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Title: Preserving elemental content in adherent mammalian cells for analysis by synchrotron-based x-ray fluorescence microscopy

Abstract

Summary Trace metals play important roles in biological function, and x‐ray fluorescence microscopy (XFM) provides a way to quantitatively image their distribution within cells. The faithfulness of these measurements is dependent on proper sample preparation. Using mouse embryonic fibroblast NIH/3T3 cells as an example, we compare various approaches to the preparation of adherent mammalian cells for XFM imaging under ambient temperature. Direct side‐by‐side comparison shows that plunge‐freezing‐based cryoimmobilization provides more faithful preservation than conventional chemical fixation for most biologically important elements including P, S, Cl, K, Fe, Cu, Zn and possibly Ca in adherent mammalian cells. Although cells rinsed with fresh media had a great deal of extracellular background signal for Cl and Ca, this approach maintained cells at the best possible physiological status before rapid freezing and it does not interfere with XFM analysis of other elements. If chemical fixation has to be chosen, the combination of 3% paraformaldehyde and 1.5 % glutaraldehyde preserves S, Fe, Cu and Zn better than either fixative alone. When chemically fixed cells were subjected to a variety of dehydration processes, air drying was proved to be more suitable than other drying methods such as graded ethanol dehydration and freeze drying. This first detailedmore » comparison for x‐ray fluorescence microscopy shows how detailed quantitative conclusions can be affected by the choice of cell preparation method.« less

Authors:
 [1];  [2];  [3];  [3];  [3];  [3];  [3];  [3];  [2];  [4]
+ Show Author Affiliations
  1. Department of Physics & Astronomy, Weinberg College of Arts and Sciences, Evanston Illinois U.S.A
  2. Department of Radiation Oncology, Northwestern University, Chicago Illinois U.S.A
  3. Advanced Photon Source, Argonne National Laboratory, Argonne Illinois U.S.A
  4. Department of Physics & Astronomy, Weinberg College of Arts and Sciences, Evanston Illinois U.S.A, Advanced Photon Source, Argonne National Laboratory, Argonne Illinois U.S.A
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH)
OSTI Identifier:
1336508
Alternate Identifier(s):
OSTI ID: 1390261; OSTI ID: 1464537
Grant/Contract Number:  
AC02-06CH11357; R01 GM104530; DE‐AC02‐06CH11357
Resource Type:
Published Article
Journal Name:
Journal of Microscopy
Additional Journal Information:
Journal Name: Journal of Microscopy Journal Volume: 265 Journal Issue: 1; Journal ID: ISSN 0022-2720
Publisher:
Wiley
Country of Publication:
United Kingdom
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; Biological; Cryomicroscopy; X-ray microscopy; specimen preparation; x-ray microanalysis

Citation Formats

JIN, QIAOLING, PAUNESKU, TATJANA, LAI, BARRY, GLEBER, SOPHIE-CHARLOTTE, CHEN, SI, FINNEY, LYDIA, VINE, DAVID, VOGT, STEFAN, WOLOSCHAK, GAYLE, and JACOBSEN, CHRIS. Preserving elemental content in adherent mammalian cells for analysis by synchrotron-based x-ray fluorescence microscopy. United Kingdom: N. p., 2016. Web. doi:10.1111/jmi.12466.
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JIN, QIAOLING, PAUNESKU, TATJANA, LAI, BARRY, GLEBER, SOPHIE-CHARLOTTE, CHEN, SI, FINNEY, LYDIA, VINE, DAVID, VOGT, STEFAN, WOLOSCHAK, GAYLE, & JACOBSEN, CHRIS. Preserving elemental content in adherent mammalian cells for analysis by synchrotron-based x-ray fluorescence microscopy. United Kingdom. https://doi.org/10.1111/jmi.12466
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JIN, QIAOLING, PAUNESKU, TATJANA, LAI, BARRY, GLEBER, SOPHIE-CHARLOTTE, CHEN, SI, FINNEY, LYDIA, VINE, DAVID, VOGT, STEFAN, WOLOSCHAK, GAYLE, and JACOBSEN, CHRIS. Wed . "Preserving elemental content in adherent mammalian cells for analysis by synchrotron-based x-ray fluorescence microscopy". United Kingdom. https://doi.org/10.1111/jmi.12466.
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@article{osti_1336508,
title = {Preserving elemental content in adherent mammalian cells for analysis by synchrotron-based x-ray fluorescence microscopy},
author = {JIN, QIAOLING and PAUNESKU, TATJANA and LAI, BARRY and GLEBER, SOPHIE-CHARLOTTE and CHEN, SI and FINNEY, LYDIA and VINE, DAVID and VOGT, STEFAN and WOLOSCHAK, GAYLE and JACOBSEN, CHRIS},
abstractNote = {Summary Trace metals play important roles in biological function, and x‐ray fluorescence microscopy (XFM) provides a way to quantitatively image their distribution within cells. The faithfulness of these measurements is dependent on proper sample preparation. Using mouse embryonic fibroblast NIH/3T3 cells as an example, we compare various approaches to the preparation of adherent mammalian cells for XFM imaging under ambient temperature. Direct side‐by‐side comparison shows that plunge‐freezing‐based cryoimmobilization provides more faithful preservation than conventional chemical fixation for most biologically important elements including P, S, Cl, K, Fe, Cu, Zn and possibly Ca in adherent mammalian cells. Although cells rinsed with fresh media had a great deal of extracellular background signal for Cl and Ca, this approach maintained cells at the best possible physiological status before rapid freezing and it does not interfere with XFM analysis of other elements. If chemical fixation has to be chosen, the combination of 3% paraformaldehyde and 1.5 % glutaraldehyde preserves S, Fe, Cu and Zn better than either fixative alone. When chemically fixed cells were subjected to a variety of dehydration processes, air drying was proved to be more suitable than other drying methods such as graded ethanol dehydration and freeze drying. This first detailed comparison for x‐ray fluorescence microscopy shows how detailed quantitative conclusions can be affected by the choice of cell preparation method.},
doi = {10.1111/jmi.12466},
journal = {Journal of Microscopy},
number = 1,
volume = 265,
place = {United Kingdom},
year = {Wed Aug 31 00:00:00 EDT 2016},
month = {Wed Aug 31 00:00:00 EDT 2016}
}
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Works referenced in this record:

Imaging and speciation of trace elements in biological environment
journal, November 2006

Characterization of Intra‐Cellular Ionic Concentrations of Monocytes in Contact with Bioactive Glasses and Hydroxyapatite Particles. A TEM Cryo‐X‐ray Analysis of Diffusible Ions
journal, December 2004

  • Jallot, E.; Laquerriere, P.; Grandjean‐Laquerriere, A.
  • Instrumentation Science & Technology, Vol. 32, Issue 5
  • DOI: 10.1081/CI-200029800

Imaging trace element distributions in single organelles and subcellular features
journal, February 2016

  • Kashiv, Yoav; Austin, Jotham R.; Lai, Barry
  • Scientific Reports, Vol. 6, Issue 1
  • DOI: 10.1038/srep21437

Trends in hard X-ray fluorescence mapping: environmental applications in the age of fast detectors
journal, March 2011

  • Lombi, E.; de Jonge, M. D.; Donner, E.
  • Analytical and Bioanalytical Chemistry, Vol. 400, Issue 6
  • DOI: 10.1007/s00216-011-4829-2

X-ray fluorescence microprobe imaging in biology and medicine
journal, January 2006

  • Paunesku, Tatjana; Vogt, Stefan; Maser, Jörg
  • Journal of Cellular Biochemistry, Vol. 99, Issue 6
  • DOI: 10.1002/jcb.21047

Metal imaging in neurodegenerative diseases
journal, January 2012

  • Bourassa, Megan W.; Miller, Lisa M.
  • Metallomics, Vol. 4, Issue 8
  • DOI: 10.1039/c2mt20052j

Subcellular redistribution and mitotic inheritance of transition metals in proliferating mouse fibroblast cells
journal, January 2013

  • McRae, Reagan; Lai, Barry; Fahrni, Christoph J.
  • Metallomics, Vol. 5, Issue 1
  • DOI: 10.1039/C2MT20176C

Subcellular Speciation Analysis of Trace Element Oxidation States Using Synchrotron Radiation Micro-X-ray Absorption Near-Edge Structure
journal, October 2007

  • Bacquart, Thomas; Devès, Guillaume; Carmona, Asunción
  • Analytical Chemistry, Vol. 79, Issue 19
  • DOI: 10.1021/ac0711135

Unsupervised cell identification on multidimensional X-ray fluorescence datasets
journal, March 2014

In Situ Imaging of Metals in Cells and Tissues
journal, October 2009

  • McRae, Reagan; Bagchi, Pritha; Sumalekshmy, S.
  • Chemical Reviews, Vol. 109, Issue 10
  • DOI: 10.1021/cr900223a

Chemical alterations to murine brain tissue induced by formalin fixation: implications for biospectroscopic imaging and mapping studies of disease pathogenesis
journal, January 2011

  • Hackett, Mark J.; McQuillan, James A.; El-Assaad, Fatima
  • The Analyst, Vol. 136, Issue 14
  • DOI: 10.1039/c0an00269k

Atrial-specific granules in situ have high calcium content, are acidic, and maintain anion gradients.
journal, August 1988

  • Somlyo, A. V.; Broderick, R.; Shuman, H.
  • Proceedings of the National Academy of Sciences, Vol. 85, Issue 16
  • DOI: 10.1073/pnas.85.16.6222

Preparation of biological cryosections for analytical electron microscopy
journal, January 1982

Electron microscopy of high pressure frozen samples: bridging the gap between cellular ultrastructure and atomic resolution
journal, September 2008

  • Studer, Daniel; Humbel, Bruno M.; Chiquet, Matthias
  • Histochemistry and Cell Biology, Vol. 130, Issue 5
  • DOI: 10.1007/s00418-008-0500-1

Elemental and Redox Analysis of Single Bacterial Cells by X-ray Microbeam Analysis
journal, October 2004

Vitrification depth can be increased more than 10-fold by high-pressure freezing
journal, October 1993

Preparation of tissue samples for X-ray fluorescence microscopy
journal, December 2005

  • Chwiej, Joanna; Szczerbowska-Boruchowska, Magdalena; Lankosz, Marek
  • Spectrochimica Acta Part B: Atomic Spectroscopy, Vol. 60, Issue 12
  • DOI: 10.1016/j.sab.2005.10.002

Opportunities in multidimensional trace metal imaging: taking copper-associated disease research to the next level
journal, October 2012

Time-dependent uptake, distribution and biotransformation of chromium(VI) in individual and bulk human lung cells: application of synchrotron radiation techniques
journal, February 2005

  • Harris, Hugh H.; Levina, Aviva; Dillon, Carolyn T.
  • JBIC Journal of Biological Inorganic Chemistry, Vol. 10, Issue 2
  • DOI: 10.1007/s00775-004-0617-1

Nanocarriers Enhance Doxorubicin Uptake in Drug-Resistant Ovarian Cancer Cells
journal, December 2011

Imaging methods for elemental, chemical, molecular, and morphological analyses of single cells
journal, March 2010

Cryofixation Without Pretreatment at Ambient Pressure
book, January 1987

Electron microscopy of frozen water and aqueous solutions
journal, December 1982

MAPS : A set of software tools for analysis and visualization of 3D X-ray fluorescence data sets
journal, March 2003

Hard X-ray microprobe studies of chromium(VI)-treated V79 Chinese hamster lung cells: intracellular mapping of the biotransformation products of a chromium carcinogen
journal, February 2002

  • Dillon, Carolyn T.; Lay, Peter A.; Kennedy, Brendan J.
  • JBIC Journal of Biological Inorganic Chemistry, Vol. 7, Issue 6
  • DOI: 10.1007/s00775-002-0343-5

Quantifying Trace Elements in Individual Aquatic Protist Cells with a Synchrotron X-ray Fluorescence Microprobe
journal, August 2003

  • Twining, Benjamin S.; Baines, Stephen B.; Fisher, Nicholas S.
  • Analytical Chemistry, Vol. 75, Issue 15
  • DOI: 10.1021/ac034227z

Calcium content of mitochondria and endoplasmic reticulum in liver frozen rapidly in vivo
journal, April 1985

  • Somlyo, A. P.; Bond, Meredith; Somlyo, Avril V.
  • Nature, Vol. 314, Issue 6012
  • DOI: 10.1038/314622a0

Combined use of hard X-ray phase contrast imaging and X-ray fluorescence microscopy for sub-cellular metal quantification
journal, February 2012

  • Kosior, Ewelina; Bohic, Sylvain; Suhonen, Heikki
  • Journal of Structural Biology, Vol. 177, Issue 2
  • DOI: 10.1016/j.jsb.2011.12.005

Elemental mapping of frozen-hydrated cells with cryo-scanning X-ray fluorescence microscopy
journal, May 2010

  • Matsuyama, S.; Shimura, M.; Fujii, M.
  • X-Ray Spectrometry, Vol. 39, Issue 4
  • DOI: 10.1002/xrs.1256

Rapid and Accurate Analysis of an X-Ray Fluorescence Microscopy Data Set through Gaussian Mixture-Based Soft Clustering Methods
journal, August 2013

  • Ward, Jesse; Marvin, Rebecca; O'Halloran, Thomas
  • Microscopy and Microanalysis, Vol. 19, Issue 5
  • DOI: 10.1017/S1431927613012737

Uptake, Distribution, and Speciation of Selenoamino Acids by Human Cancer Cells: X-ray Absorption and Fluorescence Methods
journal, March 2011

  • Weekley, Claire M.; Aitken, Jade B.; Vogt, Stefan
  • Biochemistry, Vol. 50, Issue 10
  • DOI: 10.1021/bi101678a

Quantitative imaging of free and total intracellular calcium in cultured cells.
journal, March 1989

  • Chandra, S.; Gross, D.; Ling, Y. C.
  • Proceedings of the National Academy of Sciences, Vol. 86, Issue 6
  • DOI: 10.1073/pnas.86.6.1870

Activity-dependent accumulation of calcium in Purkinje cell dendritic spines.
journal, March 1988

  • Andrews, S. B.; Leapman, R. D.; Landis, D. M.
  • Proceedings of the National Academy of Sciences, Vol. 85, Issue 5
  • DOI: 10.1073/pnas.85.5.1682

Distribution of calcium and potassium in presynaptic nerve terminals from cerebellar cortex.
journal, March 1987

  • Andrews, S. B.; Leapman, R. D.; Landis, D. M.
  • Proceedings of the National Academy of Sciences, Vol. 84, Issue 6
  • DOI: 10.1073/pnas.84.6.1713

Quantitative electron probe microanalysis of biological thin sections: Methods and validity
journal, January 1976

Correlative microXRF and optical immunofluorescence microscopy of adherent cells labeled with ultrasmall gold particles
journal, July 2006

Mass spectrometric characterization of elements and molecules in cell cultures and tissues
journal, July 2006

The Bionanoprobe: hard X-ray fluorescence nanoprobe with cryogenic capabilities
journal, December 2013

Silicon nitride as a versatile growth substrate for microspectroscopic imaging and mapping of individual cells
journal, January 2010

  • Carter, Elizabeth A.; Rayner, Benjamin S.; McLeod, Andrew I.
  • Molecular BioSystems, Vol. 6, Issue 7
  • DOI: 10.1039/c001499k

Sample preparation of animal tissues and cell cultures for secondary ion mass spectrometry (SIMS) microscopy
journal, January 1992

Diffusible ion localization by ion microscopy: a comparison of chemically prepared and fast-frozen, freeze-dried, unfixed liver sections
journal, April 1980

Iron Storage within Dopamine Neurovesicles Revealed by Chemical Nano-Imaging
journal, September 2007

Biological applications of X-ray fluorescence microscopy: exploring the subcellular topography and speciation of transition metals
journal, April 2007

Correlative VIS-fluorescence and soft X-ray cryo-microscopy/tomography of adherent cells
journal, February 2012

  • Hagen, Christoph; Guttmann, Peter; Klupp, Barbara
  • Journal of Structural Biology, Vol. 177, Issue 2
  • DOI: 10.1016/j.jsb.2011.12.012

X-ray fluorescence spectrometry, an analytical tool in neurochemical research
journal, January 2008

  • Szczerbowska-Boruchowska, Magdalena
  • X-Ray Spectrometry, Vol. 37, Issue 1
  • DOI: 10.1002/xrs.1007

Melanosomal sequestration of cytotoxic drugs contributes to the intractability of malignant melanomas
journal, June 2006

  • Chen, K. G.; Valencia, J. C.; Lai, B.
  • Proceedings of the National Academy of Sciences, Vol. 103, Issue 26
  • DOI: 10.1073/pnas.0600213103

Evaluation of sample preparation methods for single cell quantitative elemental imaging using proton or synchrotron radiation focused beams
journal, January 2015

  • Perrin, Laura; Carmona, Asuncion; Roudeau, Stéphane
  • Journal of Analytical Atomic Spectrometry, Vol. 30, Issue 12
  • DOI: 10.1039/C5JA00303B

Trace element mapping of a single cell using a hard x-ray nanobeam focused by a Kirkpatrick-Baez mirror system
journal, March 2009

  • Matsuyama, S.; Shimura, M.; Mimura, H.
  • X-Ray Spectrometry, Vol. 38, Issue 2
  • DOI: 10.1002/xrs.1123

Applications of synchrotron μ-XRF to study the distribution of biologically important elements in different environmental matrices: A review
journal, November 2012

  • Majumdar, Sanghamitra; Peralta-Videa, Jose R.; Castillo-Michel, Hiram
  • Analytica Chimica Acta, Vol. 755
  • DOI: 10.1016/j.aca.2012.09.050

Biology of TiO2–oligonucleotide nanocomposites
journal, April 2003

  • Paunesku, Tatjana; Rajh, Tijana; Wiederrecht, Gary
  • Nature Materials, Vol. 2, Issue 5
  • DOI: 10.1038/nmat875

Bio-metals imaging and speciation in cells using proton and synchrotron radiation X-ray microspectroscopy
journal, June 2009

  • Ortega, Richard; Devès, Guillaume; Carmona, Asunción
  • Journal of The Royal Society Interface, Vol. 6, Issue suppl_5
  • DOI: 10.1098/rsif.2009.0166.focus

Advantages of fast-freeze fixation followed by freeze-substitution for the preservation of cell integrity
journal, August 1991

Cryofixation methods for ion localization in cells by electron probe microanalysis: a review
journal, February 1991

X-ray microanalysis of cultured mammalian cells
book, April 1993

Cellular distribution and degradation of cobalt ferrite nanoparticles in Balb/3T3 mouse fibroblasts
journal, November 2011

The localisation and micro-mapping of copper and other trace elements in breast tumours using a synchrotron micro-XRF system
journal, February 2007

Synchrotron-based X-ray fluorescence imaging of human cells labeled with CdSe quantum dots
journal, May 2009

  • Corezzi, Silvia; Urbanelli, Lorena; Cloetens, Peter
  • Analytical Biochemistry, Vol. 388, Issue 1
  • DOI: 10.1016/j.ab.2009.01.044

Fixation and Drying Protocols for the Preparation of Cell Samples for Time-of-Flight Secondary Ion Mass Spectrometry Analysis
journal, September 2009

  • Malm, Jakob; Giannaras, Dimitrios; Riehle, Mathis O.
  • Analytical Chemistry, Vol. 81, Issue 17
  • DOI: 10.1021/ac900636v

Fluxes in “Free” and Total Zinc Are Essential for Progression of Intraerythrocytic Stages of Plasmodium falciparum
journal, June 2012

Calcium measurements with electron probe X-ray and electron energy loss analysis.
journal, March 1990

  • LeFurgey, A.; Ingram, P.
  • Environmental Health Perspectives, Vol. 84
  • DOI: 10.1289/ehp.908457

Increased brain iron coincides with early plaque formation in a mouse model of Alzheimer's disease
journal, March 2011

Nuclear microscopy in Alzheimer’s disease
journal, April 1999

  • Makjanic, Jagoda; Watt, Frank
  • Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms, Vol. 150, Issue 1-4
  • DOI: 10.1016/S0168-583X(98)01033-7

Study of Cysteinyl Leukotriene-1 Receptor in Rat Renal Ischemia-Reperfusion Injury
journal, September 2008

Imaging of the intracellular topography of copper with a fluorescent sensor and by synchrotron x-ray fluorescence microscopy
journal, August 2005

  • Yang, L.; McRae, R.; Henary, M. M.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 32
  • DOI: 10.1073/pnas.0406547102

Synchrotron hard x-ray microprobe: Fluorescence imaging of single cells
journal, May 2001

  • Bohic, S.; Simionovici, A.; Snigirev, A.
  • Applied Physics Letters, Vol. 78, Issue 22
  • DOI: 10.1063/1.1366362

High-pressure freezing for immunocytochemistry
journal, December 1998

X-ray fluorescence microscopy reveals large-scale relocalization and extracellular translocation of cellular copper during angiogenesis
journal, February 2007

  • Finney, L.; Mandava, S.; Ursos, L.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 7
  • DOI: 10.1073/pnas.0607238104

X-ray fluorescence imaging reveals subcellular biometal disturbances in a childhood neurodegenerative disorder
journal, January 2014

  • Grubman, A.; James, S. A.; James, J.
  • Chem. Sci., Vol. 5, Issue 6
  • DOI: 10.1039/C4SC00316K

Loss of Pluripotency in Human Embryonic Stem Cells Directly Correlates with an Increase in Nuclear Zinc
journal, August 2010

Advances in ultrarapid freezing for the preservation of cellular ultrastructure
journal, January 1986

  • Gilkey, John C.; Staehelin, L. Andrew
  • Journal of Electron Microscopy Technique, Vol. 3, Issue 2, p. 177-210
  • DOI: 10.1002/jemt.1060030206

Quantitative digital X-ray imaging using frozen hydrated and frozen dried tissue sections
journal, May 1987

Application of scanning electron microscopy to x-ray analysis of frozen- hydrated sections. I. Specimen handling techniques
journal, February 1981

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