Abstract
A multichannel kinetic spectrophotometer–fluorimeter with pulsed measuring beam and differential optics has been constructed for measurements of light-induced absorbance and fluorescence yield changes in isolated chlorophyll-proteins, thylakoids and intact cells including algae and photosynthetic bacteria. The measuring beam, provided by a short (2 µs) pulse from a xenon flash lamp, is divided into a sample and reference channel by a broad band beam splitter. The spectrum in each channel is analyzed separately by a photodiode array. The use of flash measuring beam and differential detection yields high signal-to-noise ratio (noise level of 2 × 10−4 in absorbance units per single flash) with negligible actinic effect. The instrument covers a spectral range between 300 and 1050 nm with a spectral resolution of 2.1, 6.4 or 12.8 nm dependent on the type of grating used. The optical design of the instrument enables measuring of the difference spectra during an actinic irradiation of samples with continuous light and/or saturation flashes. The time resolution of the spectrophotometer is limited by the length of Xe flash lamp pulses to 2 µs.
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Abbreviations
- CCD:
-
Charge coupled device
- Chl:
-
Chlorophyll
- D1:
-
psbA gene product
- LED:
-
Light emitting diode
- PDA:
-
Photodiode array
- Pheo:
-
Pheophytin
- PS II:
-
Photosystem II
- RC:
-
Reaction centre
- T–S:
-
Triplet-minus-singlet
- UV–VIS–NIR:
-
Ultraviolet–visible–near infrared
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Acknowledgements
Authors would like to thank Dr. L. Nedbal (Institute of Systems Biology and Ecology, Academy of Sciences, Czech Republic) for fruitfull discussions, Ing. M. Trtílek (Photon Systems Instruments, Czech Republic) for modifying and assembling the electronics, J. Vítek (Optovid, Czech Republic), Z. Melich (Optical Development Workshop, Academy of Sciences, Czech Republic) and M. Novotný (Technical school, Czech republic) for manufacturing fibre optics, special optical components and metal components, respectively, and Juan B Arellano (Inst. Natural Resources and Agrobiology, Salamanca, Spain) for providing the bacterial reaction centre samples. This work was supported by grants MSMT VS96085, LN00A141, MSM6007665808, AV0Z50510513, GACR 206/97/0325, 206/00/1051, 206/03/1107 and 521/03/H160.
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Bína, D., Litvín, R., Vácha, F. et al. New multichannel kinetic spectrophotometer–fluorimeter with pulsed measuring beam for photosynthesis research. Photosynth Res 88, 351–356 (2006). https://doi.org/10.1007/s11120-006-9071-y
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DOI: https://doi.org/10.1007/s11120-006-9071-y