + Staff Operator | hour | $85 | $107 | $115 | $224 |
| LSRFortessa Independent Use | hour | $35 | $44 | $47 | $92 |
| LSRFortessa + Staff Operator | hour | $96 | $120 | $130 | $252 |
| Aurora + Staff Operator | hour | $108 | $135 | $146 | $284 |
| mTIME Panel | 10 tests | $650 | $813 | $878 | $1710 |
| Data Analysis | hour | $84 | $105 | $113 | $221 |
Contact
The flow cytometry core staff has decades of experience in NCI-designated Cancer Center flow cytometry core labs.
Yoav Altman received his BA degree in Integrative Biology from the University of California at Berkeley. He started working with flow cytometry in 2001 at La Jolla Institute for Allergy and Immunology and joined Sanford Burnham Prebys Medical Discovery Institute in 2002 as Manager of the Flow Cytometry Shared Resource. He was promoted to Director in 2009, and oversees lab operation. Yoav has over 20 years of experience sorting a variety of cell types including IPSCs, hESCs, immune cells, disaggregated tumors and solid tissues such as cardiomyocytes, hepatocytes, intestinal epithelium, brain tissue, and skeletal muscle. His experience includes a variety of single cell assays including side population, cell cycle, apoptosis determination, high parameter immunophenotyping, calcium flux, mitochondrial membrane potential, ROS, fluorescent proteins, and FRET. He is also well versed in optimizing sorting protocols for single cell NGS sequencing using 10x or similar platforms. His current interests include imaging flow cytometry for detection and evaluation of extracellular vesicles and cell-cell interactions. Yoav is available to assist with reagent selection, experiment design, data analysis, protocol development, grant preparation, and writing methods sections for publications
Please call Yoav at (858) 646-3106 or use the button below to send us an email.