Establishing the extent of sample damage and safe limits of laser exposure during live cell analysis with raman spectroscopy

Student thesis: Doctoral Thesis

Abstract

For a technique to be applied to any system or sample with a need for robust reproducibility, it must undergo rigorous testing and have well defined protocols and procedures. Raman spectroscopy still currently suffers from instrument and user variability, as well as not having enough well defined, set procedures which can be transferred from lab to lab. Raman systems are equipped with a variety of hardware and data processing solutions, but still require better established standard operating procedures, sample interaction and data handling procedures before Raman spectroscopy will be able to be translated as a benchmark technique for chemical analysis and to facilitate the comparison of Raman data between studies and different geographical sites. As Raman spectroscopy is a technique which uses a high-powered light illumination source, there is also a risk of sample damage due to laser exposure; this is particularly concerning for light and temperature sensitive samples such as live cells or other biological tissues. To uphold data integrity, it is important to ensure chemical changes - which can be indicating factors to aid medical diagnosis - are due to the process under study, and not artefacts of sample damage. To help overcome some of these issues of both sample damage and moving towards developing better operating procedures, this project focusses on attempting to establish how much laser irradiation a live cell sample can withstand before damage occurs during Raman spectroscopy in parallel with well-defined biological viability techniques; utilising MG-63 mammalian osteosarcoma cells. This will help to enable the design of robust standard operating procedures for Raman analysis in future, avoiding photodamage of live cells and delivering confidence in future Raman analysis results through reduced artefacts.
Date of Award4 Aug 2021
Original languageEnglish
Awarding Institution
  • University Of Strathclyde
SponsorsEPSRC (Engineering and Physical Sciences Research Council)
SupervisorDuncan Graham (Supervisor) & Karen Faulds (Supervisor)

Cite this

'