Post on 19-Jan-2016
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LAB FOUR Photosynthesis
4A Plant Pigment Chromatography
4B Tracking the Light Reaction
LAB FOUR Photosynthesis
•How is the light reaction affected by the presence or absence of light?
•How is it affected by the boiling of chloroplasts?
LAB FOUR Photosynthesis
Isolated chloroplasts
LAB FOUR Photosynthesis
Isolated chloroplasts
LAB FOUR Photosynthesis
boiled unboiled
LAB FOUR Photosynthesis
Isolated chloroplasts
boiled unboiled
LAB FOUR Photosynthesis
Isolated chloroplasts
boiled unboiled
LAB FOUR Photosynthesis
Isolated chloroplasts
light
dark
LAB FOUR Photosynthesis
Isolated chloroplasts
DPIP
LAB FOUR Photosynthesis
Isolated chloroplasts
DPIP
LAB FOUR Photosynthesis
Isolated chloroplasts
boiled unboiled boiled unboiled
LAB FOUR Photosynthesis
Isolated chloroplasts
boiled unboiled boiled unboiled blank
LAB FOUR Photosynthesis
Isolated chloroplasts
DPIP replaces NADP+ in the light reactions of photosynthesis.
LAB FOUR Photosynthesis
Isolated chloroplasts
DPIP replaces NADP+ in the light reactions of photosynthesis.
As DPIP is reduced, it turns from blue to colorless.
LAB FOUR Photosynthesis
Isolated chloroplasts
DPIP replaces NADP+ in the light reactions of photosynthesis.
As DPIP is reduced, it turns from blue to colorless.
A spectrophotometer measures the absorption of blue light.
0%
LAB FOUR Photosynthesis
Isolated chloroplasts
DPIP replaces NADP+ in the light reactions of photosynthesis.
As DPIP is reduced, it turns from blue to colorless.
A spectrophotometer measures the absorption of blue light.
10%
LAB FOUR Photosynthesis
Isolated chloroplasts
DPIP replaces NADP+ in the light reactions of photosynthesis.
As DPIP is reduced, it turns from blue to colorless.
A spectrophotometer measures the absorption of blue light.
40%
LAB FOUR Photosynthesis
Isolated chloroplasts
DPIP replaces NADP+ in the light reactions of photosynthesis.
As DPIP is reduced, it turns from blue to colorless.
A spectrophotometer measures the absorption of blue light.
80%
Cuvettes:
1.Blank (H2O only)
2.In the dark
3.Normal
4.Boiled chloroplasts
5.No chloroplasts
LAB FOUR Photosynthesis
LAB FOUR Photosynthesis
1 2 3 4 5Light Yes No Yes Yes Yes
Buffer 1 mL 1 mL 1 mL 1 mL 1 mL
H2O 4 mL 3 mL 3 mL 3 mL 3 mL + 3 drops
DPIP 1 mL 1 mL 1 mL 1 mL 1 mL
Chloro-plasts
3 drops 3 drops 3 drops 3 drops boiled
No
Cuvettes:
Keep all cuvettes on ice!
Adjust empty spectrophotometer to 0%
Adjust with blank cuvette to 100%
Add 3 drops chloroplast to your cuvette and immediately measure transmittance.
Measure again at 5, 10, and 15 minutes.
LAB FOUR Photosynthesis
•Five teams will handle one cuvette each•Each team will take four measurements•Your turn lasts exactly one minute.
LAB FOUR Photosynthesis
LAB FOUR Photosynthesis
1 2 3 4 5Light Yes No Yes Yes Yes
Buffer 1 mL 1 mL 1 mL 1 mL 1 mL
H2O 4 mL 3 mL 3 mL 3 mL 3 mL + 3 drops
DPIP 1 mL 1 mL 1 mL 1 mL 1 mL
Chloro-plasts
3 drops 3 drops 3 drops 3 drops boiled
No
Cuvettes:
Keep all cuvettes on ice! .
Adjust empty spectrophotometer to 0%
Adjust with blank cuvette to 100%
Add 3 drops chloroplast to your cuvette and immediately measure transmittance.
Measure again at 5, 10, and 15 minutes.
.
LAB FOUR Photosynthesis