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SickKids Centre for Advanced Single Cell Analysis (CASCA)

Special Announcements:

 

New Service November 2020: Hyperion Imaging Mass Cytometry (IMC) 

With this innovative technology you can deeply interrogate tissues and tumors at subcellular (1 mm) resolution to define cell phenotypes, cell-cell interactions and spatial proximity within normal and diseased microenvironments.

  • Highly multiplexed immunohistochemistry analysis of tissue sections stained with metal-tagged antibodies
  • Combines a laser ablation module with the high parameter capabilities of mass cytometry (CyTOF)
  • Simultaneous analysis of up to 37 markers in a single scan

To book a consultation or appointment please see "Request Services" tab

For more information about IMC and related regents, please visit Fluidigm  

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September 2: Phase 3 Changes to our Helios Acquisition Service

Phase 3 Hours

  • Helios acquisitions will be scheduled Monday-Friday 9:30am-3:30pm
  • Only 1 client allowed to enter per appointment
  • Clients will not stay longer than necessary

Bookings/Reservations

  • Clients with permission to book direcly on Helio1 or Helios3 calendar please do so
  • All other clients please submit "Helios Fluidics Acquisition Request" (see "Request Services tab", scroll down to "Helios Fluidics Time")
  • For 9:30am appointments, samples can be dropped off from 8:30-9:30 am
  • For later appointments samples should be dropped off up to 1 hr before

Facility Access

  • External clients need to go through COVID-19 screening
  • Elevator occupancy will be limited to 3 people
  • Please avoid high traffic times
  • Allow extra time to enter PGCRL and reach the 18th floor

Enhanced Biosafety procedures

  • Masking is mandatory in all areas of PGCRL
  • Mask will be provided to external clients at the COVID-19 screening station
  • To minimize contact, we will implement a “no-contact” and paperless sample drop-off system
    • Use iLAB to provide all experiment information
    • Leave your samples on drop-off table (in ice box) just inside the Facility

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CASCA provides fee-for-service access to innovative "next generation" mass cytometry and scRNA-Seq technologies for performing high content single cell analysis to advance fundamental discovery projects as well as “Precision Medicine” research initiatives. Our current technology platforms are briefly described below:

BD Rhapsody Single Cell RNA-Sequencing & Protein "Multi-omics" Analysis Service

Download our Rhapsody service options and pricing or answers to Frequently Asked Questions

  • This microwell-based system provides the ability to simultaneously quantify RNA and surface proteins in up to 40,000 single cells in a single experiment
    • Single cells are gently deposited into wells without causing any stress on the cells
    • Each well is then matched with a BD Rhapsody™ bead that captures the mRNA and oligos from BD AbSeq reagents that were used to pre-label cell surface proteins
    • After cell separation the mRNA and BD™ Ab-Seq Oligos captured from each cell are ready for cDNA synthesis and preparation of indexed Illumina libraries
    • A unique barcode on each bead allows each captured molecule to be matched to its respective cell
    • This system provides automated cell counting, viability measurements and cell capture rates to provide quality control information prior to library preparation and sequencing
  • The BD™ Single-Cell Multiplexing Kit provides the ability to combine hematopoietic cells from 2-12 different samples in one experiment to:
    • Reduce batch effects by simultaneously running multiple controls and experimental samples at the same time
    • Substantially reduce the per sample cost
    • Improve detection and elimination of multiplets from the data
  • Use oligo-conjugated antibodies (BD™ AbSeq reagents) to provide protein expression data similar to flow cytometry
    • Cells can be pre-labelled with up to 40 Ab-Seq reagents to provide deep phenotyping of each cell sequenced
  • With BD™ Rhapsody you can choose between whole transcriptome analysis (WTA) and gene-targeted assays:
    • WTA surveys the entire transcriptome at low sensitivity, requiring high read-depths which increase sequencing costs
    • Immune gene-targeted assays use a panel of specific primer sets to amplify~400 immune genes at high sensitivity with lower sequencing costs
    • You can also purchase custom gene-targeted primers to supplement BD gene panels, or work with BD bioinformaticians to design a custom gene panel

 Download this BD Rhapsody Single-Cell Multiomics System Overview or Contact Tina Chen for more information

Mass Cytometry (CyTOF) Services 

  • Time-of-flight (TOF) mass spectrometer-coupled flow cytometers (known as CyTOFs) are used to analyze single cells stained with antibodies (Abs) or other cellular probes labeled with rare earth metals
  • CyTOF can analyze single cells stained with up to up to 50 metal-tagged probes of cell lineage, differentiation and function
  • Thus, CyTOF overcomes limitations of fluorescence flow cytometry to allow unprecedented depth for profiling single cells within heterogeneous populations from mouse models or patients
  • CASCA now operates 3 Helios instruments, which are 3rd generation CyTOFs
  • We provide biomedical researchers with scientific, technical and bioinformatics support to develop CyTOF applications for both fundamental, translational and pre-clinical research projects

     To learn more about our Mass Cytometry services, please contact Tina Chen 

Using iLab to Access CASCA Services

  • First register for an iLab account by clicking HERE
  • Within one business day you will receive a password and further instructions
  • Please note that iLab functionalities work best with Chrome and Firefox
  • Please contact John Moniakis for help with iLab

Use the Request Services tab to:

  • Schedule a free Consultation for a general introduction to our CyTOF or Rhapsody services
  • Request Training to perform surface and intracellular staining of cells with metal-tagged antibodies
  • Obtain quotations for us to perform a pilot experiment so you can see the advantages of using CyTOF for high content single cell analysis 
  • Request Helios appointments to analyze your cells stained with metal-tagged Abs
  • Requests Consultations, Data Analysis, custom metal-tagging of antibodies, consumables purchases and other services
  • Request BD Rhapsody services

Use the Schedule Equipment tab to:

  • Book appointments on the Helios-1 Calendar
  • Note that this option is only available to experienced clients

 

CASCA Biosafety: CASCA operates in a Containment Level 2 (CL2) laboratory and can handle samples designated as RG1 or RG2. No samples or agents designated as RG3 and RG4 can be brought into the lab

  • All Persons must wear gown and gloves (provided by CASCA) to be donned on entry into the lab and discarded on exit.
  • Samples transported into the CASCA must be in sealed primary containers (FACS tubes, screw cap tubes, etc.). In addition, sealed primary containers must be in a sealed secondary leak proof container.

 

Location and Hours of Operation

Hours Open / Hours Staffed Location

9AM- 5PM

 

Lab Room 18-9430
Peter Gilgan Centre for Research and Learning (PGCRL)
The Hospital for Sick Children
686 Bay Street Toronto, ON M5G 0A4

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