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Analytical Cytometry

Analytical Cytometry Core (ACC)
City of Hope’s Analytical Cytometry Core (ACC) Facility provides researchers with high-quality flow cytometry instrumentation, as well as expertise in analyzing and/or sorting sample populations of interest via interpretation of their physical, fluorescent and/or light-scattering properties. The facilities and their services are available to both City of Hope and non-City of Hope researchers.
 
The ACC Facility has two components: the Flow Cytometry Facility (FCF) and the Molecular Imaging Facility (MIF).
 
As an added convenience, the ACC supports several off-line workstations throughout the campus where investigators can analyze their ACC-derived data. FCF software includes Flowjo (only on some workstations), Modfit (only on some workstations)and Summit. MIF software includes ImageQuant, PDQuest (Bio-Rad) or Odyssey.
 
Flow Cytometry Facility
The Flow Cytometry Facility has three Cell sorters (MoFlo, Aria III, and Aria SORP) and 5 analyzers (CyAn, Gallios, Fortessa, C6, and Guava). All the analyzers are available for unassisted end user runs.
 
Research reported in this publication included work performed in the Analytical Cytometry Core supported by the National Cancer Institute of the National Institutes of Health under award number P30CA33572. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Analytical Cytometry Equipment

We offer the following equipment in the Analytical Cytometry Core.
 
Flow Cytometry Facility (FCF):
  • MoFlo™ MLS
  • Aria III
  • Aria SORP
  • CyAn™ ADP 9 Color
  • Gallios
  • Fortessa
  • C6
  • Robo Sep Nagnetic Bead Separator
 
 

Aria SORP

The Aria SORP (Special Order Research Product) is a six laser system (355nm, 405nm, 457nm, 488nm, 561nm & 640nm) with a maximum of 17 simultaneous fluorescent detectors.  (See tech specs) But the instrument is configured to have 19 fluorescent detectors to choose from. In addition to a standard forward scatter photodiode detector, the Aria SORP is equipped with a Photomultiplier tube (PMT) for forward scatter.  This additional parameter allows for detection of particles as small as 0.4 microns.  Samples can be loaded onto the sorter in a variety of tube sizes (microfuge tubes, 12X75mm or 15ml conical centrifuge tubes).  The possible nozzle tip sizes available for sorting are 70 micron, 85 micron, 100 micron, and 130 micron.  Bulk sorting of 1 to 4 populations per sample is possible.  Collection tubes for 1 or 2 way sorting are 15ml, 12X75mm, or 1.5ml microfuge tubes.  With 3 or 4 way sorting, either 12X75mm or 1.5ml microfuge collection tubes are utilized.  Also, an Automatic Cell Deposition Unit (ACDU) is available for sorting onto slides and plates (6, 24, 48, 96 or 384-well plates).  With the ACDU single cell or any user specified number of cells can be collected.  The Aria SORP is equipped with temperature control for both the sort sample and the sort collection tubes.  This sorter is contained in a Biosafety Cabinet.
 
Quality Control

Cytometer Setup and Tracking (CS&T) beads are purchased from BD Bioscience and run on a daily basis to monitor system performance.   Also Accudrop beads (BD Bioscience) are utilized to setup the sorting parameters of the system prior to cell sorting for a given day.
 
Supplies Needed

Investigators must supply their own reagents.  All samples are filtered by core staff prior to placement on the cell sorter.  We utilize 60micron nylon mesh that has been autoclaved for sterility.  It is recommended that the user provides collection tubes with the appropriate media for their specific cell type.  For plate sorting, the user must provide the appropriate plate type and collection media.  It is also recommended that the user bring extra collection media.

C6

C6
Document is not available. A template could not be found for CoH Media 1365762911429.The C6 is a 2 laser (488nm and 640nm) system.  From the 488nm laser up to 3 fluorescent detectors are available and only one detector for the 640nm laser.  This cytometer is available for trained users to operate.
 
Quality Control
Spectralign  beads are purchased from Spherotech and run on a monthly basis to monitor system performance.  
     
Supplies Needed
Investigators must supply their own reagents.  Users that are trained to self operate the instrument will need to provide their own tubes.  This instrument requires BD-Falcon polysytrene 12X75mm tubes for operation.

CyAn

The CyAn™ ADP 9 Color has three excitation sources: 405nm, 488nm & 635nm. This instrument has 11 parameter capabilities (two scatter and nine fluorescent detectors). All parameters can be collected in linear peak height, linear area, and/or log scales. It is capable of acquisition rates up to 50,000 events per second. This cytometer is available for trained users to operate, or user samples may be analyzed by the core at an additional cost.
 
 
CyAn™ (Flow Cell) CyAn™ ADP
(Advanced Digital Processing)
Quality Control
Spherotech Ultra-Rainbow Spectralign beads are run once a week to ensure proper laser alignment for stability and reproducibility.
 
Supplies
Investigators must supply their own reagents.  Users that are trained to self operate the instrument will need to provide their own tubes.  This instrument requires polypropylene or  polysytrene 12X75mm tubes for operation.
 

Fortessa

The Fortessa is a 5 laser analytical cytometer.  This cytometer is a special order research product (SORP) and is equipped with 355nm (UV), 405nm, 488nm, 561nm and 640nm lasers. (See tech specs)  Laser power can be adjusted with software.  Up to 17 simultaneous fluorescent parameters can be detected.  Link for the standard configuration is attached  Along with standard single tube loading, a device for the introduction of samples from a 96 or 384 well format is available (BD-high throughput sampler-HTS).  This cytometer is available for trained users to operate, or user samples can be analyzed by the core staff at an additional cost.
 
Quality Control
Cytometer Setup and Tracking (CS&T) beads are purchased from BD Bioscience and run on a daily basis to monitor system performance.     
 
Supplies Needed
Investigators must supply their own reagents.  Users that are trained to self operate the instrument will need to provide their own tubes.  This instrument requires BD-Falcon (Ref# 352008) polysytrene 12X75mm tubes for operation.

Gallios

The Gallios is an analytical cytometer with 3 lasers and 10 fluorescent detectors (405nm-2 detectors, 488nm-5 detectors and 638nm-3 detectors).  Attach the parameter list.  There are 3 settings for forward scatter detection to increase the resolution.  A 32 tube carousel provides for setup and walk away sample acquisition.  All parameters can be collected in linear and/or logarithmic scales as height, area and/or width measurements.  This cytometer is available for trained users to operate, or user samples can be analyzed by the core staff at an additional cost.
 
Quality Control
Flow Check Pro standard beads are purchased from Beckman Coulter and run on a weekly basis to monitor system performance.  
 
Supplies Needed
Investigators must supply their own reagents.  Users that are trained to self operate the instrument will need to provide their own tubes.  This instrument requires 12X75mm tubes for operation.

MoFlo MLS

The MoFlo™ MLS has a three-laser optical bench configuration.  The three lasers (one argon gas and 2 krypton gas) provide fluorescent excitations from 351-752.5 nm.  This instrument has 10 separate photomultiplier tubes (PMTs) plus a forward scatter detector.  Peak height, integrated and logarithmic data can be collected from each of the PMTs.  There are optical filters for the collection of up to 9 simultaneous fluorescent signals.  The facility normally sorts with a 100 micron nozzle at 30 PSI.  Up to four sort decisions can be performed simultaneously.  Along with bulk sorting, cell deposition into wells as single cells or multiple cells is also a possibility.  The instrument can accommodate plates that contain from 6 to 384 wells.
 
MoFlo: 4-way sorting streams MoFlo: Cytomation sorting unit
MoFlo MLS (Multi-Laser System)  
 
Quality Control
UltraRainbow Fluorescent Particles are purchased from Sphereotech and run on a daily basis to monitor system performance.   These same beads are utilized to setup the sorting parameters of the system prior to cell sorting for a given day.
 
Supplies Needed
Investigators must supply their own reagents.  All samples are filtered by core staff prior to placement on the cell sorter.  We utilize 60micron nylon mesh that has been autoclaved for sterility.  It is recommended that the user provides collection tubes with the appropriate media for their specific cell type.  For plate sorting, the user must provide the appropriate plate type and collection media.  It is also recommended that the user bring extra collection media.
 

Odyssey® Infrared Imager

The Odyssey® Infrared Imager scanner can be used to analyze two-color western blots, two-color in-gel westerns, two-color northern blots, and any other primary antibody tagged samples, which can be stained with near-infrared secondary antibodies (IRDye 700 and IRDye 800) or Alexa Fluor 680.  It can also visualize Coomasie stained protein gels, which are fluorescent in the infrared.  Along with scanning gels and membranes, it can also scan microplate cell-based assays.  Various resolutions can be used for scanning, which range from 21u to 337u.
 
Quality Control
Users are generally responsible for running their own controls.
 
Supplies
Cleaning solution and Kimwipes® are provided for cleaning the platen on the scanner.  All other reagents, solutions, buffers, and supplies must be provided by the user.

RoboSep

The RoboSep is a magnetic bead based separation unit which will allow for either positive or negative selection of up to 4 distinct samples without the use of columns.  This unit is located inside a biosafety cabinet for sterility of the samples.  All supplies must be purchased by the investigator.
 
 

Typhoon™ 9410

The Typhoon™ 9410 equipment is available to scan phosphor screens, which have been exposed to radioactive sources on gels/blots/dishes/trays/glass slides.  The Analytical Cytometry Core has a large format (35x40 cm) phosphor screen and several small (20x25 cm) format screens available for proof of methodology.  Screen erasers are located at the core workstations or nearby.  This allows linear quantification of data (over five orders of magnitude) and produces digital images, which are directly manipulatable for analysis and publication.  Exposure times are usually 20-25% of those required on film.  Radioisotope sources have included P32, S35, C14, and I125. There are also special screens available from Amersham Biosciences for tritium detection.
 
The Typhoon™ 9410 also has the capability to detect fluorescent labels on gels, blots, dishes, trays, and glass slides. This instrument has four excitation sources: 457nm, 488nm, 532nm, and 633nm that allows for use of a wide range of fluorescent dyes.  There are also selectable emission filters (520BP40, 555BP20, 580BP30, 610BP30, 670BP30, 526SP, and 560LP), which allow for collection of specific and multiple signals.  This methodology allows for linear quantification of the data (over three to four decades of magnitude) and produces digital images, which are directly manipulatable for analysis and publication.  The matrix of interest can be scanned at various resolutions depending on experimental design.  For radioactivity, the range in resolution is from 25u to 1000u and for fluorescence, the range is from 10u-1000u.  At 10u the scanner is capable of handling microarray slides.
 
Quality Control
Users are generally responsible for running their own controls.
 
Supplies
Cleaning solution and Kimwipes® are provided for cleaning the platen on the scanner.  All other reagents, solutions, buffers, and supplies must be provided by the user.  Several phosphor screens are provided by the core for radioisotope scans, but users are encouraged to buy their own screens.

Using the Facility

Scheduling Appointments
 
Flow Cytometry Facility
 
To schedule appointments it is best email Lucy Brown (lbrown@coh.org).
 
Turn-around Time
 
Simple analysis schemes are accomplished as samples are run.If more detailed analysis is needed, it is usually done within a 24-hour period.
 
 
 

Abstract for Grants

The Analytical Cytometry Core (ACC) is comprised of two facilities: the Flow Cytometry Facility and the Molecular Imaging Facility. Both facilities are available for every investigator of the Beckman Research Institute of City of Hope.
 
The Flow Cytometry facility contains 3 cell sorters and 4 analytical cytometers.
 
The cell sorters include:
  • MoFlo legacy (Beckman Coulter) with 3lasers (405nm, 488nm & 640nm) and up to 9 parameters
  • Aria III (Becton Dickinson) with 4 lasers (405nm, 488nm, 561nm & 633nm) and up to 15 fluorescent parameters
  • Aria II SORP (Becton Dickinson) with 6 lasers (355nm, 405nm, 457nm, 488nm, 561nm & 640nm) and up to 18 simultaneous fluorescent parameters. This sorter is contained in a biosafety cabinet

 

The Analytical cytometers include:
  • Gallios (Beckman Coulter) with 10 fluorescent detectors and 3 lasers (488nm-5detectors, 405nm-2detectors, & 638nm-3detectors). It also has a 32 tube carousel for walk away data acquisition.
  • CyAn ADP (Beckman Coulter) with 9 fluorescent detectors and 3 lasers (488nm-5detectors, 405nm-2detectors, & 640nm-2detectors). We also have the ability to attach a Hypercyte for high throughput screening from 96/384 well platform.
  • Fortessa SORP (Becton Dickinson) with 5 lasers (355nm, 405nm, 488nm, 561nm, 640nm) and up to 17 simultaneous fluorescent parameters. HTS module for 96 well sample delivery allowing for automation is also available.
  • C6 (Becton Dickinson) with 4 fluorescent detectors and 2 lasers (488nm-3 detectors, 640nm-1 detector).

 

ACC also provides training for user operation of the analytical instruments, basic flow theory, software, and data analysis for result generation and presentation. Other functions of ACC include application assistance, experimental design, and site license management for Flowjo and FCSexpress.
 
 
 

Pricing

Prices and availability vary. Please contact us for current information.
 
If you are a City of Hope employee, please visit this core's intranet site for pricing.
 

Contact Us

Jeremy Stark, Ph.D.
Director
626-256-HOPE (4673), ext. 63346
jstark@coh.org
 
Lucy Brown, M.S.
Staff Scientist
626-256-HOPE (4673), ext. 67172
lbrown@coh.org
 
Alexander Spalla, B.S.
Research Associate II
626-256-HOPE (4673), ext. 65832
aspalla@coh.org
 
Ni Feng
Research Associate II
626-256-HOPE (4673), ext. 65832
nifeng@coh.org
 
Lippman-Graff Building:
Room 101 - MoFlo Cell Sorter
Room 102 - Tissue Culture for Flow Cytometry Facility
Room 103 - Supply Room and Preparation Room for MIF
Room 114 - Molecular Imaging Facility Scanners
Room 221 - Managers Office
Room 233 - Staff Office
Room 227 - CyAn and Gallios Analyzer
Room 228A - Fortessa Analyzer
Room 228B - Robo Sep Magnetic Bead Separator
Room 229 - AriaIII, and Aria SORP Cell Sorter
 
Gonda Building:
Room 1006B - BD Fortessa Cytometer
Room 3113 - BD C6 Analyzer
 
Beckman Research Center:
Room 3300– BD C6 Analyzer
 
 
Research Shared Services

City of Hope embodies the spirit of scientific collaboration by sharing services and core facilities with colleagues here and around the world.
 

Recognized nationwide for its innovative biomedical research, City of Hope's Beckman Research Institute is home to some of the most tenacious and creative minds in science.
City of Hope is one of only 41 Comprehensive Cancer Centers in the country, the highest designation awarded by the National Cancer Institute to institutions that lead the way in cancer research, treatment, prevention and professional education.
Learn more about City of Hope's institutional distinctions, breakthrough innovations and collaborations.
Support Our Research
By giving to City of Hope, you support breakthrough discoveries in laboratory research that translate into lifesaving treatments for patients with cancer and other serious diseases.
 
 
 
 
Media Inquiries/Social Media

For media inquiries contact:

Dominique Grignetti
800-888-5323
dgrignetti@coh.org

 

For sponsorships inquiries please contact:

Stefanie Sprester
213-241-7160
ssprester@coh.org

Christine Nassr
213-241-7112
cnassr@coh.org

 
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