Flow cytometry

A graphic showing a machine with coloured light coming out in rays.

Services we offer

Flow cytometry cell analysis

Flow cytometry is a laser-based technology that allows for simultaneous multi-parametric analysis of physical and/or chemical characteristics of cells at the single-cell level.

Fluorescence-activated cell sorting

Fluorescence-activated cell sorting is a specialized type of flow cytometry where a heterogeneous mixture of cell suspension can be sorted into up to four populations of interest based on specific light scattering and fluorescent characteristics.

Fluorescence microscopy

A fluorescence microscope is an optical microscope that uses fluorescence and phosphorescence instead of, or in addition to, reflection and absorption to study properties of organic or inorganic substances.

FlowJo site licence program

A FlowJo Portal site license is a user-based license management system for a group or institution. FlowJo Portal provides an improved management interface to site administrators.

Immunophenotyping

Immunophenotyping is a technique used to study the protein expressed by cells.

Training

To receive access to equipment, you must first become familiar with the basic operational procedures within the facility and complete a training session.

The University of Manitoba Flow Cytometry Core Facility is a multi-user facility that provides a wide range of flow cytometry-based services. Our state-of-the-art flow cytometry analyzers and sorter offer researchers the capability to design and perform multicolour assays that best suits your needs with high efficiency and high quality.

Flow cytometry training

We provide individual one-on-one training that is specifically designed to the needs of your project on our flow cytometry analyzers. By taking our training course, you will not only gain a better understanding of flow cytometry in general but also perform sample acquisition and data analysis on your own shortly after the training.

Fees

Training for LSRFortessa, Cytoflex LX, and FACSCanto-II

Training is available for both new and experienced users of flow cytometry:

New users: Two sessions
- 2-hour introductory lecture
- 2-hour hands-on supervised sample acquisition
Experienced users: One session
- 30-minute background overview
- 1.5-hour hands-on supervised sample acquisition

Training fees

User type	User level	Session details	Fee
Academic	New user	2.5-hour lecture 2.5-hour hands-on	$175 (lecture) $175 (hands-on)
Academic	Experienced user	2-hour hands-on session	$175
Industry	New user	2.5-hour lecture 2.5-hour hands-on	$350 (lecture) $350 (hands-on)
Industry	Experienced user	2-hour hands-on session	$350

We’re committed to supporting all trainees until they feel fully confident operating the equipment independently. Additional hands-on or refresher training is available on request at $70/hour.

New users

Training for new users consists of two introductory sessions.

Session 1 (two hours)

Presentation on the basics of flow cytometry, including:

compensation
experimental setup
fluorescence basics and excitation/emission spectra
multicolor panel design
plots and gating
sample isolation and preparation

Session 2 (two hours)

Hands-on practice, with supervision. This will include instrumental setup and calibration, sample acquisition and data analysis on the flow cytometry analyzer on your own cell samples.

Prepared samples are required.

Experienced users

Training for experienced users consists of one two-hour session, consisting of:

Presentation on the basics of flow cytometry, including: fluorescence spectral overlap, instrument review and data analysis. (30 minutes)

Hands-on practice, with supervision. This will include instrumental setup and calibration, sample acquisition and data analysis on the flow cytometry analyzer on your own cell samples. (60 – 90 minutes)

Prepared samples are required.

Prepared samples

To complete the training, both new and experienced users must bring prepared samples. Using your own samples will allow you to quickly learn how to acquire data in future experiments.

Samples generally consist of:

One or two tubes of unstained samples
Single stained controls of all the fluorochrome you are using if you are doing multi-colour staining
Four to 10 experimental samples

There should be minimal 0.5 million cells in 200-300μL of PBS in standard flow tubes (12x75mm, 5mL Polystyrene tube).

We recommend you to prepare your own cell samples that will be used in your project in order for you to perform acquisition/analysis on your own as quickly as possible.

You are advised to develop clear project objectives involving flow cytometry prior to training in order obtain the greatest benefit from the course.

Support

We continue to provide support and assistance to the users of our facility after training on an as-needed basis.

Upon request, we can also provide assisted sample acquisition/data analysis during real experiments to show competency in independent flow cytometry acquisition capability.

Equipment

The following equipment is available for use within our facility:

Guidelines

Acknowledgments

If research supported by the University of Manitoba Flow Cytometry Core Facility results in publication, we would appreciate the following acknowledgment in your publication(s):

"We thank the Flow Cytometry Core Facility at the University of Manitoba in Winnipeg, MB, for providing __________ service."

Biosafety

All flow cytometry instruments in the facility are housed at bio-safety level 2 (BSL2) labs. Therefore, only live samples with BSL no higher than level 2 are allowed to enter the facility. Samples from risk group BSL3 or higher must be pre-treated with paraformaldehyde before accessing the facility and the facility manager must be informed in advance. The user need to ensure that enough bleach is present in the waste tank of the instrument during acquisition when using the flow cytometry analyzers (FACSCanto and LSR-II).

The users must have their formal bio-safety training completed and lab permits available before using the facility. The facility manager reserves the rights to:

request for clarifications and BSL documents from the facility users when necessary, and/or
reject the entrance of certain samples from the users if bio-safety regulations are not adhered to.

The FACSAria-III is housed in a Class II type A2 bio-safety cabinet to ensure proper aerosol management of the sample during sorting.

Contact Dr. Christine Zhang for details if you have further questions or concerns.

Data management

Good data management practice is vital to keeping facility instruments running and problem-free. You are strongly advised to back up your data onto a flash drives as data left on facility workstations will be deleted regularly.

Notices will be sent out to remind users about this practise and to provide a date on which the deletions will take place. The facility is not responsible for loss of data due to failure to back up information by the stated deadline.

Follow these steps to back up your data:

Export your files on under the "Browser" tab of the Diva software as FCS files into "BD export folder" in D: drive.

Then back up your files into your own USB stick or external hard drive H: drive (make sure your USB drive is virus-free).

Finally, delete your files in "D:\BD export"

Delete your experimental files under the "Browser" tab in the Diva software.

Resources

Panel design tool

Designing a multi-color flow cytometry panel can be a daunting and scary task. The following are two population multi color flow cytometry panel design tools you can use that can make your panel sign process much easier and faster.

FluoroFinder: A popular online panel design tool that can help you with multi-color panel design, whether it is using conventional flow cytometers or spectral flow cytometers. Visit the FluoroFinder webpage of our flow cytometry core platform.

BD Research Cloud: BD's online tool with various features including panel designs, data sharing, BD reagents searching, projects organization.

Cytek Cloud: a multi-color designing tool specifically design to streamline panel design for spectral flow cytometry analyzer Aurora.

Feel Free to contact Dr. Christine Zhang if you have any questions.

Fluorescence spectrum viewer

BD Fluorescence Spectrum Viewer

This is an online tool that you can use to visualize the excitation/emission wavelength range of multiple fluorochromes of your choice in comparison to the bandpass filter settings of your instruments.

You’ll be able to get answers to questions like:

Is my instrument properly optimized to detect the fluorochrome(s) I plan to use?
Will there be significant signal overlap or cross-talk between the fluorochromes I’ve selected?

Fluorescence Protein Spectrum Viewer

BD spectrum viewer as well as many other company's spectrum viewer do not include fluorescence protein in their list. Here is a great tool for fluorescence protein spectrum. It is interactive and you will be able to scroll around the excitation and emission spectra to see the exact excitation rate at any given laser wavelength.

Java is required to access both of these tools.