Flow Cytometry

The McMaster Core Flow facility provides access to operator-assisted sorters and to several state-of-the-art analytical flow cytometers, as well as comprehensive training and consultation services to enable our users to use this technology to elevate the scope and quality of their research. For immunological analysis specifically of materials obtained from human clinical trials; please visit the Human Immune Testing Suite(HITS) website. http://hitsimmune.com/

Flow facility Sites

  • MDCL 4098

Available instruments:

Analysers: BD LSRFortessa, BD LSRII

Sorter: AriaIII

Facility Operator: Hong Liang, This email address is being protected from spambots. You need JavaScript enabled to view it.

Ext: 21589

  • MDCL 5144

Available instruments:

Analyzers: BD LSRII, Miltenyi MacsQuant,

Sorter: BD AriaIIu

Facility Operator: Zoya Tabunshchyk, This email address is being protected from spambots. You need JavaScript enabled to view it.

Ext: 26717

  • MDCL 5058

Available instruments:

Sorter and operator-assisted analyzer: Beckman Coulter MoFlo XDP

Facility Operator: Minomi Subapanditha, This email address is being protected from spambots. You need JavaScript enabled to view it.

This email address is being protected from spambots. You need JavaScript enabled to view it.">Ext: 26382

 

Hours of Operation

Analytical flow cytometers (BD Fortessa, BD LSRII and Miltenyi MAcsQuant) are available to trained users (with passwords) 24 hours/day – 7 days/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.

Fees

Effective as of February 1st, 2019

Analysers

Unassisted/hr

Assisted/hr

 

Sliding scale rate*

BD Fortessa (MDCL4089)

sliding scale

80

 

1 - 120 hrs - $70/hr

BD LSRII (MDCL4089)

sliding scale

80

 

121 - 240 hrs - $50/hr

BD LSRII (MDCL5144)

sliding scale

80

 

After 240hrs - $30/hr

Miltenyi MacsQuant (MDCL5144)

50

80

 

*credits generated starting May 1, 2019

MoFlow (MDSL5058)

N/A

80

   
         
         

Sorters

Unassisted/hr

Assisted/hr

   

AriaIII (MDCL4089)

N/A

80

   

AriaIIu (MDCL5144)

N/A

80

   

MoFlow (MDSL5058)

N/A

80

   
         

Training

80/hr

     

 

 

Instrumentation per site: description, lasers, channels

  

Instrument: BD AriaIII

Room location: MDCL 4089

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,

2-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.

 

Instrument: BD LSRFortessa

Room location: MDCL 4089

Laser Excitations: Violet (407nm), Blue (488nm), Red (640nm)

Colours: 15

Functions: Analyzer

 

Instrument: BD LSRII

Room location: MDCL 4089

Laser Excitations: Violet (405nm), Blue (488nm), Red (633nm)

Colours: 15

Functions: Analyzer

 

Instrument: BD AriaIIu

Room location: MDCL 5144

Laser Excitations: Violet (407nm), Blue (488nm), Red (633nm)

Colours: 9 (Flexible configurations are available for specific experiments)

Functions: Sorter; 4-way sorting into microtubes and 12x75mm tubes,

2-way sorting into microtubes, 12x75mm, and 15mL tubes;

one-way sorting (including single cell) into 6-, 12-, 24-, 48-, 96- and 384-well plates.

 

Instrument: BD LSRII

Room location: MDCL 5144

Laser Excitations: Violet (404nm), Blue (488nm), Yellow Green (561nm), Red (640nm)

Colours: 10

Functions: Analyzer

Instrument: Miltenyi MacsQuant

Room location: MDCL 5144

Laser Excitations: Violet (405nm), Blue (488nm), Red (635nm)

Colours: 8

Functions: Analyzer, equipped with a MiniSampler for automated 96-well plate reading

  

Instrument: MoFlo XDP

Room location: MDCL 5058 (inside MDCL 5059)

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,

2-way sorting into microtubes, 12x75mm, and 15mL tubes; one-way sorting into 50mL tubes and 6-, 12-, 24-, 48-, and 96-well plates.

 

MDCL 4089

Instruments   Lasers          Filters            Common Fluorochromes and Dyes

BD Aria III (Default)

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

       

BD LSRII

Violet (405nm)

450/50

BV 421, Pacific Blue, Cascade Blue, Alexa Fluor 405, V450, Fixable Violet, DAPI

   

525/50

BV510, Alexa 430, V500, AmCyan, Aqua

   

585/15

BV570, Pacilfic Orange, Yellow live/dead fixable cell stain

   

605/12

BV605, eFluor 605

   

655/8

BV650, eFluor 650

   

800/40

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-Alexa 700, PE-Cy5.5, PerCP-eFluor 710

   

780/60

PE-Cy7, PE-Alexa 750

 

Red (633nm)

660/20

APC, Alexa 647, Cy5

   

730/45

Alexa 700, APC-R700, APC-Cy5.5

   

780/60

APC-Cy7, APC-H7, Live/Dead Fixable Near-IR Dead Cell stain Kit

       

Fortessa

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

       
       

 

MDCL 5144

 

Instruments

 

Lasers                

 

Filters          

Common Fluorochromes and Dyes

BD AriaIIu

Violet (407nm)  

450/40

Pacific Blue, Cascade Blue, Alexa 405, V450, Fixable Violet

(Default)

 

605/12

BV605, eFluor 605

 

Blue (488nm)

488/10

Side Scatter

   

530/30

FITC, GFP, Alexa 488, CFSE

   

585/42

PE, DsRed

   

616/23

PE-Texas Red

   

675/20

7AAD, PE-Cy5, PerCP

   

780/60

PE-Cy7

 

Red (633nm)

660/20

APC

   

780/60

APC-Cy7

       

BD LSRII

Violet (404nm)

450/50

Pacific Blue, Cascade Blue, Alexa 405, V450, Fixable Violet

   

605/12

BV605, eFluor 605

 

Blue (488nm)

488/10

Side Scatter

   

530/30

FITC, GFP, Alexa 488, CFSE

   

695/40

7AAD, PerCP-Cy5.5

 

Yellow Green (561nm)

582/15

PE, dsRed

   

610/20

PE-Texas Red, mRFP

   

780/60

PE-Cy7

 

Red (640nm)

670/30

APC, Alexa 647

   

730/45

Alexa 700

   

780/60

APC-Cy7

       

Miltenyi MacsQuant

Violet (405nm)    

450/50

Pacific Blue, Cascade Blue, Alexa 405, V450, Fixable Violet

   

525/50

Alexa 430, AmCyan, Pacific Orange, Cascade Yellow,

 

Blue (488nm)  

488/10

Side Scatter

   

525/50

FITC, GFP, Alexa 488, CFSE

   

585/40

PE, Cy3

   

655-730

7AAD, PE-Cy5, PE-Cy5.5, PerCP-Cy5.5, PI

   

750 LP

PE-Alexa 750, PE-Cy7

 

Red (635nm)

655-730

APC, Alexa 647, Alexa 700, DRAQ5, Cy5

   

750 LP

APC-Cy7, APC-Alexa 750

 

 

 

MDCL 5058

 

Instruments

 

Lasers                

 

Filters          

Common Fluorochromes and Dyes

MoFlo XDP

(Default)

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

 

 Booking and Policies

24hr availability for booking.

Once trained, users can book instruments through our online booking system -> https://booked.mcmaster.ca/flow.

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 -1 hr depending on the number of users per day.

Scheduling and Cancellation for Flow analyzer, self-users:

  • 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 9am – 6pm. 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.

Scheduling and Cancellation for sorters (AriaIII, AriaIIu and MoFlo)

  • 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:
  1. Cell type and cell origin.
  2. Number of cells per sample, number of samples to sort.
  3. Types of collection needed, i.e. how many populations you wish to recover.
  4. % of positive cells need to be sorted (if known)
  5. Fluorochromes being used.
  6. Downstream application for the sorted cells.
  7. Sterility requirements.
  8. Biohazards (Must inform sort operator in advance of potential biohazardous material).  
  9. 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. 

Scheduling and Cancellation for Flow analyzer with the facility assistance      

  • 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.

Safety Rules

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

Hong Liang, This email address is being protected from spambots. You need JavaScript enabled to view it., phone - ext. 21589

Zoya Tabunshchyk, This email address is being protected from spambots. You need JavaScript enabled to view it., phone – ext. 26717

Minomi Subapanditha,This email address is being protected from spambots. You need JavaScript enabled to view it., phone - ext. 26382

 

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