Development of cell staining technique for X-ray microscopy

P. Y. Tseng; Y. T. Shih; C. J. Liu; T. Hsu; C. C. Chien; W. H. Leng; K. S. Liang; G. C. Yin; F. R. Chen; J. H. Je; G. Margaritondo; Y. Hwu

doi:10.1063/1.2436459

Development of cell staining technique for X-ray microscopy

P. Y. Tseng
, Y. T. Shih
, C. J. Liu
, T. Hsu
, C. C. Chien
, W. H. Leng
, K. S. Liang
, G. C. Yin
, F. R. Chen
, J. H. Je
, G. Margaritondo
, Y. Hwu

Research output: Chapters, Conference Papers, Creative and Literary Works › RGC 32 - Refereed conference paper (with host publication) › peer-review

2 Citations (Scopus)

Abstract

We report a technique for detection of sub-cellular organelles and proteins with hard x-ray microscopy. Several metals were used for enhancing contrast for x-ray microscopy. Osmium tetroxide provides an excellent stain for lipid and can delineate cell membrane. Uranyl acetate has high affinity for nucleotide and can stain nucleus. Immunolocalization of specific proteins and sub-cellular organelles was achieved by 3'3 diaminobenzidine (DAB) with nickel enhancement and nanogold-conjugated secondary antibody with silver enhancement. The x-rays emitted from synchrotron source was monochromatized by double crystal monochromator, the photon energy was fixed at 8 keV to optimize the focusing efficiency of the zone plates. The estimated resolution is about 60 nm. When compared with visible light and conventional confocal microscopy, the X-ray microscopy provides a superior resolution to both conventional optical microscopes.

Original language	English
Title of host publication	SYNCHROTRON RADIATION INSTRUMENTATION
Subtitle of host publication	Ninth International Conference on Synchrotron Radiation Instrumentation
Publisher	AIP Publishing
Pages	1964-1967
ISBN (Print)	9780735403734
DOIs	https://doi.org/10.1063/1.2436459
Publication status	Published - 19 Jan 2007
Externally published	Yes
Event	9th International Conference on Synchrotron Radiation Instrumentation - Daegu, Korea, Republic of Duration: 28 May 2006 → 2 Jun 2006 https://cds.cern.ch/record/896538

Publication series

Name	AIP Conference Proceedings
Number	1
Volume	879
ISSN (Print)	0094-243X
ISSN (Electronic)	1551-7616

Conference

Conference	9th International Conference on Synchrotron Radiation Instrumentation
Place	Korea, Republic of
City	Daegu
Period	28/05/06 → 2/06/06
Internet address	https://cds.cern.ch/record/896538

Research Keywords

Cell staining
Hard x-ray
Protein localization
X-ray microscopy

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10.1063/1.2436459

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Tseng, P. Y., Shih, Y. T., Liu, C. J., Hsu, T., Chien, C. C., Leng, W. H., Liang, K. S., Yin, G. C., Chen, F. R., Je, J. H., Margaritondo, G., & Hwu, Y. (2007). Development of cell staining technique for X-ray microscopy. In SYNCHROTRON RADIATION INSTRUMENTATION: Ninth International Conference on Synchrotron Radiation Instrumentation (pp. 1964-1967). (AIP Conference Proceedings; Vol. 879, No. 1). AIP Publishing. https://doi.org/10.1063/1.2436459

@inproceedings{c3c3f61895a148a48a6a098cc67891e3,

title = "Development of cell staining technique for X-ray microscopy",

abstract = "We report a technique for detection of sub-cellular organelles and proteins with hard x-ray microscopy. Several metals were used for enhancing contrast for x-ray microscopy. Osmium tetroxide provides an excellent stain for lipid and can delineate cell membrane. Uranyl acetate has high affinity for nucleotide and can stain nucleus. Immunolocalization of specific proteins and sub-cellular organelles was achieved by 3'3 diaminobenzidine (DAB) with nickel enhancement and nanogold-conjugated secondary antibody with silver enhancement. The x-rays emitted from synchrotron source was monochromatized by double crystal monochromator, the photon energy was fixed at 8 keV to optimize the focusing efficiency of the zone plates. The estimated resolution is about 60 nm. When compared with visible light and conventional confocal microscopy, the X-ray microscopy provides a superior resolution to both conventional optical microscopes. ",

keywords = "Cell staining, Hard x-ray, Protein localization, X-ray microscopy",

author = "Tseng, \{P. Y.\} and Shih, \{Y. T.\} and Liu, \{C. J.\} and T. Hsu and Chien, \{C. C.\} and Leng, \{W. H.\} and Liang, \{K. S.\} and Yin, \{G. C.\} and Chen, \{F. R.\} and Je, \{J. H.\} and G. Margaritondo and Y. Hwu",

year = "2007",

month = jan,

day = "19",

doi = "10.1063/1.2436459",

language = "English",

isbn = "9780735403734",

series = "AIP Conference Proceedings",

publisher = "AIP Publishing",

number = "1",

pages = "1964--1967",

booktitle = "SYNCHROTRON RADIATION INSTRUMENTATION",

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note = "9th International Conference on Synchrotron Radiation Instrumentation ; Conference date: 28-05-2006 Through 02-06-2006",

url = "https://cds.cern.ch/record/896538",

}

Tseng, PY, Shih, YT, Liu, CJ, Hsu, T, Chien, CC, Leng, WH, Liang, KS, Yin, GC, Chen, FR, Je, JH, Margaritondo, G & Hwu, Y 2007, Development of cell staining technique for X-ray microscopy. in SYNCHROTRON RADIATION INSTRUMENTATION: Ninth International Conference on Synchrotron Radiation Instrumentation. AIP Conference Proceedings, no. 1, vol. 879, AIP Publishing, pp. 1964-1967, 9th International Conference on Synchrotron Radiation Instrumentation, Daegu, Korea, Republic of, 28/05/06. https://doi.org/10.1063/1.2436459

Development of cell staining technique for X-ray microscopy. / Tseng, P. Y.; Shih, Y. T.; Liu, C. J. et al.
SYNCHROTRON RADIATION INSTRUMENTATION: Ninth International Conference on Synchrotron Radiation Instrumentation. AIP Publishing, 2007. p. 1964-1967 (AIP Conference Proceedings; Vol. 879, No. 1).

Research output: Chapters, Conference Papers, Creative and Literary Works › RGC 32 - Refereed conference paper (with host publication) › peer-review

TY - GEN

T1 - Development of cell staining technique for X-ray microscopy

AU - Tseng, P. Y.

AU - Shih, Y. T.

AU - Liu, C. J.

AU - Hsu, T.

AU - Chien, C. C.

AU - Leng, W. H.

AU - Liang, K. S.

AU - Yin, G. C.

AU - Chen, F. R.

AU - Je, J. H.

AU - Margaritondo, G.

AU - Hwu, Y.

PY - 2007/1/19

Y1 - 2007/1/19

N2 - We report a technique for detection of sub-cellular organelles and proteins with hard x-ray microscopy. Several metals were used for enhancing contrast for x-ray microscopy. Osmium tetroxide provides an excellent stain for lipid and can delineate cell membrane. Uranyl acetate has high affinity for nucleotide and can stain nucleus. Immunolocalization of specific proteins and sub-cellular organelles was achieved by 3'3 diaminobenzidine (DAB) with nickel enhancement and nanogold-conjugated secondary antibody with silver enhancement. The x-rays emitted from synchrotron source was monochromatized by double crystal monochromator, the photon energy was fixed at 8 keV to optimize the focusing efficiency of the zone plates. The estimated resolution is about 60 nm. When compared with visible light and conventional confocal microscopy, the X-ray microscopy provides a superior resolution to both conventional optical microscopes.

AB - We report a technique for detection of sub-cellular organelles and proteins with hard x-ray microscopy. Several metals were used for enhancing contrast for x-ray microscopy. Osmium tetroxide provides an excellent stain for lipid and can delineate cell membrane. Uranyl acetate has high affinity for nucleotide and can stain nucleus. Immunolocalization of specific proteins and sub-cellular organelles was achieved by 3'3 diaminobenzidine (DAB) with nickel enhancement and nanogold-conjugated secondary antibody with silver enhancement. The x-rays emitted from synchrotron source was monochromatized by double crystal monochromator, the photon energy was fixed at 8 keV to optimize the focusing efficiency of the zone plates. The estimated resolution is about 60 nm. When compared with visible light and conventional confocal microscopy, the X-ray microscopy provides a superior resolution to both conventional optical microscopes.

KW - Cell staining

KW - Hard x-ray

KW - Protein localization

KW - X-ray microscopy

UR - https://www.scopus.com/pages/publications/33947422078

UR - https://www.scopus.com/record/pubmetrics.uri?eid=2-s2.0-33947422078&origin=recordpage

U2 - 10.1063/1.2436459

DO - 10.1063/1.2436459

M3 - RGC 32 - Refereed conference paper (with host publication)

SN - 9780735403734

T3 - AIP Conference Proceedings

SP - 1964

EP - 1967

BT - SYNCHROTRON RADIATION INSTRUMENTATION

PB - AIP Publishing

T2 - 9th International Conference on Synchrotron Radiation Instrumentation

Y2 - 28 May 2006 through 2 June 2006

ER -

Development of cell staining technique for X-ray microscopy

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