Laser-based Microscope Research Articles

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.

Design and Characterization of Precise Laser-Based Autofocusing Microscope with Reduced Geometrical Fluctuations

The present study proposes a laser-based microscope system in which a precise autofocusing capability is achieved using a position feedback signal based on the distance L between the geometrical center (Xc, Yc) of the image captured by the CCD sensor and the centroid (xcentroid, ycentroid) of the image. The experimental results show that the proposed system has a positioning accuracy of 2.2 μm given a working range of ±200 μm. The autofocusing performance of the proposed system is thus better than that of a conventional centroid-based system, which typically achieves a positioning accuracy of around 5.2 μm.