Facility
Flow Facility Sites
- Available instruments:
- Analysers: BC CytoFlex, BD LSRFortessa, Cytek Aurora
- Sorter: AriaIII
- Facility Operator: Hong Liang, liangh@mcmaster.ca
- Ext: 21589
- Available instruments:
- Sorter and operator-assisted analyzer: Beckman Coulter MoFlo XDP
- Facility Operator: Minomi Subapanditha, subapamk@mcmaster.ca
- Ext: 26382
- Available instruments:
- Analyzers: BC CytoFlex LX, BC CytoFlex S
- Sorter: BD AriaIII
- Facility Operator: Zoya Tabunshchyk, zoyatab@mcmaster.ca
- 289-208-2311
Information
Analytical flow cytometers (BC CytoFlex LX, BD Fortessa, Cytek Aurora and Miltenyi MAcsQuant) are available to trained users (with passwords) 24 hours per day, 7 days a week.
- Sorting time has to be arranged with the facility’s operator.
- The facility operator is only available during normal university work hours for assistance.
Effective as of February 1st, 2019
Analysers | Unassisted/hr | Assisted/hr |
BC CytoFlex LX (MDCL 4098) | sliding scale | 80 |
BC CytoFlex LX (MDCL 5144) | sliding scale | 80 |
BC CytoFlex S(MDCL 5144) | sliding scale | 80 |
BD Fortessa (MDCL4098) | sliding scale | 80 |
Cytek Aurora (MDCL4098) | sliding scale | 80 |
MoFlo (MDCL 5058) | N/A | 80 |
Sliding scale rate* | Per Hour |
1-120 hours | $70 |
121-240 hours | $50 |
241+ hours | $30 |
*credits generated starting May 1, 2019 |
Sorters | Unassisted/hr | Assisted/hr |
AriaIII (MDCL4098) | N/A | 80 |
AriaIII (MDCL5144) | N/A | 80 |
MoFlo (MDCL5058) | N/A | 80 |
Training | $80/hr |
Consultation (Data Analysis, Experiment Design) | $50/hr |
FlowJo Site License Fee | ~$350/year |
Instruments
MDCL 4098
Instruments in this Room
Details
- Laser Excitations: Near UV (375nm), Volet (405nm), Blue (488nm), Yellow Green (561nm), Red (635nm)
- Colours: 19
- Functions: high efficiency and low-noise Fiber Array Photodiode Detectors (FAPDs) for superior resolution and sensitivity with 7 decades tunable dynamic range of detection;
- an additional side scatter (VSSC) from the violet (405 nm) laser allows the detection of nanoparticles (<200 nm);
- robust and simple plate loading for 96-well plates;
- volumetric measurements with continuous flow, so no need for counting beads for particle enumeration;
- intuitive, easy-to-operate CytExpert acquisition/analysis software;
- compensation values are automatically adjusted in real-time if the gain settings are changed using a compensation library (a repository of compensation spillover values of dyes) due to the linear response across the entire range.
Configurations
Lasers | Filters | Common Fluorochromes and Dyes |
NearUV (375nm) | 450/45 | NUV450, DAPI, Filipin, Hoechst33342,Hoechst Blue, Hoechst33258, Hoechst34580 |
585/15 | NUV585, BV570, Qdot585, FVS575V, Pacific Orange | |
675/30 | NUV675, BUV661, HoechstRed, Qdot655 | |
Violet (405nm) | 450/45 | V450-PB, BV421, Pacific Blue, eFluor450, V450, Cascade Blue |
525/40 | V525-KrC, BV510, AmCyan, V525, Qdot525, FVS510, AF430, KromeOrange | |
610/20 | V610-BV605, BV605, Qdot605, V610, SuperBright600 | |
712/25 | V712-BV711, BV711, Qdot705, V712, SuperBright702 | |
780/60 | V780-BV785, BV785, Qdot800, V780 | |
Blue (488nm) | 525/40BP | FITC, AF488, GFP, ZombieGreen, CFSE, B525, B525-FITC, Fluo-4 |
675/13BP | PerCP, B675, B675-PerCP | |
712/25BP | PerCP-Cy5.5, B712, BB700, PerCP-eflior710 | |
Yellow Green (561nm) | 585/15 | PE, Y585-PE, Dtomato, DsRed, Y585 |
610/20 | Y610-mCherry, PE-CF594, ECD, PE-Dazzle594, PI, PE-TexasRed, mCherry, Y610 | |
660/10 | PE-Cy5, mPlum, 7AAD, Y660-PC5, Y660 | |
710/50 | PE-Cy5.5, Y710, PE-AF700 | |
780/60 | Y780-PC7, PE-Cy7, Y780 | |
Red (635nm) | 670/30 | APC, Cy5, R670, AF647 |
712/25 | APC-Cy5.5, AF700, APC-R700, R712 | |
780/60 | APC-Cy7, APC-H7, R780-APC-Cy7, APC-AF750, ZombieNIR |
Details
- Laser Excitations: Near UV (375nm), Blue (488nm), Red (633nm)
- Colours: 9 (Flexible configurations are available for specific experiments)
- Functions: Sorter; 4-way sorting into microtubes and 12x75mm tubes,
- Two-way sorting into microtubes, 12x75mm, and 15mL tubes;
- One-way sorting (including single cell) into 6-, 12-, 24-, 48-, 96-, 384-well plates and slides.
Configurations
Lasers | Filters | Common Fluorochromes and Dyes |
Near UV(375nm) | 450/40 | BV 421, DAPI, Hoechst 33342 blue, Pacific Blue, Cascade Blue, Alexa 405, V450 |
670LP | Hoechst 33342 Red | |
Blue (488nm) | 530/30 | FITC, GFP, Alexa 488, CFSE, BB515 |
585/40 | PE,PKH26 | |
670/14 | 7AAD, PE-Cy5, PerCP | |
712/21 | PerCP-Cy5.5, PerCP-eFluor710 | |
780/60 | PE-Cy7 | |
Red (633nm) | 670/30 | APC, Alexa 647 |
780/60 | APC-Cy7, APC-H7, APC-Alexa 750 |
Details
- Laser Excitations: Violet (407nm), Blue (488nm), Red (640nm)
- Colours: 15
- Functions: Analyzer
Configurations
Lasers | Filters | Common Fluorochromes and Dyes |
Violet (407nm) | 450/50 | BV 421, Pacific Blue, Cascade Blue, Alexa 405, V450, Fixable Violet, DAPI |
525/50 | BV510, Alexa 430, V500, AmCyan, Aqua | |
610/20 | BV605, eFluor 605 | |
660/20 | BV650, eFluor 650 | |
712/21 | BV 711 | |
800/50 | BV785 | |
Blue (488nm) | 530/30 | FITC, GFP, Alexa 488, CFSE, BB515 |
575/26 | PE, PKH 26 | |
616/23 | PE-CF594, PE-Texa Red, PI | |
670/14 | PerCP, 7AAD, PE-Cy5 | |
712/21 | PerCP-Cy5.5, PE-Alexa700, PE-Cy5.5, PerCP-eFluor 710 | |
780/60 | PE-Cy7, PE-Alexa 750 | |
Red (640nm) | 670/30 | APC, Alexa 647, Cy5 |
730/45 | Alexa 700, APC-R700, APC-Cy5.5 | |
780/60 | APC-Cy7, APC-Alexa 750, APC -eFluor 780, Live/Dead Fixable Near-IR Dead Cell stain Kit |
Details
- The five-laser Cytek Aurora is a spectral cytometer that measures fluorescence in 64-channels of detection across five avalanche-photodiodes (APD) detector arrays. APDs offer greater sensitivity for fluorochromes which fluoresce at longer wavelengths e.g. PE-Cy7;
- Spectral flow cytometry uses dispersive optics, such as prisms or gratings, to disperse the collected light across a detector array, allowing the full spectra from each particle to be measured;
- The Aurora is equipped with a UV (355nm, 20mW), violet (405nm, 100mW), blue (488nm, 50mW), 50mW yellow-green & red (638nm, 80mW) lasers. The lasers have a flat-top beam profile with a narrow vertical beam height, optimised for the detection of small particles;
- In contrast to traditional flow cytometers which use optical filters to deconvolute fluorescent signals, the Aurora measures the ‘spectral fingerprint’ of different fluorochromes and is able to ‘unmix’ the signals from a multi-stained sample i.e. two fluorochromes that could not be used together using a traditional cytometer can often be identified as separate signals by the Aurora;
- The Cytek Aurora is equipped with a Cytek Automated Sample Loader (ASL), which allows for multiple sample input formats, from plates (96-well) to tubes (48 5mL tubes). A volumetric sensor enables the calculation of counts per microL;
- The Aurora also is able to extract cellular autofluorescence, greatly aiding the analysis of cellular markers in highly autofluorescent cell types or after techniques that increase autofluorescence such as intracellular cytokine staining – something not possible with traditional flow cytometers. Autofluorescence extraction can enhance dim signals and accurately distinguish true signal from sample autofluorescence.
MDCL 5058
Instruments in this Room
Details
- Laser Excitations: Violet (405nm), Blue (488nm), Red (643nm)
- Colours: 10 (Flexible configurations are available for specific experiments)
- Functions: Sorter; 4-way sorting into microtubes and 12x75mm tubes
- Two-way sorting into microtubes, 12x75mm, and 15mL tubes; one-way sorting into 50mL tubes and 6-, 12-, 24-, 48-, and 96-well plates
Configurations
Lasers | Filters | Common Fluorochromes and Dyes |
Violet (405nm) | 450/65 | Pacific Blue, AF405, V450, BFP, Vio Blue, CFP, Zombie violet, BV421, AmCyan |
530/40 | Ametrine, Pacific Orange, BV510 | |
Blue (488nm) | 488/10 | Side Scatter |
530/40 | FITC, GFP, Alexa 488, CFSE | |
580/30 | PE | |
613/20 | PE-Texas Red, PE-AF610, PI | |
670/30 | 7AAD, PE-Cy5, PerCP-Cy5.5 | |
740LP | PE-Cy7, PE-Vio770 | |
Red (643nm) | 670/30 | APC, AF647, iRFP670 |
780/40 | Near-IR Live/dead, APC-Vio770, APC-Cy7 | |
720/12 | AF700 |
MDCL 5144
Instruments in this Room
Details
- Laser Excitations: Violet (405nm), Blue (488nm), Yellow Green (561), Red (633nm)
- Colours: 16 (Flexible configurations are available for specific experiments)
- Functions: Sorter; 4-way sorting into microtubes and 12x75mm tubes,
- Two-way sorting into microtubes, 12x75mm, and 15mL tubes;
- One-way sorting (including single cell) into 6-, 12-, 24-, 48-, 96- and 384-well plates
- Index sorting
Configurations
Lasers | Filters | Common Fluorochromes and Dyes |
Violet (405nm) | 450/40 | V450-PB, BV421, Pacific Blue, eFluor450, V450, Cascade Blue |
525/50 | V525-KrC, BV510, AmCyan, V525, Qdot525, FVS510, AF430, KromeOrange | |
610/20 | V610-BV605, BV605, Qdot605, V610, SuperBright600 | |
670/30 | BV650, StarBrifght Violet 670 | |
710/50 | V712-BV711, BV711, Qdot705, V712, SuperBright702 | |
780/60 | V780, BV785, Qdot800, V780-BV785 | |
Blue (488nm) | 488/10 | Side Scatter |
530/30 | FITC, AF488, GFP, ZombieGreen, CFSE, B525, B525-FITC, Fluo-4 | |
710/50 | PerCP, B675, B675-PerCP, PerCP-Cy5.5, B712, BB700, PerCP-efluor710 | |
Yellow Green (561nm) | 585/15 | PE, Y585-PE, Dtomato, DsRed, Y585 |
610/20 | Y610-mCherry, PE-CF594, ECD, PE-Dazzle594, PI, PE-TexasRed, mCherry, Y610 | |
660/10 | PE-Cy5, mPlum, 7AAD, Y660-PC5, Y660 | |
695/40 | PE-Cy5.5, Y710, PE-AF700 | |
780/60 | Y780-PC7, PE-Cy7, Y780 | |
Red (633nm) | 660/20 | APC, Cy5, R670, AF647 |
730/45 | APC-Cy5.5, AF700, APC-R700, R712 | |
780/60 | APC-Cy7, APC-H7, R780-APC-Cy7, APC-AF750, ZombieNIR |
Details
- Laser Excitations: Near UV (375nm), Volet (405nm), Blue (488nm), Yellow Green (561nm), Red (635nm)
- Colours: 19
- Functions: high efficiency and low-noise Fiber Array Photodiode Detectors (FAPDs) for superior resolution and sensitivity with 7 decades tunable dynamic range of detection;
- an additional side scatter (VSSC) from the violet (405 nm) laser allows the detection of nanoparticles (<200 nm);
- robust and simple plate loading for 96-well plates;
- volumetric measurements with continuous flow, so no need for counting beads for particle enumeration;
- intuitive, easy-to-operate CytExpert acquisition/analysis software;
- compensation values are automatically adjusted in real-time if the gain settings are changed using a compensation library (a repository of compensation spillover values of dyes) due to the linear response across the entire range.
Configurations
Lasers | Filters | Common Fluorochromes and Dyes |
NearUV (375nm) | 450/45 | NUV450, DAPI, Filipin, Hoechst33342,Hoechst Blue, Hoechst33258, Hoechst34580 |
585/15 | NUV585, BV570, Qdot585, FVS575V, Pacific Orange | |
675/30 | NUV675, BUV661, HoechstRed, Qdot655 | |
Violet (405nm) | 450/45 | V450-PB, BV421, Pacific Blue, eFluor450, V450, Cascade Blue |
525/40 | V525-KrC, BV510, AmCyan, V525, Qdot525, FVS510, AF430, KromeOrange | |
610/20 | V610-BV605, BV605, Qdot605, V610, SuperBright600 | |
712/25 | V712-BV711, BV711, Qdot705, V712, SuperBright702 | |
780/60 | V780-BV785, BV785, Qdot800, V780 | |
Blue (488nm) | 525/40BP | FITC, AF488, GFP, ZombieGreen, CFSE, B525, B525-FITC, Fluo-4 |
675/13BP | PerCP, B675, B675-PerCP | |
712/25BP | PerCP-Cy5.5, B712, BB700, PerCP-eflior710 | |
Yellow Green (561nm) | 585/15 | PE, Y585-PE, Dtomato, DsRed, Y585 |
610/20 | Y610-mCherry, PE-CF594, ECD, PE-Dazzle594, PI, PE-TexasRed, mCherry, Y610 | |
660/10 | PE-Cy5, mPlum, 7AAD, Y660-PC5, Y660 | |
710/50 | PE-Cy5.5, Y710, PE-AF700 | |
780/60 | Y780-PC7, PE-Cy7, Y780 | |
Red (635nm) | 670/30 | APC, Cy5, R670, AF647 |
712/25 | APC-Cy5.5, AF700, APC-R700, R712 | |
780/60 | APC-Cy7, APC-H7, R780-APC-Cy7, APC-AF750, ZombieNIR |
Details
- Laser Excitations: Violet (405nm), Blue (488nm), Yellow Green (561nm), Red (635nm)
- Colours: 13
- Functions: high efficiency and low-noise Fiber Array Photodiode Detectors (FAPDs) for superior resolution and sensitivity with 7 decades tunable dynamic range of detection
- an additional side scatter (VSSC) from the violet (405 nm) laser allows the detection of nanoparticles (<200 nm)
- robust and simple plate loading for 96-well plates
- volumetric measurements with continuous flow, so no need for counting beads for particle enumeration
- intuitive, easy-to-operate CytExpert acquisition/analysis software
- compensation values are automatically adjusted in real-time if the gain settings are changed using a compensation library (a repository of compensation spillover values of dyes) due to the linear response across the entire range.
Configurations
Lasers | Filters | Common Fluorochromes and Dyes |
Violet (405nm) | 450/45 | V450-PB, BV421, Pacific Blue, eFluor450, V450, Cascade Blue |
525/40 | V525-KrC, BV510, AmCyan, V525, Qdot525, FVS510, AF430, KromeOrange | |
610/20 | V610-BV605, BV605, Qdot605, V610, SuperBright600 | |
710/50 | V712-BV711, BV711, Qdot705, V712, SuperBright702 | |
Blue (488nm) | 525/40 | FITC, AF488, GFP, ZombieGreen, CFSE, B525, B525-FITC, Fluo-4 |
690/50 | PerCP, B675, B675-PerCP, PerCP-Cy5.5, B712, BB700, PerCP-eflior710 | |
Yellow Green (561nm) | 585/15 | PE, Y585-PE, Dtomato, DsRed, Y585 |
610/20 | Y610-mCherry, PE-CF594, ECD, PE-Dazzle594, PI, PE-TexasRed, mCherry, Y610 | |
660/10 | PE-Cy5, mPlum, 7AAD, Y660-PC5, Y660 | |
780/60 | Y780-PC7, PE-Cy7, Y780 | |
Red (640nm) | 660/10 | APC, Cy5, R670, AF647 |
712/25 | APC-Cy5.5, AF700, APC-R700, R712 | |
780/60 | APC-Cy7, APC-H7, R780-APC-Cy7, APC-AF750, ZombieNIR |
Booking & Policies
Expandable List
- 24hr availability for booking
- Once trained, users can book instruments through our online booking system
- Minimum usage charge is 0.5hr
Schedule changes:
- You are billed on the greater of the time you reserve or the time you use on the flow cytometer
- Instrument use time is calculated from the beginning of your scheduled time to your log out time, unless your log-out time is lower than booked time, in which case your booked time would be charged
- We reserve the right to restrict your access to the facility in the event of frequent last-minute cancellations, late arrivals or not showing up for your appointments at all
- For sorting: you are billed the greater of the time you book or the time you use
- Daily sorter set up and clean up time is billed evenly between users per day, usually 0.5 -1hr depending on the number of users per day.
- Scheduling is on a first-come, first-serve basis
- You must sign up for cytometer time on flow cytometry online calendar in advance – link
- Please be punctual if you have booked the cytometer
- If you finish the experiment significantly earlier than planned, please notify next person or shut down the instrument
- Running over one’s scheduled time is permitted ONLY if the next user agrees. If not, you must yield the cytometer to the next scheduled user
- If you see that the instrument is available and decide to work on a cytometer at the last minute, you must still sign up on the online booking calendar
- Priority is always given to individuals who have reserved time on the website, so if you don’t sign up at the last minute and someone else does, you must stop your work and yield the instrument to the individual who has reserved time on the system
- You MUST cancel online booking if you cancel your experiment. Cancellations must be made at least 3 hours prior to the scheduled time if it’s during working week days 9 a.m. – 6 p.m. If the scheduled time is after hours or on weekends or during holidays you can cancel you booking up to the last minute before the booked time
- Should cancellation happen on a weekend or during the holidays notify person by email booked before or after you
- Failure to make a proper cancellation will result in full time charge
- To book and access for samples to be run with the facility assistance, you must first contact the operator via email or in person
- You must describe your experiment, cell type, fluorochromes, etc.
- The Operator must be given your name and phone number or email, as well as a research billing account number and department of affiliation
- If you must cancel the experiment, please notify the operator at least 24 hours prior to the scheduled time.
- Failure to make a proper cancellation might result in full time charge
- Sorters can only be operated by a designated facility operator
- To sign up, you must first contact designated facility operator by email or phone
- It is required for researchers who need the cell sorting service to go over the sorting experimental design with the operator, please have the following information regarding your sort ready:
-
- Cell type and cell origin
- Number of cells per sample, number of samples to sort
- Types of collection needed, i.e. how many populations you wish to recover
- % of positive cells need to be sorted (if known)
- Fluorochromes being used
- Downstream application for the sorted cells
- Sterility requirements
- Biohazards (Must inform sort operator in advance of potential biohazardous material)
- Account info
- If you cancel the experiment, please notify the operator at least 24 hours prior to the scheduled time via email
- Failure to make a proper cancellation might result in full time charge
You will still be charged for the set-up, cleaning and shutdown, but you are at liberty to cancel the sort at this point, so as not to incur any more unnecessary charges.
The Flow Cytometry and Cell Sorting Core Facility is a dual-level biosafety lab which utilizes Biosafety Level 1 (BSL1) as the minimal work practices level in the primary lab space and Biosafety Level 2 (BSL2) in the facility’s biocabinet.
- You must finish or update the safety training required by the safety office of University annually
- You MUST wear lab coat and gloves at all times and remove them before leaving the facility
- Hands must be washed after handling biohazardous materials and before leaving the facility
- All procedures must be performed carefully to minimize the creation of aerosols
- No unfixed BSL2 samples are allowed to be run on Fortessa and LSR II
- You must prepare samples and dispose of all biohazardous material/waste in your own lab
- Work surfaces are to be decontaminated with 10% freshly-made bleach after any spillage of sample and hazardous material
- For all BSL2 sorting, you must inform the facility operator on the first meeting
- You must follow TDG regulations when you transport unfixed BSL2 samples to and from the facility
- No mouth pipetting, no eating, drinking, or applying cosmetics are allowed in the work area
- No sharps and radioactive material are allowed in the facility
Contact Us
MDCL 4098
MDCL 5058
MDCL 5144
Information Box Group
Minomi Subapanditha
McMaster Immunology Research Centre
Minomi Subapanditha
McMaster Immunology Research Centre
Minomi Subapanditha
McMaster Immunology Research Centre
Minomi Subapanditha
McMaster Immunology Research Centre