深部组织双光子显微镜检查

Deep tissue two-photon microscopy.

作者信息

Helmchen Fritjof, Denk Winfried

机构信息

Department of Neurophysiology, Brain Research Institute, University of Zurich, CH-8057 Zurich, Switzerland.

出版信息

Nat Methods. 2005 Dec;2(12):932-40. doi: 10.1038/nmeth818.

DOI:10.1038/nmeth818

PMID:16299478

Abstract

With few exceptions biological tissues strongly scatter light, making high-resolution deep imaging impossible for traditional-including confocal-fluorescence microscopy. Nonlinear optical microscopy, in particular two photon-excited fluorescence microscopy, has overcome this limitation, providing large depth penetration mainly because even multiply scattered signal photons can be assigned to their origin as the result of localized nonlinear signal generation. Two-photon microscopy thus allows cellular imaging several hundred microns deep in various organs of living animals. Here we review fundamental concepts of nonlinear microscopy and discuss conditions relevant for achieving large imaging depths in intact tissue.

摘要

除了少数例外情况，生物组织会强烈散射光，这使得传统的（包括共聚焦）荧光显微镜无法进行高分辨率的深度成像。非线性光学显微镜，特别是双光子激发荧光显微镜，克服了这一限制，实现了较大的深度穿透，主要是因为即使是多次散射的信号光子也可以由于局部非线性信号产生而被追溯到其来源。因此，双光子显微镜能够对活体动物各种器官中几百微米深处的细胞进行成像。在这里，我们回顾非线性显微镜的基本概念，并讨论在完整组织中实现大成像深度的相关条件。