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用于无标记细胞识别的高光谱扫描激光光学断层扫描装置的研究。

Investigation of a hyperspectral Scanning Laser Optical Tomography setup for label-free cell identification.

作者信息

Benecke Hannes, Johannsmeier Sonja, May Tobias, Ripken Tammo

机构信息

Industrial and Biomedical Optics Department, Laser Zentrum Hannover e.V., Hollerithallee 8, 30419, Hannover, Germany.

InSCREENeX GmbH, Inhoffenstraße 7, 38124, Braunschweig, Germany.

出版信息

Sci Rep. 2024 Aug 1;14(1):17861. doi: 10.1038/s41598-024-68685-0.

Abstract

The development of non-destructive, tomographic imaging systems is a current topic of research in biomedical technologies. One of these technologies is Scanning Laser Optical Tomography (SLOT), which features a highly modular setup with various contrast mechanisms. Extending this technology with new acquisition mechanisms allows us to investigate untreated and non-stained biological samples, leaving their natural biological physiology intact. To enhance the development of SLOT, we aimed to extend the density of information with a significant increase of acquisition channels. This should allow us to investigate samples with unknown emission spectra and even allow for label-fee cell identification. We developed and integrated a hyperspectral module into an existing SLOT system. The adaptations allow for the acquisition of three-dimensional datasets containing a highly increased information density. For validation, artificial test objects were made from fluorescent acrylic and acquired with the new hyperspectral setup. In addition, measurements were made on two different human cell spheroids with an unknown spectra, to test the possibilities of label-free cell identification. The validation measurements of the artificial test target show the expected results. Furthermore, the measurements of the biological cell spheroids show small variations in their tomographic spectrum that allow for label-free cell type differentiation. The results of the biological sample demonstrate the potential of label-free cell identification of the newly developed setup.

摘要

无损断层成像系统的开发是生物医学技术当前的研究课题。其中一项技术是扫描激光光学断层扫描(SLOT),它具有高度模块化的设置和各种对比机制。用新的采集机制扩展这项技术使我们能够研究未经处理和未染色的生物样本,保持其天然生物生理状态不变。为了促进SLOT的发展,我们旨在通过显著增加采集通道来提高信息密度。这将使我们能够研究发射光谱未知的样本,甚至实现无标记细胞识别。我们开发了一个高光谱模块并将其集成到现有的SLOT系统中。这些改进使得能够采集包含高度增加的信息密度的三维数据集。为了进行验证,用荧光丙烯酸制作了人工测试物体,并使用新的高光谱设置进行采集。此外,对两种发射光谱未知的不同人类细胞球体进行了测量,以测试无标记细胞识别的可能性。人工测试目标的验证测量显示了预期结果。此外,生物细胞球体的测量显示其断层光谱存在微小差异,这使得能够进行无标记细胞类型区分。生物样本的结果证明了新开发设置进行无标记细胞识别的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/407b/11294569/487f6dc86406/41598_2024_68685_Fig1_HTML.jpg

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