用于生物芯片筛选和荧光显微镜检查的共聚焦读数器。

Confocal reader for biochip screening and fluorescence microscopy.

作者信息

Ruckstuhl Thomas, Walser Andreas, Verdes Dorinel, Seeger Stefan

机构信息

Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstr. 190, CH-8057 Zürich, Switzerland.

出版信息

Biosens Bioelectron. 2005 Mar 15;20(9):1872-7. doi: 10.1016/j.bios.2004.06.050.

DOI:10.1016/j.bios.2004.06.050

PMID:15681208

Abstract

We developed a fluorescence reader for the sensitive detection of surface-generated fluorescence. The system is applicable for high resolution imaging as well as for the readout of large biochips. The surface of a microscope coverslip is scanned with a laser beam focused to a waist diameter of 500 nm (FWHM) by means of a single aspheric lens. Scanning large areas with a focused beam usually evokes the need of automatic control elements to adjust the laser spot to the designated position at the surface. Due to the special design of the reader, the focus keeps at the plane of the surface even when scanning large areas, obviating the requirement of any real time control. Thus the instrument is straightforward and inexpensive. Nevertheless it features a high sensitivity and high optical resolution. The versatility of the instrument is demonstrated by imaging cells and reading out a DNA-chip. The excellent sensitivity is shown by detecting single fluorescently labeled antibodies.

摘要

我们开发了一种用于灵敏检测表面产生荧光的荧光阅读器。该系统适用于高分辨率成像以及大型生物芯片的读数。借助单个非球面透镜，用聚焦到腰径为500纳米（半高宽）的激光束扫描显微镜盖玻片的表面。用聚焦光束扫描大面积区域通常需要自动控制元件将激光光斑调整到表面的指定位置。由于阅读器的特殊设计，即使在扫描大面积区域时，焦点也能保持在表面平面上，从而无需任何实时控制。因此，该仪器结构简单且成本低廉。然而，它具有高灵敏度和高光学分辨率。通过对细胞成像和读取DNA芯片证明了该仪器的多功能性。通过检测单个荧光标记抗体展示了其出色的灵敏度。