Respiration, photosynthesis, fluorescence microscopy/staining

3 Pages
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Department
Life Science
Course Code
LIFE 212
Professor
Farida Safadi- Chamberlain

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Description
7 November Results of respiration and photosynthesis Mitochondria/respiration Suspension buffer + MB → -ve Control Pellet (700 x g) + MB → Chloroplasts Supernatant (700 x g) + MB → Mitochondria Chloroplasts/Hill Reaction Chloroplasts: DCPIP acts as a final electron acceptor: gets reduced directly by ETC/quantitative assay Mitochondria: MB gets reduced indirectly by the depletion of oxygen from the media in the presence of active mitochondrial ETC/qualitative assay Fluorescence Microscopy: Concepts Detect, localize, and quantify Proteins, molecules, and structures Uses flurophores stains: fluorescent molecules Can co-stain multiple structures in the same cell Direct or indirect stains Protocol: reasons for steps Types of fluorescence staining Direct staining: fluorescent dye (fluorochrome) directly binds to cellular components Example: Dapi and Hoechst/nucleic acids Indirect Immunostaining: antigen-antibody interaction Flurophore conjugated to a probe that binds to specific structures e.g. Phalloidin Fusion proteins: Green fluorescent protein (GFP) GFP gene is fused to gene of interest
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