Large scale infrared imaging of tissue micro arrays (TMAs) using a tunable Quantum Cascade Laser (QCL) based microscope.

Chemical imaging in the field of vibrational spectroscopy is developing into a promising tool to complement digital histopathology. Applications include screening of biopsy tissue via automated recognition of tissue/cell type and disease state based on the chemical information from the spectrum. For integration into clinical practice, data acquisition needs to be speeded up to implement a rack based system where specimens are rapidly imaged to compete with current visible scanners where 100's of slides can be scanned overnight. Current Fourier transform infrared (FTIR) imaging with focal plane array (FPA) detectors are currently the state-of-the-art instrumentation for infrared absorption chemical imaging, however recent development in broadly tunable lasers in the mid-IR range is considered the most promising potential candidate for next generation microscopes. In this paper we test a prototype quantum cascade laser (QCL) based spectral imaging microscope with a focus on discrete frequency chemical imaging. We demonstrate how a protein chemical image of the amide I band (1655 cm(-1)) of a 2 × 2.4 cm(2) breast tissue microarray (TMA) containing over 200 cores can be measured in 9 min. This result indicates that applications requiring chemical images from a few key wavelengths would be ideally served by laser-based microscopes.

[1]  Noel W Clarke,et al.  Whole organ cross-section chemical imaging using label-free mega-mosaic FTIR microscopy. , 2013, The Analyst.

[2]  Peter Lasch,et al.  Biomedical Vibrational Spectroscopy , 2008 .

[3]  Hugh J. Byrne,et al.  Resonant Mie scattering (RMieS) correction of infrared spectra from highly scattering biological samples. , 2010, The Analyst.

[4]  Rohit Bhargava,et al.  Discrete frequency infrared microspectroscopy and imaging with a tunable quantum cascade laser. , 2012, Analytical chemistry.

[5]  N. Clarke,et al.  FTIR microscopy of biological cells and tissue: data analysis using resonant Mie scattering (RMieS) EMSC algorithm. , 2012, The Analyst.

[6]  Virgilia Macias,et al.  High-resolution Fourier-transform infrared chemical imaging with multiple synchrotron beams , 2011, Nature Methods.

[7]  Rohit Bhargava,et al.  Mid-infrared microspectroscopic imaging with a quantum cascade laser , 2013, Defense, Security, and Sensing.

[8]  Ehsan Gazi,et al.  A correlation of FTIR spectra derived from prostate cancer biopsies with gleason grade and tumour stage. , 2006, European urology.

[9]  Faraday Discuss , 1985 .

[10]  Jaleel A. Miyan,et al.  The combined application of FTIR microspectroscopy and ToF-SIMS imaging in the study of prostate cancer. , 2004, Faraday discussions.

[11]  Peter Gardner,et al.  Automated high-throughput assessment of prostate biopsy tissue using infrared spectroscopic chemical imaging , 2014, Medical Imaging.

[12]  J Dwyer,et al.  Applications of Fourier transform infrared microspectroscopy in studies of benign prostate and prostate cancer. A pilot study , 2003, The Journal of pathology.

[13]  B. Krauskopf,et al.  Proc of SPIE , 2003 .

[14]  ROHIT BHARGAVA,et al.  Infrared Spectroscopic Imaging: The Next Generation , 2012, Applied spectroscopy.

[15]  Paul Bassan,et al.  Transmission FT-IR chemical imaging on glass substrates: applications in infrared spectral histopathology. , 2014, Analytical chemistry.

[16]  S. Hewitt,et al.  Infrared spectroscopic imaging for histopathologic recognition , 2005, Nature Biotechnology.

[17]  N. Clarke,et al.  FTIR-based spectroscopic analysis in the identification of clinically aggressive prostate cancer , 2008, British Journal of Cancer.

[18]  Paul Dumas,et al.  Resonant Mie scattering in infrared spectroscopy of biological materials--understanding the 'dispersion artefact'. , 2009, The Analyst.

[19]  Rohit Bhargava,et al.  High throughput assessment of cells and tissues: Bayesian classification of spectral metrics from infrared vibrational spectroscopic imaging data. , 2006, Biochimica et biophysica acta.