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Confocal Imaging and Three-Dimensional Visualization of Thick Autofluorescent Specimens

  • Angela V. Klaus
  • Valerie Schawaroch
  • Kevin J. Frischmann
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1075)

Abstract

Three-dimensional (3-D) rendering methods (maximum intensity projection, alpha blending, and isosurface rendering) are described for the visualization of thick, autofluorescent, arthropod cuticular structures (e.g., Drosophila melanogaster external genitalic structures) imaged by confocal laser scanning microscopy (CLSM). Additionally, specimen mounting and data collection strategies for thick specimens are described. Axial aberration artifacts are discussed in the context of these methods because of the critical roles they play in the quality of final 3-D images.

Key words

Confocal microscopy Spherical aberration Drosophila Maximum intensity projection Volume rendering Surface rendering Three-dimensional reconstruction Volume visualization Axial aberration 

Notes

Acknowledgments

VS acknowledges the generous support for this work from a National Science Foundation award (DEB0075360), two PSC-CUNY awards (60052-34-35 and 67621-00-36), and a Eugene M. Lang Junior Faculty Research Fellowship. VS also wishes to thank Dean Myrna Chase of the Weisman School of Arts and Sciences of Baruch College for reassigned time.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Angela V. Klaus
    • 1
  • Valerie Schawaroch
    • 2
    • 3
  • Kevin J. Frischmann
    • 4
  1. 1.Department of BiologySeton Hall UniversitySouth OrangeUSA
  2. 2.Department of Natural SciencesBaruch CollegeNew YorkUSA
  3. 3.Division of Invertebrate ZoologyAmerican Museum of Natural HistoryNew YorkUSA
  4. 4.Bitplane Inc.Saint PaulUSA

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