Benninger Richard K P, Piston David W
University of Colorado, Anschutz Medical Campus, Aurora, Colorado.
Vanderbilt University Medical Center, Nashville, Tennessee.
Curr Protoc Cell Biol. 2013 Jun;Chapter 4:4.11.1-4.11.24. doi: 10.1002/0471143030.cb0411s59.
Two-photon excitation microscopy is an alternative to confocal microscopy that provides advantages for three-dimensional and deep tissue imaging. This unit will describe the basic physical principles behind two-photon excitation and discuss the advantages and limitations of its use in laser-scanning microscopy. The principal advantages of two-photon microscopy are reduced phototoxicity, increased imaging depth, and the ability to initiate highly localized photochemistry in thick samples. Practical considerations for the application of two-photon microscopy will then be discussed, including recent technological advances. This unit will conclude with some recent applications of two-photon microscopy that highlight the key advantages over confocal microscopy and the types of experiments which would benefit most from its application.
双光子激发显微镜是共聚焦显微镜的一种替代方法，在三维和深层组织成像方面具有优势。本单元将描述双光子激发背后的基本物理原理，并讨论其在激光扫描显微镜中应用的优缺点。双光子显微镜的主要优点是光毒性降低、成像深度增加以及能够在厚样品中引发高度局部化的光化学反应。随后将讨论双光子显微镜应用的实际考虑因素，包括最近的技术进展。本单元将以双光子显微镜的一些最新应用作为结尾，这些应用突出了其相对于共聚焦显微镜的关键优势以及最能从其应用中受益的实验类型。
