ISAC Recognized Shared Resource Laboratory

What we do

The Flow and Immune Analysis Shared Resource (FIASR) provides a comprehensive array of cutting-edge technologies on a variety of platforms including flow and image cytometry, proteomics, metabolomics, and spatial profiling techniques in support of Roswell investigators. The FIASR is recognized by the International Society for the Advancement of Cytometry (ISAC) for operational excellence and adherence to best practices. The Shared Resource is committed to expanding access to unique technologies and the use of advanced instrumentation, while maintaining an active program to educate the scientific community on innovative applications.

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Instruments

Helpful links

Flow cytometry analyzers

The flow cytomety analyzers are available 24/7 to trinaed users by reserving time on the FOM calendar.

Fortessa A,B and C
  • 3L, 16 fluorescent parameters with 2 scatter parameters
  • High Throughput Sampler (96 well plate) option)

Laser

Bandpass Filter

Common Fluors (list is not exhaustive)

405 nm violet laser

V450/50

BV421, BV480, DAPI, Pacific Blue, V450, EF450, SB436, AF405, LD Violet

V525/50

BV510, V500, AmCyan, LD Aqua

V560/20

Qdot565, BV570

V585/15

BV570

V610/20

BV605

V660/20

BV650

V710/50

BV711

V780/60

BV750, BV786, BV785

488 nm blue laser

B530/30

FITC, BB515, GFP, AF488, CFSE

B575/26

PE, mOrange, AF568

B610/20

PECF594, PEAF610, PEDazzle594, PEeFluor610, ECD, PI

B695/40

PerCPCy5.5, BB700, PerCP, PECy5, PECy5.5 7AAD, PEeFluor710

B780/60

PECy5, PEAF750

640 nm red laser

R670/14

APC, AF647

R730/45

AF700, APC-R700, DRAQ5

R780/60

APC-Cy7, APC-AF750, APCH7

Fortessa A,B and C
  • 4L, 18 fluorescent parameters with 2 scatter parameters
  • High Throughput Sampler (96 well plate) option

Laser

Bandpass Filter

Common Fluors (list is not exhaustive)

355 nm UV laser

U379/28

BUV395, Indo-1

U460/14

DAPI, Hoechst 33342, LD Blue, Zombie UV

U740/35

BUV737

405 nm violet laser

V450/50

BV421, BV480, DAPI, Pacific Blue, V450, EF450, SB436, AF405, LD Violet

V525/50

BV510, V500, AmCyan, LD Aqua

V585/15

BV570

V610/20

BV605

V660/20

BV650

V710/50

BV711

V780/60

BV750, BV786, BV785

488 nm blue laser

B530/30

FITC, BB515, GFP, AF488, CFSE

B575/26

PE, mOrange, AF568

B610/20

PECF594, PEAF610, PEDazzle594, PEeFluor610, ECD, PI

B695/40

PerCPCy5.5, BB700, PerCP, 7AAD

B780/60

PECy7, PEAF750

640 nm red laser

R670/14

APC, AF647

R730/45

AF700, APC-R700, DRAQ5

R780/60

APC-Cy7, APC-AF750, APCH7

Fortessa A,B and C
  • 4L, 18 fluorescent parameters with 2 scatter parameters

Laser

Bandpass Filter

Common Fluors (list is not exhaustive)

355 nm UV laser

U379/28

BUV395, Indo-1

U460/14

DAPI, Hoechst 33342, LD Blue, Zombie UV

U740/35

BUV737

405 nm violet laser

V450/50

BV421, BV480, DAPI, Pacific Blue, V450, EF450, SB436, AF405, LD Violet

V525/50

BV510, V500, AmCyan, LD Aqua

V585/15

BV570

V610/20

BV605

V670/30

BV650

V710/50

BV711

V780/60

BV750, BV786, BV785

488 nm blue laser

B530/30

FITC, BB515, GFP, AF488, CFSE

B575/26

PE, mOrange, AF568

B610/20

PECF594, PEAF610, PEDazzle594, PEeFluor610, ECD, PI

B710/50

PerCPCy5.5, BB700, PerCP, PECy5, PECy5.5, 7AAD, PEeFluor710

B780/60

PECy7, PEAF750

640 nm red laser

R670/30

APC, AF647

R710/50

AF700, APC-R700, DRAQ5

R780/60

APC-Cy7, APC-AF750, APCH7

Fluorochrome

Filter available

Calcium Flux

405/20

Side Population

645LP

YFP

546/20

Aurora A, B and C
  • 5L, 64 detectors, 40+ fluorescent parameters with 3 scatter parameters (FSC (488nm), SSC (405nm), SSC-B (488nm)
  • Aurora C has High Throughput Sampler (96 well plate or 40 tube) option

Laser

Channel

Bandpass Filter

Common Fluors (list is not exhaustive)

355 nm UV laser

UV1

NUV373/15

UV2

NUV388/15

BUV395

UV3

NUV428/15

UV4

NUV443/15

UV5

NUV458/15

UV6

NUV473/15

UV7

NUV542/28

BUV496

UV8

NUV542/28

UV9

NUV582/31

BUV563

UV10

NUV613/31

UV11

NUV664/27

BUV661

UV12

NUV692/28

UV13

NUV720/29

BUV737

UV14

NUV750/30

UV15

NUV780/30

UV16

NUV812/34

BUV805

405 nm violet laser

V1

V428/15

BV421

V2

V443/15

AF405, SB436

V3

V458/15

PacBlue, eFluor450

V4

V473/15

BV480

V5

V508/20

eFluor506

V6

V525/17

V7

V542/17

BV510

V8

V581/19

BV570, PacOrange

V9

V598/20

V10

V615/20

BV605, SB600

V11

V664/27

BV650, SB645

V12

V692/28

V13

V720/29

BV711, SB702

V14

V750/30

BV750

V15

V780/30

BV785, BV786

V16

V812/34

488 nm blue laser

B1

B508/20

BB515

B2

B525/17

FITC, AF488

B3

B542/17

AF532

B4

B581/19

B5

B598/20

B6

B615/20

B7

B661/17

B8

B679/18

PerCP

B9

B697/19

BB700, PerCPCy5.5

B10

B717/20

PerCP-eFluor710

B11

B738/21

B12

B760/23

B13

B783/23

B14

B812/34

561 nm yellow/green laser

YG1

YG577/20

PE

YG2

YG598/20

YG3

YG615/20

PEDazzle594, PECF594, PE-AF610, PE-eFluor610

YG4

YG661/17

YG5

YG679/18

PECy5

YG6

YG697/19

YG7

YG720/29

PECy5.5, PE-AF700

YG8

YG750/30

YG9

YG780/30

PECy7

YG10

YG812/34

638 nm red laser

R1

R661/17

APC

R2

R679/18

AF647, eFluor660

R3

R697/19

R4

R717/20

AF700, APC-R700

R5

R738/21

R6

R760/23

R7

R783/23

APCCy7, APCH7, APC-AF750, APCFire750

R8

R812/34

Imaging flow cytometers

ImageStream Mk II
  • 5L, up to 10 fluorescent parameters with two brightfield images
  • 12 channel, 2 camera instrument
  • Side scatter in channel 6 or channel 12 (dedicated 785nm scatter laser)
  • 20x, 40x, 60x magnification
  • Extended Depth of Field (EDF, 16 µ) and autosampler option (96-well plate format)

Camera 1

Bandpass Filter

Common Fluors (list is not exhaustive)

488 nm blue
561 nm yellow/green
592 nm orange

Ch1: 420-480 nm

Brightfield

Ch2: 480-560 nm

FITC, BB515, GFP, AF488, CFSE

Ch3: 560-595 nm

PE, AF568

Ch4: 595-642 nm

PECF594, PEAF610, PEDazzle594, PEeFluor610, ECD, AF594

Ch5: 642-745 nm

PerCPCy5.5, BB700, PerCP, PECy5, PECy5.5, 7AAD, PEeFluor710

Ch6: 745-800 nm

PECy7, PEAF750

Camera 2

Bandpass Filter

405 nm violet
640 nm red

Ch7: 420-505 nm

BV421, BV480, DAPI, Pacific blue, V450, LD Violet

Ch8: 505-570 nm

BV510, V500, AmCyan, LD Aqua

Ch9: 570-595 nm

Brightfield

Ch10: 595-642 nm

BV605

Ch11: 642-745 nm

AF647, APC

Ch12: 745-800 nm

APC-Cy7, APC-AF750, APCH7

ImageStream Mk II
  • 4L, up to 10 fluorescent parameters with two brightfield images
  • 12 channel, 2 camera instrument
  • Side scatter in channel 6 or channel 12 (dedicated 785nm scatter laser)
  • 20x, 40x, 60x magnification

Camera 1

Bandpass Filter

Common Fluors (list is not exhaustive)

488 nm blue
561 nm yellow/green

Ch1: 420-480 nm

Brightfield

Ch2: 480-560 nm

FITC, BB515, GFP, AF488, CFSE

Ch3: 560-595 nm

PE, AF568

Ch4: 595-642 nm

PECF594, PEAF610, PEDazzle594, PEeFluor610, ECD, AF594

Ch5: 642-745 nm

PERCPCy5.5, BB700, PerCP, PECy5, PECy5.5, 7AAD, PEeFluor710

Ch6: 745-800 nm

PECy7, PEAF750

Camera 2

Bandpass Filter

405 nm violet
640 nm red

Ch7: 420-505 nm

BV421, BV480, DAPI, Pacific blue, V450, LD Violet

Ch8: 505-570 nm

BV510, V500, AmCyan, LD Aqua

Ch9: 570-595 nm

Brightfield

Ch10: 595-642 nm

BV605

Ch11: 642-745 nm

AF647, APC

Ch12: 745-800 nm

APC-Cy7, APC-AF750, APCH7

Fluorescence Activated Cell Sorting (FACS)

FIASR has two cell sorters in a BSL2+ facility. One is available for operator-assisted sorting and the other is available for use 24/7 to trained users by reserving time on the FOM calendar.

Operator assisted sorting: Researchers bring stained single cell suspensions to the Resource with controls, buffers, and collection tubes. Together with the operator, the experiment is set up and sort logic is defined. After setup, researchers will not need to be present for the remainder of the sort.

New sorts: Submit the online Sort Information form to request time. The FACS operators will contact you to confirm your appointment and to discuss specifics.

Repeat sorts: Submit the short Sort Appointment Request form to request time. FACS operators will contact you to finalize the appointment details.

Appointments are not guaranteed until confirmed and entered in FOM.

Researchers can refer to “Guidelines to Sorting” for more detailed information.

Contact Information:

Please email Xiaojun Liu (Xiaojun.Liu@RoswellPark.org) or Jennifer Chinnici (Jennifer.Chinnici@RoswellPark.org).

FACS Aria II
  • 4L, 13 fluorescent parameters with 2 scatter parameters (similar to Fortessa B and C)
  • Sheath pressure is adjustable from 2 to 75 psi
  • Nozzle sizes 100 micron (85 and 130 micron also available)
  • Sort options: 4-way tube sort, plate sorts (6, 12, 24, 48, 96, 384 well plates)

Laser

Bandpass Filter

Common Fluors (list is not exhaustive)

355 nm UV laser

U379/28

BUV395, Indo-1

U460/14

DAPI, Hoechst 33342, LD Blue, Zombie UV

405 nm violet laser

V450/50

BV421, BV480, DAPI, Pacific Blue, V450, EF450, SB436, AF405, LD Violet

V530/30

BV510, V500, AmCyan, LD Aqua

488 nm blue laser

B530/30

FITC, BB515, GFP, AF488, CFSE

B575/25

PE, mOrange, AF568

B610/20

PECF594, PEAF610, PEDazzle594, PEeFluor610, ECD, PI

B710/50

PerCPCy5.5, BB700, PerCP, PECy5, PECy5.5, 7AAD, PEeFluor710

B780/60

PECy7, PEAF750

640 nm red laser

R660/20

APC, AF647

R730/30

AF700, APC-R700, DRAQ5

R780/60

APC-Cy7, APC-AF750, APCH7

Sony MA900
  • Available 24/7 to trained users by reserving time on the FOM calendar
  • 4L–colinear for 488/561 nm laser lines and 405/638 nm laser lines
  • 12 fluorescent and 2 scatter parameters
  • Available nozzle sizes: 70, 100, and 130 micron
  • Sort options: 4-way tube sort, plate sorts (6, 12, 24, 48, 96, 384 well plates)

Laser

Bandpass Filter

Common Fluors (list is not exhaustive)

488 nm blue laser
561 nm yellow/green laser

525/50

FITC, BB515, GFP, AF488, CFSE

585/30

PE, mOrange, AF568

617/30

PECF594, PEAF610, PEDazzle594, PEeFluor610, ECD, PI

695/50

PerCPCy5.5, BB700, PerCP, PECy5, PECy5.5, 7AAD, PEeFluor710

785/60

PECy7, PEAF750

405 nm violet laser
640 nm red laser

450/50

BV421, BV480, DAPI, Pacific blue, V450, EF450, SB436, AF405, LD Violet

525/30

BV510, V500, AmCyan, LD Aqua

585/30

BV570

617/30

BV605

665/30

APC, AF647, BV650

720/60

AF700, APC-R700, DRAQ5

785/60

APC-Cy7, APC-AF750, APCH7, BV785, BV786

Mass cytometry analyzer

CyTOF (Standard Biotools) – RETIRING DEC 2024

Helios/CyTOF, a time-of-flight mass cytometry, can acquire single cell suspension labeled with transition metal isotopes-conjugated monoclonal antibodies. The main advantages of mass cytometry over conventional fluorescence-based flow cytometry are minimal spectral overlap of each parameter, thereby, the CyTOF can theoretically detect up to 135 isotopes without compensation.

Since the metal isotopes used for conjugation are rare in biological samples and the environment, it can detect the specific signal of each protein on individual cells regardless of autofluorescence.

This system will be retired in December 2024, no new projects will be accepted. Researchers will be directed to full spectrum flow cytometers as an alternative.

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Slide-based immune profiling/imaging

Zeiss LSM700

First-time users of the Zeiss LSM700 confocal microscope should contact Orla Maguire (Orla.Maguire@RoswellPark.org) to schedule new user training before working independently on the microscope. Operator assistance is available by scheduling an appointment during regular business hours.

The Zeiss laser scanning confocal microscope, equipped with 4 solid-state lasers, is capable of scanning a 512 x 512 pixel region at five frames per second with a zoom factor ranging between 0.5x and 40x. In addition, the unique scanning unit contains a variable dichromatic mirror that enables spectral imaging coupled with linear unmixing to untangle highly overlapping emission spectra.

  • 4 Laser lines: 405nm, 488nm, 555nm, and 639nm
  • Objectives: 10X Air, 20X Air, 40X Oil, 63X Oil
  • 3D imaging
  • Multiple fluorescence and colocalization analyses
  • Emission Fingerprinting
  • FRAP, FLIP, photoactivation and photoconversion
Phenocycler Fusion

The Phenocycler Fusion is an automated immunofluorescence-based analyzer that performs cyclic visualization of 3 immuno-fluorescent analytes at a time. This process can be repeated multiple times and the simultaneous detection of over 100 immunofluorescent probes have been reported. This imaging process is applicable to FFPE and cryosections of human and mouse origin. A catalogue of pre-validated antibodies is available at The Spatial Biology Company | Akoya Biosciences. Custom conjugation of antibodies to targets not available through the catalog is also available. Instrument use is restricted to FIASR personnel only. For more information on the technology or to schedule an experiment please contact Orla Maguire (Orla.Maguire@RoswellPark.org) or Hans Minderman (Hans.Minderman@RoswellPark.org).

 

Hyperion Image Mass Cytometer (IMC) is a high-resolution laser ablation system for 1-micron cellular resolution, allowing the most comprehensive analysis of the immune and non-immune contexture in the tumor microenvironment. The IMC module uses the CyTOF instrument to detect up to 40 markers simultaneously on a tissue slide by measuring the abundance of metal isotopes tagged to antibodies.

This system will be retired in December 2024, no new projects will be accepted. Researchers will be directed to the Phenocycler Fusion or Zeiss confocal microscope as an alternative.

Live cell imaging

xCelligence-MP (Agilent)

The Agilent xCELLigence real-time cell analysis (RTCA) multiple plates (MP) system provides a unique and powerful means to monitor cells in real time, without the potential artifacts generated by using labels. This noninvasive measurement allows detection of changes in adherence, morphology, and viability without the need for overexpression of reporter and target proteins. This provides physiologically relevant data throughout the entire experiment.

This instrument is currently housed in an incubator in Dr. Brentjens department on the 5th floor of the Cancer Cell Center. Training and calendar access requests should be discussed with Dr. Terence Purdon (Terence.Purdon@RoswellPark.org).

Plates used for the instrument are available from FIASR upon request at $66 each. Please email FICSR@RoswellPark.org to arrange pickup.

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Proteomic analyzers

Intelliflex and Luminex 200

The xMAP INTELLIFLEX® System is a flow-bead multiplex analyzer that can perform up to 500 tests in a single reaction volume using magnetic beads to capture soluble analytes from plasma, serum, and tissue culture supernatants.

The Luminex® 200™ System is an older version of the Intelliflex technology that can perform up to 100 tests in a single reaction volume. Multiplexing kits for both instruments are commercially available from several vendors, including Millipore, BioRad, ThermoFisher, and R&D Systems, with assays for many different species and sample types.

New Assay Requests: Contact our Luminex operators to discuss project specifics.

Nitika Sharma: (Nitika.Sharma@RoswellPark.org)
Courtney Ryan: (Courtney.Ryan@RoswellPark.org)

Isolight machine

The Isolight analyzer from Isoplexis/Bruker performs automated highly mutliplexed live single cell secretome analysis, single cell highly multiplexed phosphoproteomic and metabolic analysis as well as bulk/population protein analysis all on one platform and in a single workflow. Fully analyzed data can be obtained within 24 hours. IsoLight is ideal for comprehensive profiling of the secretory capabilities of different cell types and of cells at different developmental or treatment stages. Click on the links to browse the commercially available analyte human and murine panels for bulk (Codeplex) or single cell (Isocode) analysis.

The enzyme-linked immunosorbent spot (ELISpot) quantitatively measures the frequency of cytokine secretion for a single cell. A plate is coated with antibodies for the cytokine of interest. T cells are added with/without stimuli. The coating antibody captures the secreted cytokine and a second cytokine-specific antibody is coupled to a chromogenic substrate and used for detection. Results appear as spots, with each spot corresponding to one cytokine-secreting cell. The number of spots equals the number of cytokine-secreting cells for a specific antigen. The modified FluoroSpot assay uses a fluorescently tagged antibody instead of a biotin-streptavidin substrate system.

FIASR has the CTL S6 FluoroCore Analyzer that can also perform PBMC counting. The FluoroCore is equipped with optics and an extra fluorescent channel which, along with CTL’s dedicated LDA Cell Counting Suite, can count dozens of PBMC samples within minutes, including the documentation of viability, and the calculation of resuspension volumes.

CTL provides several kits for the reader in FIASR (one color kits).

  • 1 laser (488 nm) with 520 bandpass filter (FITC, Alexa Fluor® 488, Acridine Orange, sfGFP, GFP, Yo-Pro™, CFSC, Sytox™ Green)
  • 96- or 384-well plate and Strip plate in 96 well format
  • Single-color FluoroSpot analysis for precise quantification of per-cell secreted analyte via the HDR imaging feature
  • Serves as a high-throughput reader for cell-mediated cytotoxicity assays
  • FCS standard data output
ELLA

The ELLA automated immunoassay system performs immunoassays in a microfluidic Simple Plex cartridge, allowing researchers to acquire highly reproducible validated assay data with very few manual steps, and is the only self-contained testing platform for the simultaneous execution of multiple ELISA-based immunoassays.

Once a sample is loaded, it runs through a microfluidic channel that binds the protein of interest. Next, Ella washes off unbound analyte and adds a detection reagent. Because each channel has three Glass Nano Reactors (GNRs) coated with a capture antibody, you will get triplicate answers for each sample. Answers are then generated from the factory-calibrated standard curve that comes with every cartridge.

The assay volume required is 25 μL or less for up to 4-plex customized assays and the data is ready in two hours from sample prep. Cartridges are available ready-to-use or customizable at Bio-Techne.

Metabolic analyzers

Seahorse XFe96

Seahorse XFe96 extracellular flux analyzer tracks and records the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) that reflects mitochondrial respiration and glycolysis in real-time in 96-well plate format.

Validated assays include the glycolysis stress test, cell mito stress test, mito fuel flex test, glycolytic rate assay, ATP rate assay, cell energy phenotype test and fatty acid oxidation measurement. Custom-made assays are also available. All kits can be viewed at Agilent.

Normalization of Seahorse data can be facilitated by in-situ cell counting using the BioTek Cytation 5, requiring no additional sample processing. Cytation 5 is a multichannel cell imaging platform capable of automated cell counting from fluorescently labeled nuclear images following the injection of cell permeable dye such as Hoechst 33342 via the Seahorse system. The instrument also performs high-contrast brightfield imaging, capturing whole-well images to provide visual feedback and quality control of cell seeding conditions.

calScreener

The Symcel calScreener™ measures the total metabolic activity of a sample, in real-time, and in unlimited run-time that can be set from hours to months. The calScreener directly measures the heat produced as a result of metabolic processes in the sample; the energy output is measured in μW.

The calScreener is a new addition to FIASR (June 2024) and we welcome any studies to test out the instrument. Contact Orla Maguire (Orla.Maguire@RoswellPark.org) or Courtney Ryan (Courtney.Ryan@RoswellPark.org) for more information.

Small particle analyzer

ZetaView

The ZetaView small particle analyzer employs Nanoparticle Tracking Analysis (NTA) which relates the rate of Brownian motion of the particles in liquid suspension to particle size. The ZetaView NTA measures the size distribution profile of small particles approximately 10-1000 nanometers (nm) in suspension. The ZetaView is also capable of measuring the zeta-potential of the particles which is a measure of the electric charge on the nanoparticle surface.

The primary application of the NTA analysis at Roswell Park is measurement of extracellular vesicles (EV) and exosomes, cell-derived vesicles with a diameter between 30 and 100 nm.

The ZetaView analyzers are equipped with either a 488nm or 405nm laser to detect fluorescent particles if desired.

The ZetaView instruments are available 24/7 to trained users by reserving time on the FOM calendar.

Interested researchers should contact our principal operator Nitika Sharma (Nitika.Sharma@RoswellPark.org) or Courtney Ryan (Courtney.Ryan@RoswellPark.org) for project consultation and to discuss full service or training opportunities.

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Analysis software

  • FCS Express 7.0: Roswell Park has a network license free for all Roswell employees. Order software through ServiceNow.
  • FlowJo 10: FIASR has 2 workstations with FlowJo that are available for use during business hours. Individual PIs are responsible for purchasing their own licenses if desired.
  • WinList 9.0: Roswell Park has a network license free for all Roswell employees. Contact Joseph Hanson to arrange access.
  • Modfit 4.0: Roswell Park has a network license free for all Roswell employees. Contact Joseph Hanson to arrange access.
  • OMIQ: Roswell has several single seat licenses available for Roswell employees. Inquiries can be made to Hans Minderman.
  • IDEAS 6.2 (Amnis): FIASR has a license free for all Roswell employees.
  • Amnis AI: Amnis AI works with the IDEAS 6.3 analysis software to help researchers classify objects using computer-aided tagging and AI algorithms. FIASR has one workstation with Amnis AI and IDEAS v6.3, access is available for use during business hours. Inquiries can be made to Hans Minderman.
  • Image Pro Plus 6.3: Image Pro Plus 6.3 is an advanced image analysis software. It accepts a wide variety of image file formats. FIASR has one seat license available for use during business hours.
  • ImageJ: ImageJ is a Java-based free-ware image analysis package developed at the National Institutes of Health. Roswell employees can order the software through ServiceNow.

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New accounts

FIASR uses FOM (Facility Online Manager) as an integrated calendar, instrument sign-on, and billing system. Every new user will need to set up an individual account in FOM.

Each lab (or company) will also need a group account for logging in to the computer software on our analyzer instruments. Only one account will be created per PI. For more information on new accounts, contact Orla Maguire.

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Education

FIASR staff teach several free courses to educate the Roswell Park and Buffalo community on the techniques employed in our shared resource. Classes typically take about one - three hours.

MANDATORY: Introduction to Flow Cytometry: Didactic 1.5hr class introducing the basics of flow cytometry, experimental design, analysis, and outlining the rules to follow when in FIASR.

Three 2-hour one-on-one instrument training sessions include:

  1. Initial training using our validated samples
  2. Researcher brings their own samples
  3. Troubleshooting and reenforcing correct procedures

For those instruments that are available 24/7, an additional training session with FIASR staff is required for after-hours access.

Other optional classes are offered according to demand. Drs. Minderman and Maguire also teach the graduate course RPG564 Principles of Flow and Image Cytometry in the Fall semester open to all graduate students and available to audit is space permits.

Contact Orla Maguire (Orla.Maguire@RoswellPark.org) for more information.

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Roswell Park Comprehensive Cancer Center

Location & hours

Roswell Park Comprehensive Cancer Center
Flow and Immune Analysis Shared Resource
Elm and Carlton Streets
Buffalo, New York 14263

Location: Cancer Cell Center 3rd floor

General lab phone: 716-845-3470
General lab email: FICSR@RoswellPark.org

Grant information

Please acknowledge the Flow and Immune Analysis Shared Resource in your publication as follows:

Cytometry services were provided by the Flow and Immune Analysis Shared Resource at Roswell Park Comprehensive Cancer Center, supported in part by the NCI Cancer Center Support Grant NCI P30CA016056 and NCI R50 R50CA211108.

If you are using the ImageStream imaging flow cytometer in your publications, please acknowledge the Shared Instrument grant that funded the acquisition of this instrument as follows:

The ImagestreamX-MKII instrument was funded through the NIH Shared Instrument Grant S10OD018048.

If you are using the SONY MA900 in your publications, please acknowledge the Shared Instrument grant that funded the acquisition of this instrument as follows:

The SONY MA900 sorter was funded through the NIH Shared Instrument Grant S10OD025183.

The Flow and Immune Analysis Shared Resource (FIASR) at the Roswell Park Comprehensive Cancer Center provides a comprehensive array of cutting- edge technologies on a variety of platforms including flow and image cytometry, proteomics, metabolomics, and spatial profiling techniques in support of Roswell investigators. The FIASR is recognized by the International Society for the Advancement of Cytometry (ISAC) as one of only 14 world-wide Shared Resource Laboratories for operational excellence and adherence to best practices. The Cancer Center has made a strategic commitment to the expansion of immune- and cell-based therapies consequently increasing the demand for laboratory support for translational biomarkers and clinically-based patient monitoring. FIASR provides an exceptional and essential resource for exploring the biology underlying these new therapeutic modalities. The shared resource improves fiscal and operational efficiencies and the rigor and reproducibility of assays due to the application of ISAC approved standardized maintenance, operating and validation protocols ultimately facilitating implementation of these techniques in the clinical realm. In addition to serving Roswell Park and local research institutions, FIASR serves as a core facility for several biopharma and NIH multi-site clinical trials.

Equipment technologies and services

In addition to the data acquisition software for the equipment listed in the Table, the following analysis software packages are available, free of charge to users: WinList and ModFit (Verity), FCSExpress (DeNovo), IDEAS (Amnis); AMNIS-AI; FlowJo (BD Bioscience), OMIQ, VisioPharm and ImagePro (MediaCybernetics). An internal network on a server managed by the Cancer Center’s IT department with expandable storage capacity has been set up for the data management. All investigators within the facility have password-protected access to this network from their desktop PC’s.

Other support equipment include: Cold room, fume hoods, autoclave and glass washing facilities, 3 CO2 incubators, 3 Bioguard laminar flow tissue culture hoods, benchtop centrifuges, refrigerated microfuges, inverted and conventional microscopes with phase contrast, fluorescence capacity and digital camera attachments, PCR, -80 freezers and liquid N2 storage tanks, ELISA reader, and smaller equipment necessary for cellular and molecular studies.

Major Services of the FIASR: (i) Conventional, full spectrum and mass flow cytometry, (ii) Sorting, (iii) Imaging flow cytometry, (iv) Spatial multi-omic analysis, (v) Live cell imaging, (vi) Luminex multiplex bead assays, (vii) Single Cell Secretome analysis, (viii) Nanoparticle sizing, (ix) ELISPOT, (x) ELISA, (xi) Education and training on all of the above, (xii) Sample preparation and analysis for all of the above.