Western New York Flow Users Group

Save the Date

13 July 2011

Registration is open!

Before the Meeting

July 12 - FlowJo Training

Details will be emailed to registered attendees.

After the Meeting

July 14 - Two 1/2 day courses taught by David Galbraith

9am-noon: Fluorescent proteins in flow

1pm-4pm: Non-mammalian cell flow cytometry

Course 1: Fluorescent Proteins in Flow

  • I. Introduction to Fluorescent Proteins
    • What are they,
    • A little history, how they were discovered
    • Why they are so useful.
    • How they are made, and how they are used in experiments.
    • Varieties currently available, and sources.
    • Recommendations of FPs for use in flow.
  • II. Expressing Fluorescent Proteins for use in flow and image cytometry
    • Methods of transformation, transfection, and production of transgenotes.
    • Laser and detection configurations.
    • Compensation.
    • New lasers.
    • Subcellular targeting of FPs.
    • Stability and toxicity issues.
  • III. Practical Examples of Analysis of FP Expression using Flow and Image
    Cytometry
    • Mammalian cells in culture.
    • Plant cells and organs.
    • Complex animal organs.
    • Split FP reconstitution.
    • The “Brainbow”.
    • Kinetic FPs and nano-scale microscopy.
    • Noise in gene expression.

Course 2: Flow Cytometry and Sorting of Organisms other than within the Mammalia

  • I. General Introduction outlining the issues that need to be confronted.
  • II. Problems associated with complex multicellular tissues.
    • Production of single cell suspensions, or
    • Production and analysis of homogenates.
  • III. Dealing with large objects.
    • General introduction to sorting theory.
    • How to use conventional instruments with large particles.
    • Specialized instruments -- the Union Biometrica COPAS and BioSort.
  • IV. Dealing with small objects.
    • Bacteria and smaller life forms, as analyzed using conventional sorters.
    • Biosafety issues.
  • V. Analyzing homogenates and complex mixtures.
    • Plants.
    • Aquatic samples.
  • VI. Case studies: working with model organisms.
    • Yeast.
    • Drosophila
    • Other insects
    • Your favorite organism (general discussion)
  • VII. Case studies: Working with non-model and otherwise unusual organisms.