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Flow Cytometry at San Jose State University

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Experimental Microbiology

Students will assimilate their knowledge of microbiology, fluorescence microscopy and flow cytometry to answer an experimental question about Tetrahymena nutrition. Tetrahymena will be given prey (fluorescent-labeled yeast or beads) under specific conditions that students will monitor. Students will obtain data using florescence microscopy and a flow cytometer, and perform analysis using Paint-A-GatePROTM software. The question, experimental design, experimentation, and the results analysis are the responsibility of the student. The laboratory instructor will act as a resource for ideas and strategies but is not responsible for experimental results. There are no wrong answers, but results are often ambiguous or inconclusive. Think of the lab section as a team addressing a biological phenomenon. Chances are that the experiment you design has not been performed before, and the results you obtain are unique.

 

The experiment will occur in stages (Worksheets #1, 2 and 3) prior to submitting results to your laboratory instructor. Although you may generate one or a few data sets, you will be responsible for analyzing and summarizing the entire class project. The worksheets are designed to develop increasing knowledge about experimental microbiology and flow cytometry, as detailed below:

Worksheet 1:
  • Microbiology
  • Microscopic observation
  • Cell characterization
Worksheet 2:
  • Flow cytometry
  • Fluid flow system
  • Optics
  • Side scatter
  • Forward scatter
  • Fluorescence
  • Dot plots
  • Paint-A-Gate AcademicTM analysis software
  • Painting
  • Gating
  • Quantifying
Worksheet 3:
  • Scientific investigation
  • Hypothesis development
  • Establishment of an information base
  • Experimental design
  • Experimentation
  • Data acquisition
  • Data analysis

Adapted, in part, with permission from Sally Pasion, Department of Biological Sciences, San Francisco State University.

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