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Encyclopedia of Experiments

Bacterial Cell Staining and Preparation for Immunological Studies

This video demonstrates the staining and preparation of bacterial cells for immunological studies using red fluorescent dye. The stained bacteria are diluted and resuspended in a growth medium to achieve the desired bacterial concentration for host cell infection, representing the multiplicity of infection.

1. Bacterial growth and staining

  1. Perform all bacterial work in a Biosafety Cabinet, Biosafety Level 2 laboratory.
  2. Inoculate all bacterial cultures, including Pseudomonas aeruginosa (PAO1), Escherichia coliListeria monocytogenes,  Bacillus subtilis, etc., from the frozen glycerol stock and grow them in Tryptic Soy Broth (TSB, 3 mL) in a shaking incubator at 37 °C overnight maintained at 250 rpm.
  3. T.......

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