Fluorescence and SEM correlative microscopy for nanomanipulation of subcellular structures

Zheng Gong; Brandon K Chen; Jun Liu; Chao Zhou; Dave Anchel; Xiao Li; Ji Ge; David P Bazett-Jones; Yu Sun

doi:10.1038/lsa.2014.105

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Fluorescence and SEM correlative microscopy for nanomanipulation of subcellular structures

Original Articles

- Fluorescence and SEM correlative microscopy for nanomanipulation of subcellular structures
- Fluorescence and SEM correlative microscopy for nanomanipulation of subcellular structures
- LSA 2014年3卷第4期
- Affiliations：
  
  1.Advanced Micro and Nanosystems Laboratory, University of Toronto, Toronto, ON M5S 3G8, Canada
  2.State Key Laboratory of Intelligent Control and Management of Complex Systems, Institute of Automation, Chinese Academy of Sciences, Beijing 100190, China
  3.The Genetics and Genome Biology Program, the Hospital for Sick Children, Toronto, ON M5G 1X8, Canada
- Author bio：
  
  Professor DP Bazett-Jones, the Genetics and Genome Biology Program, the Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canad Email: david.bazett-jones@sickkids.ca
  Professor Y Sun, Advanced Micro and Nanosystems Laboratory, University of Toronto, 5 King's College Road, Toronto, ON M5S 3G8, Canada Email: sun@mie.utoronto.ca
- Funds：
- DOI：10.1038/lsa.2014.105
  中图分类号：
- 纸质出版日期：2014-12，
  
  网络出版日期：2014-11-21，
  
  收稿日期：2014-04-30，
  
  修回日期：2014-09-15，
  
  录用日期：2014-09-16
- Accepted：
Scan QR Code
Fluorescence and SEM correlative microscopy for nanomanipulation of subcellular structures[J]. LSA, 2014,3(4).

Gong, Z. et al. Fluorescence and SEM correlative microscopy for nanomanipulation of subcellular structures. Light: Science & Applications, 3, e224 (2014).
Fluorescence and SEM correlative microscopy for nanomanipulation of subcellular structures[J]. LSA, 2014,3(4). DOI： 10.1038/lsa.2014.105.

Gong, Z. et al. Fluorescence and SEM correlative microscopy for nanomanipulation of subcellular structures. Light: Science & Applications, 3, e224 (2014). DOI： 10.1038/lsa.2014.105.

摘要

Abstract

Nanomanipulation under scanning electron microscopy (SEM) enables direct interactions of a tool with a sample. We recently developed a nanomanipulation technique for the extraction and identification of DNA contained within sub-nuclear locations of a single cell nucleus. In nanomanipulation of sub-cellular structures

key step is to identify targets of interest through correlating fluorescence and SEM images. The DNA extraction task must be conducted with low accelerating voltages resulting in low imaging resolutions. This is imposed by the necessity of preserving the biochemical integrity of the sample. Such poor imaging conditions make the identification of nanometer-sized fiducial marks difficult. This paper presents an affine scale-invariant feature transform (ASIFT) based method for correlating SEM images and fluorescence microscopy images. The performance of the image correlation approach under different noise levels and imaging magnifications was quantitatively evaluated. The optimal mean absolute error (MAE) of correlation results is 68±34 nm under standard conditions. Compared with manual correlation by skilled operators

the automated correlation approach demonstrates a speed that is higher by an order of magnitude. With the SEM-fluorescence image correlation approach

targeted DNA was successfully extracted

via

nanomanipulation under SEM conditions.

关键词

Keywords

correlative microscopyfluorescenceimage correlationnanomanipulationSEMsubcellular structures

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