Flow Cytometry Core Facility

As one of the University of Toledo's core laboratories, the Flow Cytometry Core Facility provides access to instrumentation, assistance, and guidance, and offer information on current cytometric methods and applications as well as on dyes, cytometric bead arrays, multi-color panel kits, and other reagents. We welcoming UT investigators from all campuses and industrial researchers, technicians, and students from the surrounding community.                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

In addition the Flow Core also offers high-speed cell sorting. This can be used to isolate/separate cell types (or vesicular bodies), rare populations, as well as specific clones post-transfection for further cell culture or analysis/experimentation—if a population can be labeled, distinguished, and gated, it can be sorted.

Samples are prepared by individual investigators who then deliver them to the Core for flow cytometric analysis or sorting. The core will not prepare cells but can provide assistance, guidance, and/or generation of publication-quality graphics and experimental design.

 

The Core Facility provides the following equipment:

BD FACSDiscover™ S8 Cell Sorter

Features BD CellView™ Image Technology and BD SpectralFX™ Technology to expand the power of cell analysis and sorting by combining flow cytometry data with spatial and morphological insights.

Optics

Lasers

Excitation optics

• Violet - 405 nm - nominal 50 mW; output power: >45 mW
• Blue - 488 nm - nominal 100 mW; output power: >95 mW
• Yellow/Green - 561 nm - nominal 50 mW; output power: >45 mW
• Red - 637 nm - nominal 100 mW; output power: >90 mW
• *488-nm laser is optically divided to support signal detection with BD CellView™ Image Technology

Scatter detectors

• Blue laser:  Forward side scatter (FSC), Side scatter (SSC), Axial light loss (ALL)
• Violet laser:  Axial light loss (ALL), Side scatter (SSC) Fluorescence detectors for spectral flow cytometry

Fluorescence detectors

Spectral arrays – 56 fluorescence APD detectors paired with algorithmically optimized filter bandwidths covering the full spectrum:

• Violet 405-nm laser – 20 Violet detectors, covering 410–860 nm
• Blue 488-nm laser – 16 Blue detectors, covering 495–860 nm
• Y/G 561-nm laser – 12 Yellow-green detectors, covering 570–860 nm
• Red 637-nm laser – 8 Red detectors, covering 645–860nm

Imaging optics

Image-enabled detectors

Blue laser scatter detectors

• Forward scatter (FSC), Side scatter (SSC), Axial light loss (ALL)

Fluorescence detectors for imaging (PMTs):

FL1: LP505: 534/46 FL2: LP570: 600/60 FL3: LP675: 788/225

• Class II Type A2 biological safety cabinet (BSC)
• Aggregate Discrimination by BD Doublet Discrimination Module
• Multi-well Plate Sorter
• Temperature Controlled Sample and Sorting Receptacles

Sort

Nozzles, pressure and frequency

• 85-μm nozzle: 35 psi, up to 6-way sorting
• 100-μm nozzle: 20 psi, up to 4-way sorting
• 130-μm nozzle: 7 psi, up to 2-way sorting

Sort collection devices

Tube sorting:

• 1.5-mL microtubes, 2.0-mL tubes and 5.0-mL 12x75-mm tubes

Plate sorting:

• 6-, 24-, 48-, 96- and 384- well plates, microscope slide (optional)

Index sorting:

• Correlation of imaging features and traditional spectral flow cytometry parameters of sorted events with well location of a multi-well plate.

 

BD FACSCanto™ II Cell Analyzer

Optics

Lasers

Excitation optics

• Blue - 488 nm - nominal 100 mW; output power: >95 mW
• Red - 637 nm - nominal 100 mW; output power: >90 mW

Scatter and Fluorescence detectors

Blue laser:

• 488nm Excitation, Forward side scatter (FSC), Side scatter (SSC), Side scatter (SSC) Fluorescence detectors for spectral flow cytometry
• SSC- 488/10
• 530/30- 515 to 545nm Emission
• 575/26- 562 to 588nm Emission
• 610/20- 600 to 620nm Emission
• 695/40- 675 to 715nm Emission
• 80/60- 750 to 810nm Emission

Red laser:

• 633nm Excitation
• 660/20- 650 to 670nm Emission
• 730/45- 707 to 752nm Emission

More information is available at BD Biosciences website

Services

Hours of operation, fees, and scheduling

The Flow Cytometry Core Facility will be staffed from 8:30 a.m. until 4:00 p.m., Monday through Friday.

Any inquiries or scheduling can be made by phone at 419.383.3402 or by email.

The Core Facility operates on a Fee-for-Service basis which will include data acquisition, analysis and consultation if needed.

Current Flow Rates
	Assisted	Self-Operation
FACSCanto II	$60/hr	$60/hr
FACSDiscover S8	$125/hr	$100/hr

Cancellations with less than a 24-hour notice are billed for the scheduled time. Investigators are not billed for cancellations with at least 24 hour notice. Exceptions will be determined by the Core Facility.

The Core Facility is located in the Health Education Building room 233A on the University of Toledo Health Science Campus.

 

Located on the second floor of the Health Education Building in Room 233A, the lab is directly accessible from the second floor hallway of the Pharmacy Building and Dowling Hall.

 

Visitor Parking

For visitors coming from off campus, please follow this link to purchase day parking passes from ParkUToledo.

Parking is located in the front of the Health Education Building in Area 43. Alternate parking is available in Areas 44, 46 (not pictured above), and 48 with access to Health Education through the adjoining buildings or basement tunnel.

A full campus map is available for download.

Flow basics click here