Infrared spectroscopic imaging has emerged as a powerful label-free diagnostic tool to study the molecular composition and organization in biological tissues and cells. We report infrared spectroscopic imaging using polarized light to study differential absorption of plane-polarized light by an oriented sample to detect valuable information, such as, birefringence and dichroism. For instance, the organization of collagen, specifically fiber orientation and alignment, is crucial in understanding the progression and metastasis of cancer. Recent advancements in the development of Quantum Cascade Lasers (QCL) sources have opened new avenues for high SNR measurements in the field of IR spectroscopy. In addition, QCL sources are intrinsically polarized and orientation information can be obtained at discrete frequencies with different polarization orientations, allowing much faster acquisition than a corresponding FT-IR approach. We demonstrate improved performance in terms of fast and comprehensive polarimetric image acquisition and analysis using custom-built QCL microscope and evaluate its impact on applications by analyzing the important spectral bands of surgical tissue sections.
Infrared (IR) spectroscopic imaging is an emerging modality for biological tissue analysis that has traditionally employed an interferometer for spectral discrimination. Recent technology developments have made discrete frequency sources, both lasers and filters, practical for imaging. The use of quantum cascade lasers in particular, presents new opportunities as well as challenges. Here we describe results from a novel point scanning confocal IR microscope and demonstrate the performance imaging several important spectral bands of lung tissue. Results show the possibility of discrete frequency (DF) absorbance measurements with RMS noise levels down to 0.34 mAU in 0.25 ms.
Conventional mid-infrared (mid-IR) Fourier transform infrared (FT-IR) spectroscopic imaging systems employ an incoherent globar source and achieve spectral contrast through interferometry. While this approach is suitable for many general applications, recent advancements in broadly tunable external cavity Quantum Cascade Lasers (QCL) offer new approaches to and new possibilities for mid-IR micro-spectroscopic imaging. While QCL-based devices have yet to achieve the wide spectral range generally employed by spectroscopists for molecular analyses, they are starting to be used for microscopy at discrete frequencies. Here, we present a discrete frequency IR (DFIR) microscope based on a QCL source and explore its utility for mid-IR imaging. In our prototype instrument, spectral contrast is achieved by tuning the QCL to bands in a narrow spectral region of interest. We demonstrate wide-field imaging employing a 128x128 pixel liquid nitrogen cooled mercury cadmium telluride (MCT) focal plane array (FPA) detector. The resulting images demonstrate successful imaging as well as several unique features due to coherence effects from the laser source. Here we discuss the effects of this coherence and compare our instrument to conventional mid-IR imaging instrumentation.