Quantum cascade laser-based mid-infrared spectrochemical imaging of tissue and biofluids

Mid-infrared spectroscopic imaging is a rapidly emerging technique in biomedical research and clinical diagnostics that takes advantage of the unique molecular fingerprint of cells, tissue and biofluids to provide a rich biochemical image without the need for staining. Spectroscopic analysis allows for the objective classification of biological material at a molecular level.1 This “label free” molecular imaging technique has been applied to histology, cytology, surgical pathology, microbiology and stem cell research, and can be used to detect subtle changes to the genome, proteome and metabolome.2,3,4 The new wealth of biochemical information made available by this technique has the distinct potential to improve cancer patient outcome through the identification of earlier stages of disease, drug resistance, new disease states and high-risk populations.4 However, despite the maturity of this science, instrumentation that provide increased sample throughput, improved image quality, a small footprint, low maintenance and require minimal spectral expertise are essential for clinical translation

Introducing discrete frequency infrared technology for high-throughput biofluid screening

Accurate early diagnosis is critical to patient survival, management and quality of life. Biofluids are key to early diagnosis due to their ease of collection and intimate involvement in human function. Large-scale mid-IR imaging of dried fluid deposits offers a high-throughput molecular analysis paradigm for the biomedical laboratory. The exciting advent of tuneable quantum cascade lasers allows for the collection of discrete frequency infrared data enabling clinically relevant timescales. By scanning targeted frequencies spectral quality, reproducibility and diagnostic potential can be maintained while significantly reducing acquisition time and processing requirements, sampling 16 serum spots with 0.6, 5.1 and 15% relative standard deviation (RSD) for 199, 14 and 9 discrete frequencies respectively. We use this reproducible methodology to show proof of concept rapid diagnostics; 40 unique dried liquid biopsies from brain, breast, lung and skin cancer patients were classified in 2.4 cumulative seconds against 10 non-cancer controls with accuracies of up to 90%

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.</p

Mid-infrared metabolic imaging with vibrational probes.

Understanding metabolism is indispensable in unraveling the mechanistic basis of many physiological and pathological processes. However, in situ metabolic imaging tools are still lacking. Here we introduce a framework for mid-infrared (MIR) metabolic imaging by coupling the emerging high-information-throughput MIR microscopy with specifically designed IR-active vibrational probes. We present three categories of small vibrational tags including azide bond, 13C-edited carbonyl bond and deuterium-labeled probes to interrogate various metabolic activities in cells, small organisms and mice. Two MIR imaging platforms are implemented including broadband Fourier transform infrared microscopy and discrete frequency infrared microscopy with a newly incorporated spectral region (2,000-2,300 cm-1). Our technique is uniquely suited to metabolic imaging with high information throughput. In particular, we performed single-cell metabolic profiling including heterogeneity characterization, and large-area metabolic imaging at tissue or organ level with rich spectral information