Suppr超能文献

基于光片的荧光相关光谱学中二维阵列探测器的性能

The performance of 2D array detectors for light sheet based fluorescence correlation spectroscopy.

作者信息

Singh Anand Pratap, Krieger Jan Wolfgang, Buchholz Jan, Charbon Edoardo, Langowski Jörg, Wohland Thorsten

机构信息

Department of Biological Sciences and NUS Centre for Bio-Imaging Sciences, National University of Singapore, 14 Science Drive 4, 117557, Singapore.

出版信息

Opt Express. 2013 Apr 8;21(7):8652-68. doi: 10.1364/OE.21.008652.

Abstract

Single plane illumination microscopy based fluorescence correlation spectroscopy (SPIM-FCS) is a new method for imaging FCS in 3D samples, providing diffusion coefficients, transport, flow velocities and concentrations in an imaging mode. SPIM-FCS records correlation functions over a whole plane in a sample, which requires array detectors for recording the fluorescence signal. Several types of image sensors are suitable for FCS. They differ in properties such as effective area per pixel, quantum efficiency, noise level and read-out speed. Here we compare the performance of several low light array detectors based on three different technologies: (1) Single-photon avalanche diode (SPAD) arrays, (2) passive-pixel electron multiplying charge coupled device (EMCCD) and (3) active-pixel scientific-grade complementary metal oxide semiconductor cameras (sCMOS). We discuss the influence of the detector characteristics on the effective FCS observation volume, and demonstrate that light sheet based SPIM-FCS provides absolute diffusion coefficients. This is verified by parallel measurements with confocal FCS, single particle tracking (SPT), and the determination of concentration gradients in space and time. While EMCCD cameras have a temporal resolution in the millisecond range, sCMOS cameras and SPAD arrays can extend the time resolution of SPIM-FCS down to 10 μs or lower.

摘要

基于单平面照明显微镜的荧光相关光谱技术(SPIM-FCS)是一种用于对三维样品进行荧光相关光谱成像的新方法,它能在成像模式下提供扩散系数、传输、流速和浓度等信息。SPIM-FCS记录样品中整个平面的相关函数,这需要阵列探测器来记录荧光信号。有几种类型的图像传感器适用于荧光相关光谱技术。它们在诸如每像素有效面积、量子效率、噪声水平和读出速度等特性方面存在差异。在此,我们比较了基于三种不同技术的几种低光阵列探测器的性能:(1)单光子雪崩二极管(SPAD)阵列,(2)无源像素电子倍增电荷耦合器件(EMCCD),以及(3)有源像素科学级互补金属氧化物半导体相机(sCMOS)。我们讨论了探测器特性对有效荧光相关光谱观测体积的影响,并证明基于光片的SPIM-FCS能提供绝对扩散系数。这通过与共聚焦荧光相关光谱、单粒子追踪(SPT)的平行测量以及时空浓度梯度的测定得到了验证。虽然EMCCD相机的时间分辨率在毫秒范围内,但sCMOS相机和SPAD阵列可以将SPIM-FCS的时间分辨率扩展到10微秒或更低。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验