Flow Cytometry Core

Staff

Arti Ghosh

Flow Cytometry Core
Associate Scientist
Phone: (302) 831-0627
aghos@udel.edu
Office: CBBI 244


 

Located on the University of Delaware’s main campus in the Center for Biomedical and Brain Imaging (CBBI) and on the STAR campus at AP BioPharma, the UD Flow Cytometry Core Facility offers a full complement of flow cytometry and cell sorting services:

  • Immunophenotyping
  • DNA Content Analysis
  • Veterinary Hematology
  • Sample Processing
  • Figure Preparation
  • Data Analysis
  • Consultation
  • Assay Design
  • Cell Culture
  • Cell Sorting

We also offer training, workshops and seminar events that increase skills and knowledge about cytometry and cell biology.

BD FACSAria Fusion High Speed Cell Sorter at CBBI

This state-of-the-art high speed cell sorter features:

  • Sterile Biosafety Level II Sorting within HEPA filtered safety cabinet
  • 4 laser lines: 488nm Blue, 405nm Violet, 561nm Yellow/Green, 640nm Red
  • Detection of up to 15 colors in addition to FSC + SSC
  • Sort 4 bulk populations simultaneously or single cell populations into 96/364-well plates
  • 98% live cell purity under ideal conditions (viability dye use encouraged)
  • 3 configurations: 70um nozzle at 70psi (40 million cells/hour), 85um nozzle at 45psi (25 million cells/hour), or 100um nozzle at 20psi (10-15 million cells/hour)

*Please check with core facility staff to make sure your sample can be sorted on the instrument. There is a nominal half hour charge associated with performing aseptic sorting.*

BD Accuri C6 Plus Flow Cytometer

The Accuri C6 is a walk-up, easy to use, 4-color bench top flow cytometer.

  • The system is equipped with a blue laser and a red laser, two scatter detectors and four fluorescence detectors with interference filters optimized for the detection of FITC, PE, PerCP-Cy™5.5 and APC.
  • A compact optical design, fixed alignment and pre-optimized detector settings.
  • A unique low-pressure pumping system drives the fluidics.
  • A sheath-focused core enables event rates of up to 10,000 events per second and a sample concentration over 5 x 106 cells per mL.
  • BD Accuri C6 Plus software has an intuitive user interface.
  • The tabbed interface provides quick access to the collection, analysis and statistics functions.
  • Analysis can be performed on the BD Accuri C6 Plus flow cytometer or can be exported into third-party programs.
  • Walk up use is permitted and encouraged.

BD FACSAria Fusion High Speed Cell Sorter (AP BioPharma)

This state-of-the-art high speed cell sorter features:

  • Sterile Biosafety Level II Sorting within HEPA-filtered safety cabinet
  • 4 laser lines: 488nm Blue, 405nm Violet, 561nm Yellow/Green, 640nm Red
  • Detection of up to 15 colors in addition to FSC + SSC
  • Sort 4 bulk populations simultaneously or single cell populations into 96/364-well plates
  • 98% live cell purity under ideal conditions (viability dye use encouraged)
  • 3 configurations: 70um nozzle at 70psi (40 million cells/hour), 85um nozzle at 45psi (25 million cells/hour), or 100um nozzle at 20psi (10-15 million cells/hour)

*Please check with core facility staff to make sure your sample can be sorted on the instrument. There is a nominal half hour charge associated with performing aseptic sorting.*

VetScan HM5c Hematology Analyzer

The VETSCAN HM5c is a fully-automated, five-part differential hematology analyzer displaying a comprehensive 22-parameter complete blood count (CBC) with cellular histograms on an easy-to-read touch-screen.

Measures WBC (% and absolute count for Lymphs, Monos, Neut, Eos, Baso), RBC (HGB, HCT, MCV, MCH, MCHC), PLT

For use with murine (rodent), canine, equine, bovine, feline (15+ additional species) veterinary blood, marrow, acites and CSF samples (including NHP*)

Can also be used to perform automated counting of cell suspensions from culture or dissociated tissue

*Staff must be informed if you intend to use NHP blood, only approved BSL 2 work is allowed under any circumstances.*

Thermo Scientific Cytospin4

The Thermo Scientific Cytospin4 provides economical thin-layer preparations from any liquid matrix, especially hypocellular fluids such as spinal fluid, needle aspirates and urine.

Provides a good representation of all cell types present in homogeneous liquid samples, especially immunological and hematologic malignancy (blood, bone marrow, PBMCs, pleural effusion).

Deposits a thin layer of cells on slides while maintaining cellular integrity for morphology and pathology evaluation.

Nexcelom Cellometer Vision CBA Image Cytometry System

All-in-One SystemBasic cell counting, primary cell viability, and cell-based assays.

Dual-Fluorescence for accurate primary cell viability (no interference from red blood cells)

Analyze bone marrow, peripheral blood, and cord blood without lysing

Unique algorithms for advanced cell analysis

Determine concentration and viability of hepatocytes, adipocytes, and other sophisticated cell types

Fast results: Obtain cell images, counts, size measurements, viability calculations, and population data in <3 minutes.


 

Additional amenities at the Flow Cytometry Core Facility include:

  • Common FACS / Flow Cytometry Reagents
  • Laminar Flow Hood / BSC Level II
  • Sample Prep / Wet Lab
  • Refrigerators, Freezer
  • Cell Culture Suite
  • Centrifuges
  • Ice Maker

 

Dedicated computer workstations allow users to analyze data in the lab. Software packages include:

  • FACSDiva 8.0.3
  • CFlow Plus / CFlow Sampler
  • FCS Express 5.0
  • CellQuest Pro 6.0.2
  • ModFit LT 2.0

 

Zeiss Axiovert 200

The Zeiss Axiovert 200 is an inverted epi-fluorescence microscope. Images can be aquired with the AmScope MU-1000 USB 3.0 and AmScope Software or Hamamatsu ORCA-ER digital camera and AxioVision software.  The ORCA-ER digital camera is a high resolution gray scale camera.

Standard objectives on this system include:

  • 10x Plan Apochromat, 0.45, Ph1
  • 20x Plan Apochromat, 0.8, Ph2
  • 40x C-Apochromat, 1.2W, oil, DIC
  • 63x Plan-Apochromat, 1.4, oil, DIC

Also located in the Multimodal Imaging Suite at CBBI:

Zeiss LSM980 Confocal / Spectral / Multiphoton

The Zeiss LSM 980 is an upright confocal microscope (i.e. objectives are above the sample) that can be used for confocal, spectral, and multiphoton imaging.

Applications:

  • Confocal imaging.
  • Spectral imaging with linear unmixing. It can be used to remove background autofluorescence or for multicolor imaging when spectral overlaps between colors cannot be separated with standard confocal microscopy.
  • Multiphoton excitation microscopy. It can be used to image up to 10x deeper than confocal microscopy. Two non-descanned PMT detectors can be used for imaging multiphoton fluorescence or second harmonic generation (SHG) in the forward and reverse directions. SHG is now widely used to image collagen in many different applications, and is becoming useful for imaging myosin and some polysaccharides.
  • Confocal reflection microscopy. It can be used to image collagen fibers and surface microtopography.
  • FRET, FRAP, FLIP.
  • Fluorescence Correlation Spectroscopy imaging. It provides quantitative data on concentrations, diffusion coefficients, molecular transport and interactions.
  • Tile scanning to image a large field at high resolution.

Objectives: 10x/0.3, 20x/0.75, 40x/1.2 W, 40x/1.4 Oil – more objectives are available as add-ons.

Laser lines: 405nm, 488nm, 561nm, 639nm, 690-1040nm.

Detectors: a 32-channel ultra-sensitive spectral GaAsP PMT (Gallium Arsenide Phosphide PhotoMultiplier Tube) detector (i.e. ChS in Zen) which is ideal for spectral imaging, two GaAsP PMT detectors (Ch1 & Ch2), one transmitted PMT (T-PMT) for contrast imaging (Bright Field, Oblique illumination, Differential Interference Contrast), and two non-descanned detectors (one GaAsP and one multi-alkali) for multiphoton imaging.

Operating software: Zen 3.5

Location: CBBI 244C

Contact person: Dr. Sylvain Le Marchand