Raman spectroscopy and imaging of β-carotene in live corpus luteum cells

• Published in: Animal reproduction science 2002-06, Vol.71 (3), p.249-266
• Main Authors: Arikan, Ş., Sands, H.S., Rodway, R.G., Batchelder, D.N.
• Format: Article
• Language: English
• Subjects: Animals; beta Carotene - analysis; beta Carotene - pharmacology; Carotene; Cattle; Cattle-ovary; Cells, Cultured; Chromatography, High Pressure Liquid; Corpus Luteum - chemistry; Corpus Luteum - ultrastructure; Female; Luteal cells; Microsomes - chemistry; Raman spectroscopy; Spectrum Analysis, Raman
• ISSN: 0378-4320; 1873-2232
• DOI: 10.1016/S0378-4320(02)00020-9

Raman spectroscopy has been used to identify and locate β-carotene within individual living luteal cells. The cells were either freshly prepared or cultured; the latter was incubated in the presence or absence of β-carotene in the form of enriched bovine high-density lipoprotein. Luteal cells were investigated using several Raman spectroscopic and imaging techniques. These techniques did not give accurate concentration levels of β-carotene within parts of the cell but illustrated the distribution of the molecule. Freshly prepared luteal cells were found to contain an appreciable concentration of β-carotene. Over a period of several days, the concentration gradually reduced to a nearly undetectable level; similar results were found for cells cultured in the absence of the β-carotene. For cells cultured in the presence of β-carotene, the molecular concentration was maintained for as long as 2 weeks. The Raman spectra of fragmented cells showed that the β-carotene is predominantly localised in the lipid-rich cell components, with the concentration highest in the microsomal fraction. The Raman imaging techniques revealed that β-carotene was spread over the entire volume of the luteal cells with higher levels occurring at distinct sites, including the surface.