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Gamma-proteobacteria in seawater

Gamma-proteobacteria in seawater. Sidhartha Goyal Hyunsoo Na. Contents. Introduction on (CARD-)FISH Procedures Results Discussion. Fluorescent in-situ hybridization. Detection and localization - U nculturable bacteria - Specific mRNAs in tissue

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Gamma-proteobacteria in seawater

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  1. Gamma-proteobacteria in seawater Sidhartha Goyal Hyunsoo Na

  2. Contents • Introduction on (CARD-)FISH • Procedures • Results • Discussion

  3. Fluorescent in-situ hybridization • Detection and localization- Unculturable bacteria- Specific mRNAs in tissue • Capture live bacteria by targeting rRNA

  4. CARD-FISH Cy3 DAPI

  5. Procedures Sample preparation - Fixation and immobilization Permeabilization & inactivation of endogenous peroxidases Hybridization CARD Microscopy

  6. Results – relative abundance • Requires training! • Can Beach sample

  7. Results – cells/ ml at C1/M1 - 200m • Requires training! • Can

  8. Results – cells/ml at Hopkins beach • Requires training! • Can

  9. Distribution of cells on the filter Cy3 DAPI

  10. Discussion • Training - Distinguish between cells and particles • Probe design may be challenging • Consortium analysis • Physical association of cells • Community composition…? • Cheap and quick! `

  11. Thank you

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