Fast outdoor screening and discrimination of carotenoids of halophilic microorganisms using miniaturized Raman spectrometers
Abstrakt
Eight miniaturized Raman spectrometers were used to perform a fast outdoor screening and discrimination of carotenoids of a series of halophilic and non-halophilic microorganisms on a set of eight lyophilized samples, each containing high concentrations of a specific dominant carotenoid pigment. Raman spectra were acquired using different excitations (532, 785, sequentially shifted excitation of 785 and 853, and 1064 nm), based on the model of each Raman spectrometer, in order to ascertain the feasibility of individual wavelengths.
The wavenumber positions of diagnostic Raman bands of carotenoids were observed for the different carotenoid species. Characteristic carotenoid Raman bands of the pigment bacterioruberin were reported (using the 532 nm excitation) at 1504-1509 cm(-1), salinixanthin at 1510-1513 cm(-1), spirilloxanthin at 1509-1513 cm(-1), decaprenoxanthin at 1519 cm(-1), β-carotene at 1526 cm(-1), and sarcinaxanthin at 1526-1528 cm(-1).
A 532 nm excitation consistently provided best results due to the significant resonance signal enhancement (both quantitative and qualitative carotenoid detection). Good results were also obtained using the sequentially shifted excitation combining two lasers in the near infrared spectral region, and similarly good results were acquired using a standard 1064 nm excitation.
The least suitable was a 785 nm excitation, with the carotenoid Raman signal almost always weaker compared to major fluorescence signal arising from other types of pigments or biomolecules in the samples. A thorough light shielding was essential in order to acquire good quality data.
This study shows that miniaturized Raman spectrometers, some even equipped with longer wavelength excitation, are able to detect different carotenoid pigments under non-laboratory conditions in a fast way, and discriminate between them, to a certain degree. The implications of this type of research are especially useful in astrobiology, where the searching, detection and discrimination of biomarkers such as carotenoids is receiving significant attention.