CB 399: Advanced Flow Cytometry Fall 2015

Advanced Flow Cytometry

Course Director: Dr. Tim Mitchison

Curriculum Fellow:  Kevin Bonham (kevin_bonham@hms.harvard.edu) and Emily Gleason

Flow cytometry is a technology that can be used for a wide range of both clinical and research-based applications. It offers qualitative and quantitative data, describing cellular phenotypes and functional properties.  Recent advances in hardware, instrumentation and reagents have now made it possible to use higher-order polychromatic assays of complex design that measure up to 27 parameters. These multicolor assays offer great potential to ask new questions, or provide new answers to old questions. This nanocourse will introduce you to the principles of polychromatic experimental design and cover the relevant controls required for troubleshooting protocols or instrumentation, facilitating accurate gating, compensation and providing a biological comparison.

Lecturers:

  • Dr. Jodene Moore, Director of the HMS Systems Biology Flow Cytometry Facility 
  • Dr. Fabien Depis, Postdoctoral Fellow, Benoist Lab

Schedule: 

First Session (open to the Harvard community): Monday, October 19th 12:30- 4 PM
Location: Systems Biology Conference Room - Armenise 506

Second Session (limited to registered students): Wednesday, October 28th 12:30 – 3 PM
Location: Countway Library, Minot Room

First Session Schedule:

-   How To Think About Flow Cytometry Experiments

  • Hypothesis
  • Identification
  • End Point

-   Basic Tenets of Experimental Design in Flow Cytometry

-   Panel Design

-   Flow Cytometry Controls

  • Instrument Calibration/QC
  • Compensation
  • Gating/FMOs
  • Biological

BREAK-15 minutes

-   Flow Cytometry Applications - Fabien Depis, PhD

Assignment: Design a multicolor flow cytometry experiment based on your current field of study (can include sorting). Your proposal should include the following information:

  1. HYPOTHESIS,
  2. RATIONALE,
  3. EXPERIMENTAL DESIGN,
  4. WORKFLOW (Sample Processing Steps), and
  5. CONTROLS.
  6. YOUR GATING STRATEGY
  7. PLOTS with parameter labels demonstrating what your data would look like under conditions where your hypothesis is proven true and proven false.

                                                            OR

If your experimentation just takes you up to a sorting step, show those plot(s).

You may add a final experimental technique(s) (i.e. RNA seq, PCR etc.) section. Student completed the Introductory Flow Cytometry Nanocourse may reuse their hypothesis and rationale for this assignment.

Please email your proposals to Kevin Bonham (kevin_bonham@hms.harvard.edu)by 5pm Sunday, October 25th. Kevin will compile all of the assignments and share them with the class. Please come to the second session prepared to discuss your proposal and those of your classmates.

DROP DEADLINE: Monday, October 12, 2015

 AUDITORS (Post-Docs, Faculty, or Staff) DO NOT NEED TO SIGN UP TO ATTEND THE 1st SESSION.  PLEASE DO NOT ENROLL