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用于干细胞成脂分化超快速无标记监测的自发荧光-拉曼映射集成分析

Autofluorescence-Raman Mapping Integration analysis for ultra-fast label-free monitoring of adipogenic differentiation of stem cells.

作者信息

Suhito Intan Rosalina, Han Yoojoong, Ryu Yong-Sang, Son Hyungbin, Kim Tae-Hyung

机构信息

School of Integrative Engineering, Chung-Ang University, Seoul 06974, South Korea.

R&D Division, Nanobase, Inc., Seoul, South Korea.

出版信息

Biosens Bioelectron. 2021 Apr 15;178:113018. doi: 10.1016/j.bios.2021.113018. Epub 2021 Jan 27.

Abstract

Stem cell-based therapies have recently emerged to treat various incurable diseases and disorders. Types of stem cell-derived cells and their functions should be intensively analyzed before therapy. However, current pre-treatment steps for biological analysis are mostly destructive. Here, we report a novel optical method that enables ultra-fast and label-free characterization of cells, eliminating invasive, destructive steps. The technique, referred to as "autofluorescence-Raman mapping integration (ARMI)" analysis uses cell autofluorescence (AF) to reveal cellular morphology and cytosolic microstructures, while Raman mapping allows site-specific intensive analysis of target molecules, which enables ultra-fast identification of cell types. We used human mesenchymal stem cells (MSCs) as a model and induced adipogenesis. Lipid droplets in cells appeared as "blanks" in three-dimensional AF images and site-specific Raman mapping guided by AF identified the structure and components of the CH stretch. Adipogenesis could be rapidly and precisely analyzed, not only for the same batch but also for different batches. Therefore, the developed tool is highly useful for the accurate screening of stem cell differentiation and implementation in biomedical and clinical applications.

摘要

基于干细胞的疗法最近已出现,用于治疗各种无法治愈的疾病和病症。在治疗前,应深入分析干细胞衍生细胞的类型及其功能。然而,当前用于生物学分析的预处理步骤大多具有破坏性。在此,我们报告了一种新型光学方法,该方法能够对细胞进行超快速且无标记的表征,消除了侵入性、破坏性步骤。这项技术被称为“自发荧光-拉曼映射整合(ARMI)”分析,它利用细胞自发荧光(AF)来揭示细胞形态和胞质微观结构,而拉曼映射则允许对目标分子进行位点特异性的深入分析,从而能够超快速识别细胞类型。我们使用人间充质干细胞(MSCs)作为模型并诱导其成脂分化。细胞中的脂滴在三维AF图像中呈现为“空白”,由AF引导的位点特异性拉曼映射确定了CH伸缩振动的结构和成分。不仅对于同一批次,而且对于不同批次,都能够快速且精确地分析成脂分化情况。因此,所开发的工具对于准确筛选干细胞分化以及在生物医学和临床应用中的实施非常有用。

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