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Title: Micro-CT scouting for transmission electron microscopy of human tissue specimens

Abstract

Transmission electron microscopy (TEM) provides sub-nanometre-scale details in volumetric samples. Samples such as pathology tissue specimens are often stained with a metal element to enhance contrast, which makes them opaque to optical microscopes. As a result, it can be a lengthy procedure to find the region of interest inside a sample through sectioning. Here, we describe micro-CT scouting for TEM that allows noninvasive identification of regions of interest within a block sample to guide the sectioning step. In a tissue pathology study, a bench-top micro-CT scanner with 10 m resolution was used to determine the location of patches of the mucous membrane in osmium-stained human nasal scraping samples. Furthermore, once the regions of interest were located, the sample block was sectioned to expose that location, followed by ultra-thin sectioning and TEM to inspect the internal structure of the cilia of the membrane epithelial cells with nanometre resolution. This method substantially reduced the time and labour of the search process from typically 20 sections for light microscopy to three sections with no added sample preparation. Lay description Electron microscopy provides very high levels of detail in a small area, and thus the question of where to look in an opaque sample,more » such as a stained tissue specimen, needs to be answered by sectioning the sample in small steps and examining the sections under a light microscope, until the region of interest is found. The search process can be lengthy and labor intensive, especially for a study involving a large number of samples. Small areas of interest can be missed in the process if not enough regions are examined. We also describe a method to directly locate the region of interest within a whole sample using micro-CT imaging, bypassing the need of blindly sectioning. Micro-CT enables locating the region within 3D space; this information provides a guide for sectioning the sample to expose that precise location for high resolution electron microscopy imaging. In a human tissue specimen study, this method considerably reduced the time and labor of the search process.« less

Authors:
 [1];  [1];  [2];  [1];  [1];  [1];  [1];  [1];  [1];  [3];  [1];  [1]
+ Show Author Affiliations
  1. National Inst. of Health (NIH), Bethesda, MD (United States). National Heart Lung and Blood Inst.
  2. Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
  3. National Inst. of Health (NIH), Bethesda, MD (United States). National Inst. of Neurological Disorders and Stroke
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1393471
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Microscopy
Additional Journal Information:
Journal Volume: 263; Journal Issue: 1; Journal ID: ISSN 0022-2720
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; micro-CT scouting; bench-top micro-CT scanner; three-dimensional visualization; transmission electron microscopy

Citation Formats

Morales, A. G., Stempinski, E. S., XIAO, X., Patel, A., Panna, A., Oliver, K. N., McShane, P. J., Robinson, C., George, A. J., Donahue, D. R., Chen, P., and Wen, H. Micro-CT scouting for transmission electron microscopy of human tissue specimens. United States: N. p., 2016. Web. doi:10.1111/jmi.12385.
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Morales, A. G., Stempinski, E. S., XIAO, X., Patel, A., Panna, A., Oliver, K. N., McShane, P. J., Robinson, C., George, A. J., Donahue, D. R., Chen, P., & Wen, H. Micro-CT scouting for transmission electron microscopy of human tissue specimens. United States. https://doi.org/10.1111/jmi.12385
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Morales, A. G., Stempinski, E. S., XIAO, X., Patel, A., Panna, A., Oliver, K. N., McShane, P. J., Robinson, C., George, A. J., Donahue, D. R., Chen, P., and Wen, H. Mon . "Micro-CT scouting for transmission electron microscopy of human tissue specimens". United States. https://doi.org/10.1111/jmi.12385. https://www.osti.gov/servlets/purl/1393471.
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@article{osti_1393471,
title = {Micro-CT scouting for transmission electron microscopy of human tissue specimens},
author = {Morales, A. G. and Stempinski, E. S. and XIAO, X. and Patel, A. and Panna, A. and Oliver, K. N. and McShane, P. J. and Robinson, C. and George, A. J. and Donahue, D. R. and Chen, P. and Wen, H.},
abstractNote = {Transmission electron microscopy (TEM) provides sub-nanometre-scale details in volumetric samples. Samples such as pathology tissue specimens are often stained with a metal element to enhance contrast, which makes them opaque to optical microscopes. As a result, it can be a lengthy procedure to find the region of interest inside a sample through sectioning. Here, we describe micro-CT scouting for TEM that allows noninvasive identification of regions of interest within a block sample to guide the sectioning step. In a tissue pathology study, a bench-top micro-CT scanner with 10 m resolution was used to determine the location of patches of the mucous membrane in osmium-stained human nasal scraping samples. Furthermore, once the regions of interest were located, the sample block was sectioned to expose that location, followed by ultra-thin sectioning and TEM to inspect the internal structure of the cilia of the membrane epithelial cells with nanometre resolution. This method substantially reduced the time and labour of the search process from typically 20 sections for light microscopy to three sections with no added sample preparation. Lay description Electron microscopy provides very high levels of detail in a small area, and thus the question of where to look in an opaque sample, such as a stained tissue specimen, needs to be answered by sectioning the sample in small steps and examining the sections under a light microscope, until the region of interest is found. The search process can be lengthy and labor intensive, especially for a study involving a large number of samples. Small areas of interest can be missed in the process if not enough regions are examined. We also describe a method to directly locate the region of interest within a whole sample using micro-CT imaging, bypassing the need of blindly sectioning. Micro-CT enables locating the region within 3D space; this information provides a guide for sectioning the sample to expose that precise location for high resolution electron microscopy imaging. In a human tissue specimen study, this method considerably reduced the time and labor of the search process.},
doi = {10.1111/jmi.12385},
journal = {Journal of Microscopy},
number = 1,
volume = 263,
place = {United States},
year = {Mon Feb 08 00:00:00 EST 2016},
month = {Mon Feb 08 00:00:00 EST 2016}
}
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https://doi.org/10.1111/jmi.12385
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