Raman Micro-Spectroscopy as a Non-invasive Cell Viability Test

Abstract

The number of techniques to identify, quantify and characterise cell death is rapidly increasing as more is known about the complex mechanisms underlying this process. However, most of these techniques are invasive and require preparation steps such as cell fixation, staining or protein extractions. Non-invasive analysis of living cells represents a key point in cell biology, e.g. in toxicology studies or in tissue engineering. In this chapter, we report the usefulness of Raman spectroscopy as a non-invasive method to distinguish cells at different stages of cell cycle and living cells from dead cells. Throughout two examples, we show the performance and the use of Raman spectroscopy as a new non-invasive method to assess cell viability.

Sub-cellular dynamic investigation of the multi-component drug on the gastric cancer cell BGC823 using Raman spectroscopy

“…The characteristic peaks are assigned in Table (Table S1) based on previous studies involving several cell lines and biomolecules. 31-36 Figure 2 and Figure S3 shows that the Raman spectra profile of treated groups were decreased after 24h, which mainly reflected in those Raman bands assigned to nucleic acid: 794 cm -1 (DNA), 941cm -1 (RNA), 1092 cm -1 (DNA), 1375 cm -1 (A, G, T) 1579 cm -1 (DNA). The changes at 48h shown in figure S1 were similar to those at 24h.…”

Raman spectroscopy and regenerative medicine: a review

“…In an effort to quantify cell viability non-invasively, Notingher et al 67 – 69 used spontaneous Raman spectroscopy to distinguish between live and dead cells and also found that the stage of the cell cycle that particular cells were in could be determined primarily by changes in peaks attributed to DNA at 782, 788 and 1095 cm −1 . Dead cells had particularly low intensity DNA peaks and also alterations in protein peaks, thought to be due to changes in protein conformation.…”

Surface-Enhanced Raman Scattering for Label-Free Living Single Cell Analysis

“…Another class of contributors to the spectral outcome is the instrumental settings. The laser wavelength, numerical aperture of the microscope objective, the use of a high-speed encoded stage (HSES), laser power, laser exposure time, and its focus adjustment are the main instrumental parameters, most of which affect the signal intensity and pattern. − …”