Flow Cytometry Core

Slide-based imaging cell counter

• Brightfield, trypan blue, and AO/PI compatible

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Five-laser, spectral analyzer (16 violet, 14 blue, 10 yellow-green, 8 red)

• Can accept samples in tubes or plates
• Immunophenotyping surface and intracellular antigens
• Viability
• Apoptosis
• Transfection efficiency analysis
• Cell cycle
• Mitochondrial function

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Four-laser, 13 color instrument (4-violet, 2 blue, 4 yellow green, 3 red)

• Can accept samples in tubes or plates
• Immunophenotyping surface and intracellular antigens
• Viability
• Apoptosis
• Transfection efficiency analysis
• Cell cycle
• Mitochondrial function

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A five-laser, 27 color cell sorter in a biosafety cabinet

• Up to six-way tube sorting
• 96-well and 384-well plate sorting
• Single cell and bulk sorting modes
• Side populations
• Immunophenotyping surface and intracellular antigens
• Viability
• Apoptosis
• Cell cycle
• Transfection efficiency analysis
• Staff run

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A four-laser, 12 color cell sorter in a biosafety cabinet

• Up to four-way tube sorting
• 96-well plate sorting
• Immunophenotyping surface and intracellular antigens
• Single cell and bulk sorting modes
• Viability
• Apoptosis
• Cell cycle
• Transfection efficiency analysis
• Staff run

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CBC with five-part differential

• 22 parameters
• Direct eosinophil counts

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Rotor-based chemistry analyzer

• Variety of test menus available

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• All work in the Flow Cytometry Core must be preceded by a consultation with Core Manager Dr. Rachael Sheridan to discuss experimental design, controls, and biosafety.
• Create a CrossLab account. VAI staff that need a login (or further CrossLab support) should contact Lori Moon.

• The Flow Cytometry suite is BSL2.
• No food/drink or containers intended for food/drink in the future are allowed in the space.
• All people in the space must wear long pants (or clothing that covers the legs) and close-toed shoes. Lab coats and safety glasses are recommended.
• Use best practices and universal precautions when handling samples.
• Clean up after yourself and clean up spills in a proper and timely manner. Ask the staff for help if you need it.

• Flow Cytometry Core staff is available from approximately 8:00 a.m.–5:00 p.m. Monday–Friday. If you need access to staff outside of these hours, please make arrangements well in advance. Requests will be accommodated at staff discretion.
• The Core will be closed on Institute-recognized holidays. Users trained for weekend access may still run analytical equipment.
• The CytoFLEX S, Aurora, Luna and VetScan equipment are available 24/7 to well-trained users.
• Researchers are encouraged to seek training to run these machines independently. Training must be performed by Core staff. Trained users may not train others on the instrumentation and have it count as training.

• Consultation with Core staff is required before booking your first sort or analysis run. Consultations may be arranged by emailing [email protected].
• Appointments are booked on a first-come, first-served basis through the CrossLab system. Core staff reserve the right to make small alterations in the schedule to accommodate as many researchers as possible on high-use instrumentation. You will be notified if any changes need to be made to your appointment.
• Do not share accounts. The name of the person booking the appointment must match the identity of the user at the instrument. Violators may have their accounts deactivated.
• Billing time begins at the beginning of your scheduled appointment or when you log in to the cytometer, whichever is earlier.
• Billing time ends at the end of your scheduled appointment or when you log out of the cytometer, whichever is later.
• If you are more than 15 minutes late for your appointment without notifying Core staff of your updated timeline, you may forfeit your appointment at staff discretion and will be allowed to run later only if schedules allow.
• Cancellations with less than 24-hour notice will be billed in their entirety unless the time is rebooked by another user. Exceptions for unforeseeable circumstances may be granted at the staff’s discretion. Cancellations may be made through the CrossLab website (more than 24 hours in advance) or by contacting Core staff by email.

• Users are responsible for annotating, transferring, and maintaining data generated in the Flow Cytometry Core.
• Annotate your data at the instrument as much as possible.
• The Core will maintain data on the instrument computers for one month after which it will be deleted. Templates will remain available indefinitely.

• Cytek Bio Resources-Cytek Bio
• Introduction to Flow Cytometry–BD Biosciences
• Flourescence Tutorials–ThermoFisher Scientific
• Multicolor Flow Cytometry: The Brilliant Violet Family of Fluourophores–BioLegend

• Flow Cytometry–A Basic Introduction–DeNovo Software
• The Molecular Probes Handbook–ThermoFisher Scientific
• Compensation Beads–UWCCC Flow Lab
• Rainbow Bead Standardization–UWCCC Flow Lab
• Single Cell Sorting for Transfected Cells–UWCCC Flow Lab
• Multicolor Flow from Start to Finish: A Quick Reference Guide–UWCCC Flow Lab
• Minimizing Aggregates in Samples–UWCCC Flow Lab
• Titrating Antibodies for Flow Cytometry–UWCCC Flow Lab
• FlowJo for Antibody Titrations: Separation Index and Concentration–UWCCC Flow Lab
• Calcium Flux Protocol–UWCCC Flow Lab
• BD FACSelect Buffer Compatibility Resource–Cytobank

• Cytometry Part A–Wiley Online Library

• Cyto U–International Society for the Advancement of Cytometry
• Expert Cytometry Blog–Expert Cytometry
• International Society for the Advancement of Cytometry
• International Clinical Cytometry Society
• Purdue University Cytometry Laboratories
• Chromocyte

• BenchSci A.I. Assisted Antibody Search
• FluoroFinder
• Guided Panel Solution – BD
• BioLegend Essential Markers – BioLegend

• Fluorescence SpectraViewer–ThermoFisher Scientific
• BD Fluorescence Spectrum Viewer–BD Biosciences
• Fluorescence Spectra Analyzer–BioLegend
• Chroma Spectra Viewer–Chroma

• Flow Cytometry–BD Biosciences
• Flow Cytometry–Affeymetrix eBioscience
• BioLegend
• Molecular Probes–ThermoFisher Scientific
• Tonbo Biosciences
• Miltenyi Biotec
• Spherotech