Bassan Paul, Weida Miles J, Rowlette Jeremy, Gardner Peter
Manchester Institute of Biotechnology, The University of Manchester, 131 Princess Street, Manchester M1 7DN, UK.
Analyst. 2014 Aug 21;139(16):3856-9. doi: 10.1039/c4an00638k.
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.
振动光谱领域的化学成像正发展成为一种有前景的工具,以补充数字组织病理学。其应用包括通过基于光谱化学信息自动识别组织/细胞类型和疾病状态来筛查活检组织。为了融入临床实践,需要加快数据采集速度,以实现基于机架的系统,在该系统中,标本能够快速成像,从而与当前的可见光扫描仪竞争,可见光扫描仪能够在一夜之间扫描数百张载玻片。当前配备焦平面阵列(FPA)探测器的傅里叶变换红外(FTIR)成像,是目前用于红外吸收化学成像的最先进仪器,然而,中红外波段宽可调谐激光器的最新进展被认为是下一代显微镜最有潜力的候选者。在本文中,我们测试了一种基于量子级联激光器(QCL)的光谱成像显微镜原型,重点是离散频率化学成像。我们展示了如何在9分钟内测量一个包含200多个芯的2×2.4平方厘米乳腺组织微阵列(TMA)的酰胺I带(1655厘米-1)的蛋白质化学图像。这一结果表明,需要从几个关键波长获取化学图像的应用,将最适合使用基于激光的显微镜。