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星形胶质细胞钙信号动力学：一种用于二维培养的低成本荧光定制系统。

Dynamics of astrocytes Ca signaling: a low-cost fluorescence customized system for 2D cultures.

作者信息

Musotto Rosa, Wanderlingh Ulderico, D'Ascola Angela, Spatuzza Michela, Catania Maria Vincenza, De Pittà Maurizio, Pioggia Giovanni

机构信息

Institute for Biomedical Research and Innovation, National Research Council (IRIB-CNR), Messina, Italy.

Department of Mathematical and Computer Sciences, Physical Sciences and Earth Sciences, University of Messina, Messina, Italy.

出版信息

Front Cell Dev Biol. 2024 Jan 23;12:1320672. doi: 10.3389/fcell.2024.1320672. eCollection 2024.

DOI:10.3389/fcell.2024.1320672
PMID:38322166
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10844566/
Abstract

In an effort to help reduce the costs of fluorescence microscopy and expand the use of this valuable technique, we developed a low-cost platform capable of visualising and analysing the spatio-temporal dynamics of intracellular Ca signalling in astrocytes. The created platform, consisting of a specially adapted fluorescence microscope and a data analysis procedure performed with Imagej Fiji software and custom scripts, allowed us to detect relative changes of intracellular Ca ions in astrocytes. To demonstrate the usefulness of the workflow, we applied the methodology to several astrocyte preparations, specifically immortalised human astrocyte cells and wild-type mouse cells. To demonstrate the reliability of the procedure, analyses were conducted by stimulating astrocyte activity with the agonist dihydroxyphenylglycine (DHPG), alone or in the presence of the antagonist 2-methyl-6-phenylethyl-pyridine (MPEP).

摘要

为了帮助降低荧光显微镜的成本并扩大这项有价值技术的应用，我们开发了一个低成本平台，该平台能够可视化和分析星形胶质细胞内钙信号的时空动态。创建的平台由一台经过特殊改装的荧光显微镜以及使用Imagej Fiji软件和自定义脚本执行的数据分析程序组成，使我们能够检测星形胶质细胞内钙离子的相对变化。为了证明该工作流程的实用性，我们将该方法应用于几种星形胶质细胞制剂，特别是永生化人星形胶质细胞和野生型小鼠细胞。为了证明该程序的可靠性，通过单独使用激动剂二羟基苯甘氨酸（DHPG）或在拮抗剂2-甲基-6-苯乙基吡啶（MPEP）存在的情况下刺激星形胶质细胞活性来进行分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/d92876de2458/fcell-12-1320672-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/25d48f3a2699/fcell-12-1320672-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/ef3e328246dd/fcell-12-1320672-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/1a1b3096d774/fcell-12-1320672-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/63e095e29a6e/fcell-12-1320672-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/82055f10c715/fcell-12-1320672-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/4a0f1ea2afa5/fcell-12-1320672-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/0b829b80e589/fcell-12-1320672-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/05f8e39c6109/fcell-12-1320672-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/d92876de2458/fcell-12-1320672-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/25d48f3a2699/fcell-12-1320672-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/ef3e328246dd/fcell-12-1320672-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/1a1b3096d774/fcell-12-1320672-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/63e095e29a6e/fcell-12-1320672-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/82055f10c715/fcell-12-1320672-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/4a0f1ea2afa5/fcell-12-1320672-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/0b829b80e589/fcell-12-1320672-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/05f8e39c6109/fcell-12-1320672-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eb5/10844566/d92876de2458/fcell-12-1320672-g009.jpg

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