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Methodology for FTIR Imaging of Individual Cells

100%

Yao S., Cestelli Guidi M., Delugin M., Della-Ventura G., Marcelli A., Petibois C.

Acta Physica Polonica A

2016

vol. 129

issue 2

250-254

FTIR imaging is a novel spectroscopic technique able to provide cell imaging, in vivo and in real-time. However, one key issue is developing methodologies for cell culture on IR-transparent substrates fitting cell biology requirements. In this work we tested different IR-transparent substrates in terms of biotoxicity, surface properties, and spectral image acquisition qualities. Only a few substrates, namely Si₃N₄, Ge, GLS, LaF₃, Si, SrF₂, ZnS/C, ZnS/F, were found to provide cell culture conditions comparable to those observed on usual polycarbonate Petri dishes, the main limiting parameter being the toxicity of the material (ZnS, GLS, PbF₂, PbCl₂) or a poor adhesiveness (notably diamond, AgCl, CaF₂, ZnS). From substrates eligible for a good-quality cell culture, the spectral acquisition quality is mainly affected by the refractive index value. Finally, the best compromise between cell culture quality and image spectral quality could be obtained using Si and Ge substrates. This rationalization of the available IR-transparent substrates for bioimaging is particularly relevant for live cell analyses, where cell culture conditions must remain unaffected by substrate properties.

The Future of Infrared Spectroscopy in Biosciences: In Vitro, Time-Resolved, and 3D

84%

Chen H.-H., Bobroff V., Delugin M., Pineau R., Noreen R., Seydou Y., Banerjee S., Chatterjee J., Javerzat S., Petibois C.

Acta Physica Polonica A

2016

vol. 129

issue 2

255-259

Infrared (IR) spectroscopy is at the cross-roads, with the requirement to compete with cutting-edge technologies in biosciences, mostly based on analytical performances dealing with the super-resolutions: time, lateral/spatial, and contrast. IR microscopy is diffraction limited in most cases, thus not accessing to high lateral/spatial resolutions. Additionally, it has a poor signal-to-noise ratio on a single scan, thus requiring long-lasting acquisitions that are not suitable to analyze ns-lasting biochemical events. However, it is unique because it provides a broad global chemical information of the sample contents. It is also unique because it does not require heavy sample preparation nor labeling and can be coupled to other techniques (multimodality). Finally, it is again unique because it provides quantitative measurements, thus suitable for 1D to 4D data exploitation procedures. This short review shows that IR spectroscopy will be certainly subjected to a second century of innovations, maintaining its influence in the panorama of cutting-edge analytical techniques.

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2 Acta Physica Polonica A

2 2016

2 Delugin M.

2 Petibois C.

1 Banerjee S.

1 Bobroff V.

1 Cestelli Guidi M.

1 Chatterjee J.

1 Chen H.-H.

1 Della-Ventura G.

1 Javerzat S.

1 Marcelli A.

1 Noreen R.

1 Pineau R.

1 Seydou Y.

1 Yao S.

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Methodology for FTIR Imaging of Individual Cells

The Future of Infrared Spectroscopy in Biosciences: In Vitro, Time-Resolved, and 3D