光学学报, 2012, 32 (10): 1018001, 网络出版: 2012-07-17
多变视场的多焦点多光子激发荧光显微技术
Multiple Variational Field-of-View Multifocal Multiphoton Fluorescence Microscope
摘要
发展了一种新型的多焦点多光子激发荧光显微技术,通过软件控制空间光调制器,在所需要的成像区域产生相应的激发光点阵,通过扫描入射角实现激发点阵快速并行扫描激发,通过CCD并行记录所产生的荧光信号,获得任意视场图像。分别实现了10×10和50×50点阵激发下的全视场双光子荧光图像,并且实现了同时多个区域寻址的多焦点激发成像。对比其他多焦点显微技术,该技术具有明显的灵活性,不但保持了多焦点激发的快速成像的优点,而且还增加了任意数量成像区域同时寻址和阵列点密度变化,所有这些变化只需要通过软件加载相应的相位图案给空间光调制器,而不需要任何硬件更改。
Abstract
A novel multifocal multiphoton excited fluourescence microscopy (MMM) is presented. The required focus array can be produced in multiple interesting subregions through uploading the accurate phase pattern to the spatial light modulator (SLM) with the software. The focus array can be scanned with a two-mirror galvo scanner by scanning the single beam that is incident on the SLM. The two photon fluorescence signal is recorded directly with an electron-multiplying CCD in parallel. The two photon fluorescence images in the whole field-of-view are obtained under the different modes such as 10×10 and 50×50 arrays, and the multiple subregions of interest are addressably excited simultaneously. Compared with the other MMMs, our MMM has a good flexibility without sacrificing of the imaging speed and spatial resolution and also without hardware changes.
李震, 屈军乐, 邵永红, 高志. 多变视场的多焦点多光子激发荧光显微技术[J]. 光学学报, 2012, 32(10): 1018001. Li Zhen, Qu Junle, Shao Yonghong, Gao Zhi. Multiple Variational Field-of-View Multifocal Multiphoton Fluorescence Microscope[J]. Acta Optica Sinica, 2012, 32(10): 1018001.