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A dual compartment cuvette system for correcting scattering in whole-cell absorbance spectroscopy of photosynthetic microorganisms.
Hervey, John R D; Bombelli, Paolo; Lea-Smith, David J; Hulme, Alan K; Hulme, Nathan R; Rullay, Atvinder K; Keighley, Robert; Howe, Christopher J.
Afiliação
  • Hervey JRD; Department of Biochemistry, University of Cambridge, Hopkins Building, Downing Site, Tennis Court Road, Cambridge, CB2 1QW, UK.
  • Bombelli P; Department of Biochemistry, University of Cambridge, Hopkins Building, Downing Site, Tennis Court Road, Cambridge, CB2 1QW, UK.
  • Lea-Smith DJ; Department of Biochemistry, University of Cambridge, Hopkins Building, Downing Site, Tennis Court Road, Cambridge, CB2 1QW, UK.
  • Hulme AK; School of Biological Sciences, University of East Anglia, Norwich Research Park, Norwich, NR4 7TJ, UK.
  • Hulme NR; Starna Scientific Ltd, Hainault Business Park, 52/54 Fowler Rd, Ilford, IG6 3UT, UK.
  • Rullay AK; Starna Scientific Ltd, Hainault Business Park, 52/54 Fowler Rd, Ilford, IG6 3UT, UK.
  • Keighley R; Shimadzu UK Limited, Unit 1, Mill Crt, Featherstone, MK12 5RD, UK.
  • Howe CJ; Shimadzu UK Limited, Unit 1, Mill Crt, Featherstone, MK12 5RD, UK.
Photosynth Res ; 151(1): 61-69, 2022 Jan.
Article em En | MEDLINE | ID: mdl-34390453
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cianobactérias / Fótons Idioma: En Revista: Photosynth Res Assunto da revista: METABOLISMO Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cianobactérias / Fótons Idioma: En Revista: Photosynth Res Assunto da revista: METABOLISMO Ano de publicação: 2022 Tipo de documento: Article