Automated most-probable loss tomography of thick selectively stained biological specimens with quantitative measurement of resolution improvement

被引：27

作者：

Bouwer, JC ^{[1
]}

Mackey, MR ^{[1
]}

Lawrence, A ^{[1
]}

Deerinck, TJ ^{[1
]}

Jones, YZ ^{[1
]}

Terada, M ^{[1
]}

Martone, ME ^{[1
]}

Peltier, S ^{[1
]}

Ellisman, MH ^{[1
]}

机构：

[1] Univ Calif San Diego, Natl Ctr Microscopy & Imaging Res, Ctr Res Biol Syst, La Jolla, CA 92093 USA

来源：

JOURNAL OF STRUCTURAL BIOLOGY | 2004年 / 148卷 / 03期

关键词：

electron microscopy; tomography; most-probable loss imaging; electron energy loss spectroscopy; thick specimen imaging;

D O I：

10.1016/j.jsb.2004.08.005

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe the technique and application of energy filtering, automated most-probable loss (MPL) tomography to intermediate voltage electron microscopy (IVEM). We show that for thick, selectively stained biological specimens, this method produces a dramatic increase in resolution of the projections and the computed volumes versus standard unfiltered transmission electron microscopy (TEM) methods. This improvement in resolution is attributed to the reduction of chromatic aberration, which results from the large percentage of inelastic electron-scattering events for thick specimens. These improvements are particularly evident at the large tilt angles required to improve tomographic resolution in the z-direction. This method effectively increases the usable thickness of selectively stained samples that can be imaged at a given accelerating voltage by dramatically improving resolution versus unfiltered TEM and increasing signal-to-noise versus zero-loss imaging, thereby expanding the utility of the IVEM to deliver information from within specimens up to 3 mum thick. (C) 2004 Elsevier Inc. All rights reserved.

引用

页码：297 / 306

页数：10

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