Correlative In Vivo 2 Photon and Focused Ion Beam Scanning Electron Microscopy of Cortical Neurons

被引：60

作者：

Maco, Bohumil ^{[1
,2
]}

Holtmaat, Anthony ^{[3
]}

Cantoni, Marco ^{[1
]}

Kreshuk, Anna ^{[4
]}

Straehle, Christoph N. ^{[4
]}

Hamprecht, Fred A. ^{[4
]}

Knott, Graham W. ^{[1
]}

机构：

[1] Ecole Polytech Fed Lausanne, BioEM Facil, Ctr Electron Microscopy, CH-1015 Lausanne, Switzerland

[2] Ecole Polytech Fed Lausanne, CV Lab, CH-1015 Lausanne, Switzerland

[3] Univ Geneva, Univ Med Ctr CMU, Dept Neurosci, Geneva, Switzerland

[4] Heidelberg Univ, Heidelberg Collaboratory Image Proc HCI, Heidelberg, Germany

来源：

PLOS ONE | 2013年 / 8卷 / 02期

基金：

瑞士国家科学基金会;

关键词：

TISSUE; LIGHT; GFP;

D O I：

10.1371/journal.pone.0057405

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Correlating in vivo imaging of neurons and their synaptic connections with electron microscopy combines dynamic and ultrastructural information. Here we describe a semi-automated technique whereby volumes of brain tissue containing axons and dendrites, previously studied in vivo, are subsequently imaged in three dimensions with focused ion beam scanning electron microcopy. These neurites are then identified and reconstructed automatically from the image series using the latest segmentation algorithms. The fast and reliable imaging and reconstruction technique avoids any specific labeling to identify the features of interest in the electron microscope, and optimises their preservation and staining for 3D analysis.

引用

页数：7

共 17 条

[1]

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