Flow Cytometry Core

OVERVIEW

The Flow Cytometry Core is dedicated to providing high-quality, comprehensive cytometry services and expertise to scientists at the Institute and external collaborators. The Core’s instrumentation includes a 21-color MoFlo Astrios cell sorter, a 14-color CytoFLEX S benchtop analyzer and a 4-laser spectral Aurora cytometer; a GentleMACS Octo tissue dissociator with heaters; a Shandon Cytospin 3; a VetScan HMII for CBCs; and a VetScan Classic for blood chemistry analysis.

How to Request Services

VAI’s Core Technologies and Services manages service requests through CrossLab Solutions. Each Core maintains a CrossLab page that provides descriptions of the Core as well as options to schedule equipment and request services. Pricing also is available in CrossLab (See Schedule Equipment and/or Request Services). Current CrossLab users can login here.

VAI staff that need a login (or further CrossLab support) should contact Lori Moon. MSU and external users that need a CrossLab account/login should go here for account creation. After logging in, external users should click “list all cores” under the Core Facilities section in the left menu and scroll down to “Cores at Van Andel Institute.”

For questions about the Core, please contact Flow Cytometry Core Manager Rachael Sheridan, Ph.D., at [email protected].

Our Impact

We’re raising thousands to save millions.

We’re turning hope into action for the millions of people around the world affected by diseases like cancer and Parkinson’s. Find out how you can help us make a difference.

  • 118 papers published in 2022
  • 48 papers in high-impact journals in 2022
  • 42 clinical trials launched

Instrumentation

5-laser, 21-color cell sorter (3 UV, 6 violet, 3 blue, 5 yellow-green, 3 red) in a biosafety cabinet

  • Up to six-way tube sorting
  • Plate sorting up to 96-well plates
  • Side populations
  • Immunophenotyping surface and intracellular antigens
  • Single-cell sorting for subcloning or genetic analysis
  • Viability
  • Apoptosis
  • Cell cycle
  • Transfection efficiency analysis
  • Mitochondrial function
  • Primarily staff-run, semi-assisted option for well-trained users

Read more here.

Four-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
  • Staff- or self-run
  • Available 24/7 to well-trained users

Read more here.

Four-laser, 14-color benchtop analyzer (4 violet, 2 blue, 4 yellow-green, 4 red)

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

Read more here.

Blood chemistry analysis

  • Comprehensive Diagnostic Profile
  • Prep Profile II
  • Critical Care Plus
  • T4/Cholesterol Profiles
  • Mammalian Liver Profile
  • Equine Profile Plus
  • Avian/Reptilian Profile Plus
  • Large Animal Profile
  • Available 24/7 to well-trained users

18-parameter hematology analyzer

  • CBC with three-part differential (Lymph, Mono, Gran)
  • Hemoglobin and hematocrit
  • Mean corpuscular volume
  • Mean corpuscular hemoglobin
  • Platelet volume
  • Available 24/7 to well-trained users

Gently deposit a monolayer of cells on your slide

  • Core provides reusable rotors and disposable funnels with integrated cards
  • Spin up to 24 samples
  • Speeds from 200 to 2,000 rpm
  • Available 24/7 to well-trained users

Services

  • Experimental design
  • Instrumentation choice
  • Panel design and fluorochrome choice
  • Panel optimization methods
  • Sample preparation
  • Biosafety
  • Special sort considerations
  • Cytexpert
  • FlowJo v10
  • Kaluza
  • Summit
  • Available 24/7 to trained users
  • Assistance with grant preparation
  • Assistance with manuscript preparation
  • Production of publication-quality graphics

Policies & Procedures

  • 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 and MSU staff that need a login should contact Erica Lange. External users that need a CrossLab account/login can go here for account creation. After logging in, external users should click “list all cores” under the Core Facilities section in the left menu and scroll down to “Cores at Van Andel Institute”.
  • 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, Cytospin 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.
  • It is strongly recommended that users attend a section of Flow Cytometry 101 before training.
  • Training is initiated by contacting the Core staff directly.
  • The initial training appointment will be performed with beads provided by the Core to acclimate the user to the software and instrument operation.
  • A second training appointment with the user’s own samples is also required to evaluate user’s abilities to prepare samples and run the instrument. Core staff will be available for the duration of this appointment to provide guidance and answer questions.
  • After the second training, users may operate independently during business hours and are encouraged to consult with staff if any questions arise.
  • When the user is comfortable and has demonstrated proficiency with the instrument, they may have 24/7 access.
  • Trained users assume responsibility for instrument operation and for performing basic actions to maintain serviceability.
  • Independent users are responsible for ensuring their samples are properly prepared and filtered.
  • Trained users are responsible for ensuring that their data is properly transferred after their run.
  • Independent users are responsible for cleaning the instrumentation and workspace in a manner appropriate for their samples at the end of their appointment.
  • Users must notify Core staff of any problems or abnormalities encountered during their run.
  • Flow Cytometry Core staff will ensure the sorter is ready and aimed for your sort in accordance with your CrossLab appointment form.
  • The user is responsible for knowing how to open/create protocols, regions, sort logic, etc. in Summit software.
  • The user is responsible for appropriate sample preparation. The Core provides sterile filters for use as needed. Users must filter all samples immediately before loading them on the Astrios.
  • The user is responsible for appropriately cleaning the Astrios according to the laminated cleaning and shut down protocol by the instrument. The last user of the day is responsible for instrument shut down.
  • The user is responsible for cleaning up their work areas in the sort room, and around the cytometer and cytometer computer.
  • The user is responsible for notifying Core staff when the run is complete. Failure to do so will result in billing end time being when the staff found the sorter abandoned.
  • During staffed hours, the user must notify staff of any problems or anomalies encountered during the run.
  • The sorter may not be left unattended with less than 500 mL in the sample tube for a stable sort. The sorter may never be left unattended for a sort with unstable event rate or a history of adverse events. The sorter may not be left unattended for more than 10 minutes. Please ask staff to step in and cover for you as needed.
  • The trained person who makes the appointment must be the person who is at the sorter. Untrained users from the same (or other) labs at the sorter in the absence of the trained user will count as an unattended sorter.
  • The sorter is a sought-after commodity and we sometimes need to accommodate unscheduled or shifted-schedule runs. If you are more than 15 minutes late for your appointment without notifying the Core of your updated timeline, you may forfeit your appointment at the discretion of the Core. You will still be charged for your setup fee.
  • 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.

References

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

Rachael Sheridan, Ph.D., SCYM(ASCP)

Director, Flow Cytometry Core

Madison Nichols, B.S, MLS(ASCP)CM

Flow Cytometry Specialist I, Flow Cytometry Core

Josh Schipper, Ph.D.

Flow Cytometry Specialist I, Flow Cytometry Core