激光扫描共聚焦显微镜_图文讲解

激光扫描共聚焦显微镜

吴旭

2008.10.14

高级显微镜原理

正置、倒置显微镜

细胞遗传工作站

活细胞工作站

激光显微分离系统

激光共聚焦显微镜

概述

激光扫描共聚焦显微镜

（

Laser scanning confocalmicroscope

，

LSCM

）

生物医学领域的主要应用

通过一种或 者多种荧光探针标记后，可对固定的组织或活体样本进行亚细胞水

平结构功能研究

高空间分辨率、非介入无损伤连续光学切片、三维图像、实时动态等 细胞结构

和功能的分析检测

……

Conventional fluorescence microscope Confocal

microscope

历史

1957

年，

Marvin Mins ky

提出了共聚焦显微镜技术的某些基本原理，获得了美

国的专 利。

1967

年，

Egger

和

Petran

成功地应用共聚焦显微镜产生了一个光学横断面。

1977

年，

Sheppard

和

Wilson

首次描述了光与被照 明物体的原子之间的非线性

关系和激光扫描器的拉曼光谱学。

1984

年，

Biorad

为公司推出了世界第一台商品化的共聚焦显微镜，型号为

SOM-100

，扫描方式为台阶式扫描。

1986

年

MRC-500

型改进为光束扫描，用作生

< br>物荧光显微镜的共聚焦系统。

Confocal microscopy comes of ageJG White & WB Amos. Nature 328, 183 -184

(09 July 1987

Zeiss

、

Leica

、

Meridian

、

Olympus

Zeiss LSM510

激光扫描共聚焦显微镜

Zeiss LSM510 META

激光扫描共聚焦显微镜

Zeiss LSM510 META

激光扫描共聚焦显微镜

Nikon A1R

激光扫描共聚焦显微镜

Prairie UltimaIV

活体双光子显微镜

国家光电实验室（武汉）自制随机定位双光子显微镜

Leica TCS SP5

激光共聚焦扫描显微镜

基本原理

相差、

DIC

常用荧光标记

共聚焦原理

Two ways to obtain contrast in light microscopy. The stained portions of the cell in

(A reduce the amplitude of light waves of particular wavelengths passing through

them.

A colored image of the cell is thereby obtained that is visible in the ordinary way.

Light passing through the unstained, living cell (B undergoes very little change in

amplitude, and the structural details cannot be seen even if the image is highly magnified.

The phase of the light, however, is altered by its passage through the cell, and small

phase differences can be made visible by exploiting interference effects using a phase-

contrast or a differential-interference-contrast microscope.

D. Phase-contrast or a

differential-interference-

contrast microscope

Four types of light microscopy. (A The image of a fibroblast in culture obtained by

the simple transmission of light through the cell, atechnique known as bright-field

other images were obtained by techniques discussed in the text: (B

phase-contrast microscopy, (C Nomarski differential-interference-contrast microscopy,

and (D dark-field microscopy.

常用荧光探针

Proteins Nucleic Acids DNA Ions pH Sensitive Indicators Oxidation States Specific

Organelles