Universidad de Murcia, Laboratorio de Optica, Campus de Espinardo (CiOyN), Murcia, 30100 Spain.
J Biomed Opt. 2010 Mar-Apr;15(2):026007. doi: 10.1117/1.3369001.
A multiphoton microscope incorporating a Hartmann-Shack (HS) wavefront sensor to control the ultrafast laser beam's wavefront aberrations has been developed. This instrument allowed us to investigate the impact of the laser beam aberrations on two-photon autofluorescence imaging of human retinal tissues. We demonstrated that nonlinear microscopy images are improved when laser beam aberrations are minimized by realigning the laser system cavity while wavefront controlling. Nonlinear signals from several human retinal anatomical features have been detected for the first time, without the need of fixation or staining procedures. Beyond the improved image quality, this approach reduces the required excitation power levels, minimizing the side effects of phototoxicity within the imaged sample. In particular, this may be important to study the physiology and function of the healthy and diseased retina.
我们开发了一种结合哈特曼-夏克（HS）波前传感器的多光子显微镜，以控制超快速激光光束的波前像差。该仪器使我们能够研究激光光束像差对人视网膜组织的双光子自发荧光成像的影响。我们证明，通过在波前控制的同时重新调整激光系统腔，可以最小化激光光束像差，从而改善非线性显微镜图像。我们首次检测到来自人视网膜几种解剖特征的非线性信号，而无需固定或染色程序。除了提高图像质量外，这种方法还降低了所需的激发功率水平，最大限度地减少了被成像样本中光毒性的副作用。特别是，这对于研究健康和患病视网膜的生理学和功能可能很重要。
