光谱学与光谱分析, 2009, 29 (9): 2330, 网络出版: 2010-05-26  

基于激光共焦扫描术的亚心形扁藻显微图像与光谱

Fluorescence Spectra and Imaging of Platymonas Subcordiformis via LSCM
作者单位
1 福建师范大学, 医学光电科学与技术教育部重点实验室, 福建 福州350007
2 华中科技大学光电子科学与工程学院, 湖北 武汉430074
3 福建师范大学生命科学学院, 福建 福州350007
摘要
采用激光共聚焦扫描显微技术, 针对亚心形扁藻开展了研究。 从获得的488 nm Ar+激光单光子激发的亚心形扁藻自体荧光光谱与图像, 可知细胞内有一杯状叶绿体物质, 其荧光峰值为682 nm, 对应叶绿体发出的红色荧光。 在单通道模式下, 获得800 nm fs激光双光子激发的扁藻自体荧光光谱与图像, 可知每个杯状叶绿体的内部有一个自体荧光更强的圆形物质。 在双通道模式下, 可分别获得小圆形物质的自体荧光图像, 杯状叶绿体自体荧光图像, 以及两个通道图像的叠加。 进一步获得了双光子藻细胞荧光图的6个主要的荧光峰。 采用单光子激光激发可获得亚心形扁藻叶绿体自体荧光图像及其荧光光谱, 而双光子激光激发荧光光谱的多通道以及Lambda模式下采集光谱信号与图像, 不仅可观察到亚心形扁藻的内部形态结构, 还可能从双光子激发荧光图中研究分析亚心形扁藻生化物质的存在, 灵敏度较单光子激发高。 激光扫描共聚焦显微技术, 特别是双光子荧光与图像技术可为海藻的检测与研究提供一种快速、 实时、 有效、 简便的方法。
Abstract
Primary investigation of Platymonas subcordiformis was performed with laser scanning microscopic technology. Both autofluorescence spectra and autofluorescence imaging of Platymonas subcordiformis were achieved by one photon excitation of 488 nm Ar+ laser. Autofluorescence images revealed cup-shaped object inside the Platymonas subcordiformis cell, and the corresponding fluorescence peak located at 682 nm was attributed to chloroplast. Moreover, in single channel mode, there existed a round-shaped object in chloroplast center, showing strong fluorescence by 800 nm femtosecond laser excitation. Also, the authors obtained both the individual cup-shaped chloroplast and round-shaped object image and the overlaid images by using dual-channel mode. Meanwhile, six main fluorescence peaks were found. Single photon excitation can be used to obtain autofluorescence image and autofluorescence spectra of Platymonas subcordiformis, while two photon excitation combined with multitrack mode and Lambda mode can be used not only to observe internal structure, but also for the analysis of existence of biological and chemical substances with higher sensitivity than single photon excitation. LSCM technique can be a promising tool for rapid, convenient, real-time and effective investigation in ocean algae.

陈冠楠, 黄祖芳, 陈荣, 林居强, 陈建新, 杨坤涛, 庄惠如, 王丽姜. 基于激光共焦扫描术的亚心形扁藻显微图像与光谱[J]. 光谱学与光谱分析, 2009, 29(9): 2330. CHEN Guan-nan, HUANG Zu-fang, CHEN Rong, LIN Ju-qiang, CHEN Jian-xin, YANG Kun-tao, ZHUANG Hui-ru, WANG Li-jiang. Fluorescence Spectra and Imaging of Platymonas Subcordiformis via LSCM[J]. Spectroscopy and Spectral Analysis, 2009, 29(9): 2330.

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