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结合定量相显微镜和激光诱导冲击波用于细胞损伤研究。

Combining quantitative phase microscopy and laser-induced shockwave for the study of cell injury.

作者信息

Pouladian Pegah, Yamauchi Toyohiko, Wakida Nicole M, Gomez-Godinez Veronica, Berns Michael W, Preece Daryl

机构信息

Beckman Laser Institute, Department of Biomedical Engineering, University of California Irvine, CA 92617, USA.

Hamamatsu Photonics K. K., 5000 Hirakuchi, Hamakita, Shizuoka 434-8601, Japan.

出版信息

Biomed Opt Express. 2021 Jun 11;12(7):4020-4031. doi: 10.1364/BOE.427693. eCollection 2021 Jul 1.

Abstract

In this paper, we propose a new system for studying cellular injury. The system is a biophotonic work station that can generate Laser-Induced Shockwave (LIS) in the cell culture medium combined with a Quantitative Phase Microscope (QPM), enabling the real-time measurement of intracellular dynamics and quantitative changes in cellular thickness during the damage and recovery processes. In addition, the system is capable of Phase Contrast (PhC) and Differential Interference Contrast (DIC) microscopy. Our studies showed that QPM allows us to discern changes that otherwise would be unnoticeable or difficult to detect using phase or DIC imaging. As one application, this system enables the study of traumatic brain injury in vitro. Astrocytes are the most numerous cells in the central nervous system (CNS) and have been shown to play a role in the repair of damaged neuronal tissue. In this study, we use LIS to create a precise mechanical force in the culture medium at a controlled distance from astrocytes and measure the quantitative changes, in order of nanometers, in cell thickness. Experiments were performed in different cell culture media in order to evaluate the reproducibility of the experimental method.

摘要

在本文中,我们提出了一种用于研究细胞损伤的新系统。该系统是一个生物光子工作站,它可以在细胞培养基中产生激光诱导冲击波(LIS),并结合定量相显微镜(QPM),能够实时测量细胞内动力学以及损伤和恢复过程中细胞厚度的定量变化。此外,该系统还具备相差(PhC)显微镜和微分干涉对比(DIC)显微镜功能。我们的研究表明,定量相显微镜使我们能够辨别出那些使用相成像或微分干涉对比成像否则将无法察觉或难以检测到的变化。作为一个应用实例,该系统能够在体外研究创伤性脑损伤。星形胶质细胞是中枢神经系统(CNS)中数量最多的细胞,并且已被证明在受损神经元组织的修复中发挥作用。在本研究中,我们使用激光诱导冲击波在离星形胶质细胞一定控制距离的培养基中产生精确的机械力,并测量细胞厚度以纳米为单位的定量变化。为了评估实验方法的可重复性,在不同的细胞培养基中进行了实验。

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本文引用的文献

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Laser-Induced Shockwave (LIS) to Study Neuronal Ca Responses.激光诱导冲击波(LIS)用于研究神经元钙反应。
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Laser Scissors and Tweezers to Study Chromosomes: A Review.用于研究染色体的激光剪刀和镊子:综述
Front Bioeng Biotechnol. 2020 Jul 16;8:721. doi: 10.3389/fbioe.2020.00721. eCollection 2020.
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DNA damage induced during mitosis undergoes DNA repair synthesis.有丝分裂过程中诱导的 DNA 损伤会进行 DNA 修复合成。
PLoS One. 2020 Apr 28;15(4):e0227849. doi: 10.1371/journal.pone.0227849. eCollection 2020.
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Phagocytic response of astrocytes to damaged neighboring cells.星形胶质细胞对受损邻近细胞的吞噬反应。
PLoS One. 2018 Apr 30;13(4):e0196153. doi: 10.1371/journal.pone.0196153. eCollection 2018.
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Astrocyte roles in traumatic brain injury.星形胶质细胞在创伤性脑损伤中的作用。
Exp Neurol. 2016 Jan;275 Pt 3(0 3):305-315. doi: 10.1016/j.expneurol.2015.03.020. Epub 2015 Mar 28.

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