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Raman Microscopy

The AlgaeScienceCenter developed in an international collaboration an original approach to investigate composition of individual algal cells by confocal Raman microscope. This approach resolved the major obstacle of applying Raman spectroscopy to photosynthetic cells by quenching the strong pigment autofluorescence (Moudříková et al. 2016). The major innovative aspect of the new method is in in-situ quantification of poly-phosphates and in recent discovery of guanine crystals in algal cells.

The LabRam Evolution microscope (Horiba) that was used for this research is equipped with 532 and 785 nm lasers and inverted microscope Eclipse Ti-U (Nikon) and thermoelectrically-cooled, open-electrode CCD detector (Symphony, Horiba Scientific). Due to the combination of the LabRam spectrograph's 800 mm focal length with a grating of 150 grooves mm−1, spectral ranges as wide as 5640 cm−1 and 2580 cm−1 are covered with 532 nm and 785 nm excitation, respectively. The 785 nm laser is used to identify by Raman spectroscopy molecular vibrations of carotenoids while the 532 laser serves to identify and locate in algal cells lipids, proteins, starch, and polyphosphates.

Reference

Š. Moudříková, P. Mojzeš, C. Pfaff, D. Behrendt, L. Nedbal, et al., Raman and fluorescence microscopy sensing energy-transducing and energy-storing structures in microalgae. Algal Research 16: 224-232 (2016).


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