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Hervey, John R. D., Bombelli, Paolo, Lea-Smith, David J. 
ORCID: https://orcid.org/0000-0003-2463-406X, Hulme, Alan K., Hulme, Nathan R., Rullay, Atvinder K., Keighley, Robert and Howe, Christopher J.
(2022)
A dual compartment cuvette system for correcting scattering in whole-cell absorbance spectroscopy of photosynthetic microorganisms.
Photosynthesis Research, 151.
pp. 61-69.
ISSN 0166-8595
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Abstract
Absorption spectroscopy is widely used to determine absorption and transmission spectra of chromophores in solution, in addition to suspensions of particles, including micro-organisms. Light scattering, caused by photons deflected from part or all of the cells or other particles in suspension, results in distortions to the absorption spectra, lost information and poor resolution. A spectrophotometer with an integrating sphere may be used to alleviate this problem. However, these instruments are not universally available in biology laboratories, for reasons such as cost. Here, we describe a novel, rapid, and inexpensive technique that minimises the effect of light scattering when performing whole-cell spectroscopy. This method involves using a custom made dual compartment cuvette containing titanium dioxide in one chamber as a scattering agent. Measurements were conducted of a range of different photosynthetic micro-organisms of varying cell size and morphology, including cyanobacteria, eukaryotic microalgae and a purple non-sulphur bacterium. A concentration of 1 mg ml−1 titanium dioxide, using a spectrophotometer with a slit width of 5 nm, produced spectra for cyanobacteria and microalgae similar (1–4% difference) to those obtained using an integrating sphere. The spectrum > 520 nm was similar to that with an integrating sphere with the purple non-sulphur bacterium. This system produced superior results to those obtained using a recently reported method, the application of the diffusing agent, Scotch™ Magic tape, to the side of the cuvette. The protocol can be completed in an equivalent period of time to standard whole-cell absorbance spectroscopy techniques, and is, in principle, suitable for any dual-beam spectrophotometer.
| Item Type: | Article |
|---|---|
| Additional Information: | Funding Information: John Hervey and David Lea-Smith were funded by the Waste Environmental Education Research Trust. Paolo Bombelli was funded by the Leverhulme Trust (RPG-2015-393) and The Italian Ministry of University and Research (MIUR), within the SIR2014 Grant, project RBSI14JKU3. |
| Uncontrolled Keywords: | absorption spectroscopy,chromophores,cyanobacteria,light scattering,microalgae,whole cell spectra,biochemistry,plant science,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1303 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Science > Research Groups > Molecular Microbiology |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 19 Aug 2021 00:30 |
| Last Modified: | 23 Oct 2022 02:55 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/81148 |
| DOI: | 10.1007/s11120-021-00866-8 |
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