title: Showing their true colors: a practical approach to volume rendering from serial sections.
abstract: BACKGROUND
  In comparison to more modern imaging methods, conventional light microscopy still offers a range of substantial advantages with regard to contrast options, accessible specimen size, and resolution. Currently, tomographic image data in particular is most commonly visualized in three dimensions using volume rendering. To date, this method has only very rarely been applied to image stacks taken from serial sections, whereas surface rendering is still the most prevalent method for presenting such data sets three-dimensionally. The aim of this study was to develop standard protocols for volume rendering of image stacks of serial sections, while retaining the benefits of light microscopy such as resolution and color information.
  
  
  RESULTS
  Here we provide a set of protocols for acquiring high-resolution 3D images of diverse microscopic samples through volume rendering based on serial light microscopical sections using the 3D reconstruction software Amira (Visage Imaging Inc.). We overcome several technical obstacles and show that these renderings are comparable in quality and resolution to 3D visualizations using other methods. This practical approach for visualizing 3D micro-morphology in full color takes advantage of both the sub-micron resolution of light microscopy and the specificity of histological stains, by combining conventional histological sectioning techniques, digital image acquisition, three-dimensional image filtering, and 3D image manipulation and visualization technologies.
  
  
  CONCLUSIONS
  We show that this method can yield "true"-colored high-resolution 3D views of tissues as well as cellular and sub-cellular structures and thus represents a powerful tool for morphological, developmental, and comparative investigations. We conclude that the presented approach fills an important gap in the field of micro-anatomical 3D imaging and visualization methods by combining histological resolution and differentiation of details with 3D rendering of whole tissue samples. We demonstrate the method on selected invertebrate and vertebrate specimens, and propose that reinvestigation of historical serial section material may be regarded as a special benefit.
authors.affiliation: Department of Theoretical Biology (Gerd Müller, Head), University of Vienna, Althanstrasse 14, 1090 Vienna, Austria. stephan.handschuh@univie.ac.at
authors.forename: Stephan
authors.initials: S
authors.lastname: Handschuh
authors.forename: Thomas
authors.initials: T
authors.lastname: Schwaha
authors.forename: Brian D
authors.initials: BD
authors.lastname: Metscher
mesh_terms: D000489:Alligators and Crocodiles / Q000196:embryology; D000818:Animals; D002013:Bryozoa / Q000166:cytology; D016591:Histocytological Preparation Techniques / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007448:Invertebrates / Q000166:cytology; D008853:Microscopy
references.citation: Dev Biol. 2001 Apr 15;232(2):458-70
references.pmid: 11401405
references.citation: Nephron Exp Nephrol. 2006;103(2):e50-4
references.pmid: 16543764
references.citation: Nat Biotechnol. 2000 Mar;18(3):321-5
references.pmid: 10700150
references.citation: Annu Rev Biomed Eng. 2004;6:209-28
references.pmid: 15255768
references.citation: BMC Biol. 2007 Aug 17;5:34
references.pmid: 17705855
references.citation: Dev Dyn. 2002 Nov;225(3):369-75
references.pmid: 12412023
references.citation: BMC Dev Biol. 2008 Sep 15;8:83
references.pmid: 18793391
references.citation: Anat Rec. 1997 Aug;248(4):583-602
references.pmid: 9268147
references.citation: Microsc Res Tech. 2005 Aug 1;67(5):227-39
references.pmid: 16170824
references.citation: BMC Neurosci. 2004 Aug 06;5:27
references.pmid: 15298700
references.citation: J Microsc. 2007 Jul;227(Pt 1):51-71
references.pmid: 17635659
references.citation: J Neurosci Methods. 2002 Oct 15;120(1):11-6
references.pmid: 12351202
references.citation: Dev Dyn. 2009 Mar;238(3):632-40
references.pmid: 19235724
references.citation: Semin Cell Dev Biol. 1997 Oct;8(5):509-17
references.pmid: 9441956
references.citation: Nat Genet. 2000 Jun;25(2):147-52
references.pmid: 10835627
references.citation: ScientificWorldJournal. 2007 May 11;7:592-604
references.pmid: 17525824
references.citation: J Morphol. 2008 May;269(5):594-603
references.pmid: 18085565
references.citation: Dev Dyn. 2006 Sep;235(9):2386-400
references.pmid: 16871621
references.citation: Anat Rec (Hoboken). 2007 Mar;290(3):277-83
references.pmid: 17525936
references.citation: PLoS Genet. 2006 Apr;2(4):e61
references.pmid: 16683035
references.citation: Microsc Res Tech. 2008 Jan;71(1):51-9
references.pmid: 17868133
references.citation: Nat Genet. 2002 Jan;30(1):59-65
references.pmid: 11743576
references.citation: BMC Physiol. 2009 Jun 22;9:11
references.pmid: 19545439
references.citation: Stain Technol. 1952 Jan;27(1):61-4
references.pmid: 14931079
references.citation: Microsc Res Tech. 2008 Nov;71(11):778-86
references.pmid: 18785246
references.citation: Arthropod Struct Dev. 2008 Mar;37(2):141-54
references.pmid: 18089134
references.citation: Nat Rev Mol Cell Biol. 2003 Sep;Suppl:SS10-6
references.pmid: 14587521