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Photosynthesis Lab Manual (BIOL 4160)

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In these lab exercises, you will be introduced to a variety of techniques that are very useful in photosynthesis (and other) research. The exercises are designed to allow you to improve your 'Experimental Hands' as you progress from simpler to more complex protocols.

Origins and Fate of Light: Students are introduced to the nature of light and its interactions with photosynthetic structures.

Lab 01 Spectral Properties of Intact Leaves: Students will measure the reflectance and transmittance spectra of intact photosynthetic leaves.

Lab 02 Visualizing the Photosynthetic Apparatus: Students will examine the chloroplast in situ and in vitro, and be introduced to the technique of chloroplast isolation from photosynthetic tissues.

Lab 03 Absorbing Light: Students will collect absorption spectra from chloroplasts in vitro, and from pigments isolated from the chloroplasts.

Fluorescence and Reaction Centers: Students are introduced to fluorescence spectroscopy techniques.

Lab 04 Determining Excitation and Emission Spectra: Students measure the excitation/emission spectra of cells and isolated chloroplasts, and compare them with the excitation/emission of isolated pigments.

Lab 05 Physiology of Fluorescence: Students measure how modifying the physiology of photosynthesis affects fluorescence emission, a metric of the photochemical mechanisms of reaction centers.

Oxygen Electrode: Pathways of Photosynthetic Electron Transport: Students are introduced to the use of the oxygen electrode to assess electron transport chain mechanisms in photosynthesis.


Lab 06 P/O Ratios: Students measure the effect of ADP addition on oxygen production of isolated chloroplasts, to determine the stoichiometry of ATP synthesis to oxygen evolved.

Lab 07 Carbon Dioxide Coupling in Chloroplasts: Students measure the effect of providing carbon dioxide on oxygen production of isolated chloroplasts, to determine whether ATP and NADPH utilization in the C3 pathway increases the rate of NADPH and ATP supply (and thus oxygen production) in intact chloroplasts.

Chloroplast Molecular Biology: Students are introduced to chloroplast genome(s).

Lab 08 Isolation of Chloroplast DNA: Students isolate the chloroplast genomic DNA.

Lab 09 Chloroplast Genome Mapping: Students map the chloroplast genome using restriction endonucleases.

Lab 10 Presentations: Pathways of Carbon Dioxide Fixation: Students give a presentation on an enzyme from one of the carbon dioxide fixation pathways.

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(2011--archived) Lab 01 Chloroplasts in Photosynthetic Tissues: Students are introduced to the technique of chloroplast isolation and compare in situ and in vitro chloroplasts with phase and fluorescence microscopy to demonstrate successful isolation and explore the concept of intact (double membrane bound) and non-intact (naked thylakoids) chloroplast structures.

(2011--archived) ATP Production: Students make direct measurements of ATP production in chloroplasts using the luciferin/luciferase assay.

(2011--archived) Lab 06 Light-Induced ATP Production in Isolated Chloroplasts: Students measure ATP production in illuminated chloroplasts.

(2011--archived) Lab 07 Acid-Base Induced ATP Production in Isolated Chloroplasts: Students measure ATP production by pH gradients: a foundational experiment proving the Chemi-Osmotic Hypothesis of bioenergetics.

  • Created and maintained by RR Lew, Biology Department at York University (with assistance from Miranda Lorenti and Debbie Freele)
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