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一种用于活细胞Fura-2 AM比率钙成像的340/380纳米发光二极管照明器，精度优于5纳摩尔。

A 340/380 nm light-emitting diode illuminator for Fura-2 AM ratiometric Ca imaging of live cells with better than 5 nM precision.

作者信息

Tinning P W, Franssen A J P M, Hridi S U, Bushell T J, McConnell G

机构信息

Department of Physics, SUPA University of Strathclyde, Glasgow, U.K.

Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, U.K.

出版信息

J Microsc. 2018 Mar;269(3):212-220. doi: 10.1111/jmi.12616. Epub 2017 Aug 24.

DOI:10.1111/jmi.12616
PMID:28837217
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5836901/
Abstract

We report the first demonstration of a fast wavelength-switchable 340/380 nm light-emitting diode (LED) illuminator for Fura-2 ratiometric Ca imaging of live cells. The LEDs closely match the excitation peaks of bound and free Fura-2 and enables the precise detection of cytosolic Ca concentrations, which is only limited by the Ca response of Fura-2. Using this illuminator, we have shown that Fura-2 acetoxymethyl ester (AM) concentrations as low as 250 nM can be used to detect induced Ca events in tsA-201 cells and while utilising the 150 μs switching speeds available, it was possible to image spontaneous Ca transients in hippocampal neurons at a rate of 24.39 Hz that were blunted or absent at typical 0.5 Hz acquisition rates. Overall, the sensitivity and acquisition speeds available using this LED illuminator significantly improves the temporal resolution that can be obtained in comparison to current systems and supports optical imaging of fast Ca events using Fura-2.

摘要

我们报告了首个用于活细胞Fura-2比率钙成像的快速波长可切换340/380 nm发光二极管（LED）照明器的演示。这些LED与结合态和游离态Fura-2的激发峰紧密匹配，能够精确检测胞质钙浓度，而这仅受Fura-2的钙响应限制。使用该照明器，我们发现低至250 nM的Fura-2乙酰氧基甲酯（AM）浓度可用于检测tsA-201细胞中诱导的钙事件，并且利用150 μs的切换速度，可以以24.39 Hz的速率对海马神经元中的自发钙瞬变进行成像，而在典型的0.5 Hz采集速率下这些瞬变会减弱或消失。总体而言，与当前系统相比，使用该LED照明器可获得的灵敏度和采集速度显著提高了时间分辨率，并支持使用Fura-2对快速钙事件进行光学成像。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/b9350c0c25d4/JMI-269-212-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/f81c56e69254/JMI-269-212-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/102bcc174aba/JMI-269-212-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/a22320249f8e/JMI-269-212-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/d6d8efa544cb/JMI-269-212-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/a1dbb83a59f1/JMI-269-212-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/000848ae737a/JMI-269-212-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/b9350c0c25d4/JMI-269-212-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/f81c56e69254/JMI-269-212-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/102bcc174aba/JMI-269-212-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/a22320249f8e/JMI-269-212-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/d6d8efa544cb/JMI-269-212-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/a1dbb83a59f1/JMI-269-212-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/000848ae737a/JMI-269-212-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/21f9/5836901/b9350c0c25d4/JMI-269-212-g007.jpg

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