Quantitative imaging in the laboratory: Fast kinetics and fluorescence quenching
Quantitative imaging in the laboratory: Fast kinetics and fluorescence quenching
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DOI：
10.1021/ed084p1319
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发表时间：
2007-08-01
影响因子：
3
通讯作者：
Hanley, Quentin S.
中科院分区：
文献类型：
--
作者：
Cumberbatch, Tanya;Hanley, Quentin S.
将成像技术引入化学和生化分析已经彻底改变了原子光谱和药物发现等多个领域。传统上，科学相机和相关分析软件价格昂贵、难以使用，并且在教学实验室中并未广泛使用。尽管成像技术在化学、生物学、生物化学、生物物理学和医学的许多领域广泛使用，但它们在本科生实验室中并不广泛。本文描述了一个基于定量成像的简单、廉价的实验。该实验允许学生使用荧光素双阴离子猝灭来测量接近 2 x 109L mol-1s 的速率常数。该实验已在四个学年中由 75 名学生在中级生物化学课程中使用。在这种背景下，它被用来向学生介绍荧光猝灭的原理，荧光猝灭在现代生物物理化学中以多种方式使用。它也可能是物理化学实验室模块的合适平台。
The introduction of imaging technologies into chemical and biochemical analysis has revolutionized fields as diverse as atomic spectroscopy and drug discovery. Traditionally, scientific cameras and associated analysis software have been expensive, difficult to use, and not widely distributed in teaching laboratories. Despite the widespread use of imaging technologies in many areas of chemistry, biology, biochemistry, biophysics, and medicine, they are not widely found in the undergraduate laboratory. This article describes a simple, inexpensive, experiment based on quantitative imaging. The experiment allows students to use quenching of fluorescein dianion to measure rate constants near 2 x 109L mol-1s. This experiment has been used during four academic years in an intermediate biochemistry course by ∼75 students. In this context, it was used to introduce students to the principles of fluorescence quenching, which is used in a variety of ways in modern biophysical chemistry. It may also be a suitable platform for lab modules for physical chemistry.