In vivo- and in vitro-analysis of single diatom cells/ cell walls by optical imaging using Raman spectroscopy and non-linear microscopy

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Raman spectroscopy is an important analytical tool which can be applied in order to non-destructively obtain molecular information from integer cells. Based on our ongoing collaboration concerning the study of single diatom cell walls, Raman spectroscopic methods will be developed and used within the proposed project in order to study the cell wall composition as well as the metabolic processes taking place during the formation of the micro- and nanostructured diatom cell walls. The measurements include the study of single, extracted cell walls as well as the study of living cells by the combination of micromanipulation systems with imaging techniques. Non-linear microscopy based on second harmonic generation, two-photon fluorescence and coherent anti-Stokes Raman scattering will be applied additionally. The aim of the project is the investigation of variations of the nature and composition of organic molecules found during the cell cycle. Furthermore, the response of the cells upon defined variations of the composition of the growth medium will be monitored. Optical traps allow Raman experiments on living single diatom cells. In vivo online monitoring will be carried out by combining optical traps and Raman spectroscopy in a microfluidics environment. Furthermore, Raman microscopy will also be performed on extracted single cell walls in combination with other analytical techniques (ICP-OES, NMR) especially with respect to the inorganic components found in the cell walls. Chemometrics will be important for the interpretation of the data.