Cavity enhanced absorption spectroscopy for drug quantification in living cells
Lead Research Organisation:
University of Exeter
Department Name: University of Exeter Medical School
Abstract
Aim: The aim of this project is to develop a generic label free spectroscopic method, based on cavity enhanced absorption spectroscopy, for quantifying drug intake in living cells: e.g. chemotherapy drugs entering cancer cells.
Historically drugs entering cells have been quantified by lysing the cell and measuring drug levels in the cell lysate, for example by fluorescence spectroscopy or mass spectrometry. Although this method provides important insights into the problem of drug permeation it is restricted to cell population analysis. Recently Raman Spectroscopy has been used as a detection tool to probe drug permeation directly inside cells but although promising, Raman Spectroscopy currently remains semi quantitative with the Raman signal being dependent on the complex cellular environment.
During this PhD studentship a novel Cavity-Reflection-Enhanced Light-Absorption spectrometer optimized for single cell absorption measurements will be developed. This is a miniaturized single cell spectrometer with a miniaturized detection chamber down to the cell volume and optimized for high sensitivity UV-Vis absorption measurements in cells. This platform will allow to investigate and optimise efficacy of drug intake in cells.
Workload Milestones:
1) Technology development. The PhD student taking part in this project will be involved in building and optimizing the optical setup, including fibre alignment, fibre micromachining and generating semi-transparent aluminium mirrors. Methods for separating optical signals from the drug, the cell, and the background will be developed by referencing measurements at different wavelengths ( e.g. tryptophan absorption peak at 290 nm).
2) Chemotherapy Resistance. To validate the technology for cancer research we will start by investigating the drug, doxorubicin, which enters a human lung cancerous cell-line. In this purpose COR-L23/R, a multi-drug resistant human lung cancer cell-line over expressing efflux pumps that export doxorubicin out of the cell will be compared to its parent line COR-L23. By analysing the spectral difference at the single cell level we will provide a platform to rapidly investigate the ability of doxorubicin, and other chemotherapy drugs, to permeate through the cellular membrane of cancer cells. The kinetics of cellular uptake and kinetics of resultant cell response will be investigated on this platform in multiplexed single-cell measurements. The effects of cell heterogeneity on drug efficacy will be quantified; variations of the molecular structure of a drug can will be tested in order to optimise dose and response.
Historically drugs entering cells have been quantified by lysing the cell and measuring drug levels in the cell lysate, for example by fluorescence spectroscopy or mass spectrometry. Although this method provides important insights into the problem of drug permeation it is restricted to cell population analysis. Recently Raman Spectroscopy has been used as a detection tool to probe drug permeation directly inside cells but although promising, Raman Spectroscopy currently remains semi quantitative with the Raman signal being dependent on the complex cellular environment.
During this PhD studentship a novel Cavity-Reflection-Enhanced Light-Absorption spectrometer optimized for single cell absorption measurements will be developed. This is a miniaturized single cell spectrometer with a miniaturized detection chamber down to the cell volume and optimized for high sensitivity UV-Vis absorption measurements in cells. This platform will allow to investigate and optimise efficacy of drug intake in cells.
Workload Milestones:
1) Technology development. The PhD student taking part in this project will be involved in building and optimizing the optical setup, including fibre alignment, fibre micromachining and generating semi-transparent aluminium mirrors. Methods for separating optical signals from the drug, the cell, and the background will be developed by referencing measurements at different wavelengths ( e.g. tryptophan absorption peak at 290 nm).
2) Chemotherapy Resistance. To validate the technology for cancer research we will start by investigating the drug, doxorubicin, which enters a human lung cancerous cell-line. In this purpose COR-L23/R, a multi-drug resistant human lung cancer cell-line over expressing efflux pumps that export doxorubicin out of the cell will be compared to its parent line COR-L23. By analysing the spectral difference at the single cell level we will provide a platform to rapidly investigate the ability of doxorubicin, and other chemotherapy drugs, to permeate through the cellular membrane of cancer cells. The kinetics of cellular uptake and kinetics of resultant cell response will be investigated on this platform in multiplexed single-cell measurements. The effects of cell heterogeneity on drug efficacy will be quantified; variations of the molecular structure of a drug can will be tested in order to optimise dose and response.
Organisations
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| EP/N509656/1 | 01/10/2016 | 30/09/2021 | |||
| 2241890 | Studentship | EP/N509656/1 | 01/10/2019 | 31/01/2024 | Imogen Worth |
| EP/R513210/1 | 01/10/2018 | 30/09/2023 | |||
| 2241890 | Studentship | EP/R513210/1 | 01/10/2019 | 31/01/2024 | Imogen Worth |