Light Microscopy and Raman Imaging of Carotenoids in Plant Cells In Situ and in Released Carotene Crystals
Tomasz Oleszkiewicz , Marta Z. Pacia , Ewa Grzebelus , Rafał Barański
AbstractLight microscopy with a bright field mode offers an easy and fast examination of plant specimen for carotenoid presence in its cells. Using basic techniques such as hand sectioned or squashed preparations, carotenoid-rich chromoplasts can be identified without applying any staining procedure and their localization within the cell, their shape and number can be assessed. More detailed information can be obtained by using Raman spectroscopy which is suitable for the analysis of carotenoids due to their unique Raman spectra and allows semiquantification of their contents. Raman imaging (mapping) can be additionally used to show the distribution of carotenoids within the sample. Raman spectra can be taken from extracted carotenoids but can be also obtained directly from plant tissues or cells as Raman measurements are nondestructive for the sample. Here we describe preparations of intact tissue samples, monolayer cell samples, isolated protoplasts as well as carotene crystals released from chromoplasts that are suitable for subsequent observations using light microscopy and for analysis using Raman spectroscopy.
|Publication size in sheets||0.75|
|Book||Rodriguez-Concepciónn Manuel, Welsch Ralf (eds.): Plant and Food Carotenoids, Methods and Protocols , Methods in Molecular Biology, 2020, Springer, ISBN 978-1-4939-9951-4, [978-1-4939-9952-1], 421 p., DOI:10.1007/978-1-4939-9952-1|
|Keywords in English||Carrot, Chromoplasts, Bright-field microscopy, Nondestructive measurement, Protoplast, Raman mapping, Spectroscopy|
|Finansowanie||The protocol is partially the result of the research project supported by the National Science Centre in Poland (UMO-2013/09/B/NZ9/02379)|
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