Skip to Content
MilliporeSigma
  • Fourier transform infrared spectroscopy imaging of live epithelial cancer cells under non-aqueous media.

Fourier transform infrared spectroscopy imaging of live epithelial cancer cells under non-aqueous media.

Journal of clinical pathology (2013-02-09)
JunYi Soh, Adeline Chueng, Aminat Adio, Alan J Cooper, Brian R Birch, Bashir A Lwaleed
ABSTRACT

Fourier transform infrared (FT-IR) imaging is increasingly being applied to biomedical specimens, but strong IR absorption by water complicates live cell imaging. This study investigates the viability of adherent epithelial cells maintained for short periods under mineral oils in order to facilitate live cell spectroscopy using FT-IR with subsequent imaging. The MGH-U1 urothelial or CaCo2 colorectal cancer cell lines were grown on plastic surfaces or mid-range infrared transparent windows. Medium in established cultures was replaced with paraffin mineral oil, or Fluorolube, for up to 2 h, and viability assessed by supravital staining. Drug handling characteristics were also assessed. Imaging of preparations was attempted by reflectance and transmission using a Varian FT-IR microscope. Cells covered by mineral oil remained viable for 2 h, with recovery into normal medium possible. MTT ((3-(4,5-dimethylthlazol-2-yl)-2,5-diphenyl tetrazolium) conversion to crystalline formazan and differential patterns of drug uptake were maintained. The combination of a calcium fluoride substrate, Fluorolube oil, and transmission optics proved best for spectroscopy. Spectral features were used to obtain images of live cells. The viability of cells overlaid with IR transparent oils was assessed as part of a technique to optimise conditions for FT-IR imaging. Images of untreated cells were obtained using both reflectance and transmission. This represents an effective means of imaging live cells by IR spectroscopy, and also means that imaging is not necessarily a terminal event. It also increases options for producing images based on real-time biochemistry in a range of in vitro experimental and 'optical biopsy' contexts.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Epirubicin hydrochloride, ≥90% (HPLC)
Epirubicin hydrochloride, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Paraffin wax, SAJ first grade, mp 58-60 °C
Sigma-Aldrich
Paraffin wax, SAJ first grade, mp 40-42 °C
Sigma-Aldrich
Paraffin wax, SAJ first grade, mp 50-52 °C
Sigma-Aldrich
Paraffin wax, SAJ first grade, mp 56-58 °C
Sigma-Aldrich
Paraffin wax, SAJ first grade, mp 64-66 °C
Sigma-Aldrich
Paraffin wax, SAJ first grade, mp 60-62 °C
Sigma-Aldrich
Paraffin wax, SAJ first grade, mp 52-54 °C
Sigma-Aldrich
Paraffin wax, SAJ first grade, mp 62-64 °C
Sigma-Aldrich
Paraffin wax, mp ≥65 °C (ASTM D 87)
Sigma-Aldrich
Paraffin wax, meets analytical specification of Ph. Eur., pastilles, white
Sigma-Aldrich
Paraffin oil, puriss., meets analytical specification of Ph. Eur., BP, viscous liquid
Sigma-Aldrich
Paraffin wax, pellets, white
Sigma-Aldrich
Paraffin wax, pellets, white, tested according to Ph. Eur.
Sigma-Aldrich
Calcium fluoride, anhydrous, powder, 99.99% trace metals basis
Sigma-Aldrich
Paraffin wax, chunks (large), white
Sigma-Aldrich
Paraffin wax, mp 53-58 °C (ASTM D 87)
Sigma-Aldrich
Paraffin wax, mp 58-62 °C (ASTM D 87)
Sigma-Aldrich
Calcium fluoride, random crystals, optical grade, 99.99% trace metals basis
Sigma-Aldrich
Paraffin wax, tested according to Ph. Eur., low viscosity
Sigma-Aldrich
Paraffin wax, tested according to Ph. Eur., high viscosity
Supelco
Paraffin oil, suitable for IR spectroscopy