University of Florence, Department of Physics and Astronomy, Florence, Italy.
European Laboratory for Non-Linear Spectroscopy, Sesto Fiorentino, Italy.
J Biomed Opt. 2024 Sep;29(9):093508. doi: 10.1117/1.JBO.29.9.093508. Epub 2024 Sep 10.
Histopathological examination of surgical biopsies, such as in glioma and glioblastoma resection, is hindered in current clinical practice by the long time required for the laboratory analysis and pathological screening, typically taking several days or even weeks to be completed.
We propose here a transportable, high-density, spectral scanning-based hyperspectral imaging (HSI) setup, named HyperProbe1, that can provide , fast biochemical analysis, and mapping of fresh surgical tissue samples, right after excision, and without the need for fixing, staining nor compromising the integrity of the tissue properties.
HyperProbe1 is based on spectral scanning via supercontinuum laser illumination filtered with acousto-optic tunable filters. Such methodology allows the user to select any number and type of wavelength bands in the visible and near-infrared range between 510 and 900 nm (up to a maximum of 79) and to reconstruct 3D hypercubes composed of high-resolution (4 to ), widefield images ( ) of the surgical samples, where each pixel is associated with a complete spectrum.
The HyperProbe1 setup is here presented and characterized. The system is applied to 11 fresh surgical biopsies of glioma from routine patients, including different grades of tumor classification. Quantitative analysis of the composition of the tissue is performed via fast spectral unmixing to reconstruct the mapping of major biomarkers, such as oxy-( ) and deoxyhemoglobin (HHb), as well as cytochrome-c-oxidase (CCO). We also provided a preliminary attempt to infer tumor classification based on differences in composition in the samples, suggesting the possibility of using lipid content and differential CCO concentrations to distinguish between lower and higher-grade gliomas.
A proof of concept of the performances of HyperProbe1 for quantitative, biochemical mapping of surgical biopsies is demonstrated, paving the way for improving current post-surgical, histopathological practice via non-destructive, streamlined screening of fresh tissue samples in a matter of minutes after excision.
在当前的临床实践中，手术活检（如在胶质瘤和胶质母细胞瘤切除术中）的组织病理学检查受到实验室分析和病理筛选所需时间长的阻碍，通常需要数天甚至数周才能完成。
我们在这里提出了一种可移植的、高密度的、基于光谱扫描的高光谱成像（HSI）设备，命名为 HyperProbe1，它可以在切除后立即对新鲜手术组织样本进行快速生化分析和映射，而无需固定、染色，也不会损害组织特性的完整性。
HyperProbe1 基于超连续激光照明的光谱扫描，通过声光可调谐滤波器进行过滤。这种方法允许用户在 510 到 900nm 之间的可见和近红外范围内选择任意数量和类型的波长带（最多可达 79 个），并重建由高分辨率（4 到 ）组成的 3D 超立方体，宽场图像（ ）的手术样本，其中每个像素都与一个完整的光谱相关联。
介绍并描述了 HyperProbe1 设备。该系统应用于 11 例来自常规患者的胶质瘤新鲜手术活检，包括不同等级的肿瘤分类。通过快速光谱解混对组织成分进行定量分析，以重建主要生物标志物（如氧合血红蛋白（ ）和脱氧血红蛋白（HHb））以及细胞色素-c-氧化酶（CCO）的映射。我们还初步尝试根据样本成分的差异推断肿瘤分类，表明使用脂质含量和差异 CCO 浓度来区分低级别和高级别胶质瘤的可能性。
证明了 HyperProbe1 用于手术活检的定量、生化映射的概念验证，为通过非破坏性、几分钟内对切除后的新鲜组织样本进行简化筛选，改进当前的术后组织病理学实践铺平了道路。
