Abstract

Label-free Raman Imaging of Living Mammalian Cells - A Valuable New Tool for Investigating Complex Cellular Systems

Katherine Hollywood, Research Associate, Manchester University

Herein we will discuss the applicability of Raman microspectroscopy as a powerful label-free technique for the analysis of single mammalian cells and the adaptability of the technique to undertake live cell Raman mapping, via the incorporation of an on-line incubator within the microscope chamber that allows one to control temperature, humidity and air. Raman mapping offers a powerful opportunity for the analysis of single cells due to its attributed high spatial resolution (< 1 µm), speed of data acquisition and non-invasive and non destructive nature. Additionally, as water is a weak Raman scatterer, spectral contributions pertaining from aqueous cell content or cell culture medium is minimal. Raman mapping is capable of providing vast amounts of spatially resolved biochemical information regarding the composition of cellular and sub-cellular components. A wealth of biological information is detailed within a single Raman spectrum including, amongst other things proteins (amino acids and conformational structures), nucleic acids (DNA and RNA) as well as lipids (cholesterol and CH2 groups). We will present a selection of our current work that will demonstrate the applicability and diversity of this analytical approach. This will include our results from a study in which we have monitored the uptake and localization of a small molecule drug within living keratinocyte cells. Further results will include those collected during the monitoring of induced cell differentiation in a model pancreatic cell line.