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Cell Imaging Techniques pp 85–95Cite as

Multifluorescence Confocal Microscopy: Application for a Quantitative Analysis of Hemostatic Proteins in Human Venous Valves

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 931))

Abstract

Confocal laser scanning microscopy is commonly used to visualize and quantify protein expression. Visualization of the expression of multiple proteins in the same region via multifluorescence allows for the analysis of differential protein expression. The defining step of multifluorescence labeling is the selection of primary antibodies from different host species. In addition, species-appropriate secondary antibodies must also be conjugated to different fluorophores so that each protein can be visualized in separate channels. Quantitative analysis of proteins labeled via multifluorescence can be used to compare relative changes in protein expression. Multifluoresecence labeling and analysis of fluorescence intensity within and among human venous specimens, for example, allowed us to determine that the anticoagulant phenotype of the venous valve is defined not by increased anticoagulant expression, but instead by significantly decreased procoagulant protein expression (Blood 114:1276–1279, 2009 and Histochem Cell Biol 135:141–152, 2011).

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References

  1. Brooks EG, Trotman WE, Wadsworth MP, Taatjes DJ, Evans MF, Ittleman FP, Callas PW, Esmon CT, Bovill EG (2009) Valves of the deep venous system: an overlooked risk factor. Blood 114:1276–1279

    Article  PubMed  CAS  Google Scholar 

  2. Trotman WE, Taatjes DJ, Callas PW, Bovill EG (2011) The endothelial microenvironment in the venous valvular sinus: thromboresistance trends and inter-individual variation. Histochem Cell Biol 135:141–152

    Article  PubMed  CAS  Google Scholar 

  3. Lichtman JW, Conchello J-A (2005) Fluorescence microscopy. Nat Methods 2:910–919

    Article  PubMed  CAS  Google Scholar 

  4. Conchello J-A, Lichtman JW (2005) Optical sectioning microscopy. Nat Methods 2:920–931

    Article  PubMed  CAS  Google Scholar 

  5. Amos WB, White JG (2003) How the confocal laser scanning microscope entered biological research. Biol Cell 95:335–342

    Article  PubMed  CAS  Google Scholar 

  6. Stern M, Taatjes DJ, Mossman BT (2005) Multifluorescence labeling techniques and confocal laser scanning microscopy on lung tissue. Methods Mol Biol 319:67–76

    Article  Google Scholar 

  7. Sedgewick J (2007) Guidelines for a specific type of images. Scientific imaging with Photoshop: methods, measurements and output. New Riders, Berkeley, pp 44–74

    Google Scholar 

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Acknowledgments

The project described in this chapter was supported by Award Number 1S10RR019246 from the National Center for Research Resources (to DJT) for purchase of the Zeiss 510 META confocal scanning laser microscope.

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Authors and Affiliations

  1. Department of Pathology, College of Medicine, University of Vermont, Burlington, VT, USA

    Winifred E. Trotman & Edwin G. Bovill

  2. Department of Pathology and Microscopy Imaging Center, College of Medicine, University of Vermont, Burlington, VT, USA

    Douglas J. Taatjes

Authors
  1. Winifred E. Trotman
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  2. Douglas J. Taatjes
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  3. Edwin G. Bovill
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Corresponding author

Correspondence to Douglas J. Taatjes .

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Editors and Affiliations

  1. College of Medicine, Professor, Department of Pathology, University of Vermont, Beaumont Ave 89, Burlington, 05405-1742, Vermont, USA

    Douglas J. Taatjes

  2. , Abt. Zell- und Molekularpathologie, Universität Zürich, Schmelzbergstr. 12, Zürich, 8091, Switzerland

    Jürgen Roth

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Trotman, W.E., Taatjes, D.J., Bovill, E.G. (2012). Multifluorescence Confocal Microscopy: Application for a Quantitative Analysis of Hemostatic Proteins in Human Venous Valves. In: Taatjes, D., Roth, J. (eds) Cell Imaging Techniques. Methods in Molecular Biology, vol 931. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-056-4_4

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  • DOI: https://doi.org/10.1007/978-1-62703-056-4_4

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-055-7

  • Online ISBN: 978-1-62703-056-4

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