A novel spectral microscope system: Application in quantitative pathology
A novel spectral microscope system: Application in quantitative pathology
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DOI：
10.1109/tbme.2002.807648
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发表时间：
2003-02-01
影响因子：
4.6
通讯作者：
Balas, C
中科院分区：
文献类型：
--
作者：
Papadakis, AN;Stathopoulos, E;Balas, C
在本文中，提出了一种新型光谱显微镜系统以及一种用于定量评估病理学中使用的染色剂的组织学样本的摄取的方法，以标记具有诊断重要性的组织特征。显微镜的关键部件是基于可变干涉滤光片的单色仪。该系统能够在多个光谱带中执行实时光谱成像，并在400-1000 nm光谱范围内的任何图像像素中执行显微光谱分析。波长调谐步长为2.4-2.6 nm，而每个步长的半峰全宽约为工作中心波长的1.5%。开发的系统集成了算法和校准程序，用于计算组织的染色吸收量。从染色组织和校准染色溶液获取的光谱能够计算染色剂的浓度图，即使后者是多个并且在空间和光谱上重叠。该系统用于定量绘制乳腺癌细胞中雌激素和孕激素受体的表达。在这种特殊情况下，模型验证表明，尽管采用了两种染色剂，但需要在十多个波长下捕获它们的透射率才能获得可接受的精度。这些发现强调了在病理学中开发和实施光谱显微镜的必要性及其引入新的更可靠的诊断标准的潜力。
In this paper, a novel spectral microscope system is presented together with a method for the quantitative assessment of the uptake by histologic samples of stains used in pathology to label tissue features of diagnostic importance. The critical component of the microscope is a variable interference filter-based monochromator. The system is capable of performing real-time spectral imaging in a plurality of spectral bands and micro-spectroscopy in any image pixel, in the spectral range 400-1000 nm. The wavelength-tuning step is 2.4-2.6 nm, while the full-width at half maximum in each step is about 1.5% of the operating central wavelength. The developed system integrates algorithms and calibration procedures for the calculation of the stain-uptake by the tissue. The acquired spectra from both stained tissue and calibration stain solutions enable the calculation of the concentration maps of the stains, even if the latter are multiple and overlap spatially and spectrally. The system was used for the quantitative mapping of the expression of estrogen and progesterone receptors in breast cancer cells. In this particular case, model validation shows that although two stains are employed, capturing of their transmittance at more than ten wavelengths is required in order to obtain an, acceptable accuracy. These findings highlight the need for the development and implementation of spectral microscopy in pathology and its potential to introduce novel more reliable diagnostic criteria.