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Biology Faculty Publications Biology

Fall 1999

Three-Dimensional Reconstructions of TadpoleChondrocrania from Histological SectionsGary P. Radice

Mary Kate Boggiano

Mark Desantis

Peter M. Larson

Joseph Oppong

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Recommended CitationRadice, Gary, Mary Kate Boggiano, Mark DeSantis, Peter Larson, Joseph Opping, Matthew Smetanick, Todd Stevens, James Tripp,Rebecca Weber, Michael Kerckhove, and Rafael O. de Sá. "Three-Dimensional Reconstructions of Tadpole Chondrocrania fromHistological Sections." Virginia Journal of Science 50, no. 3 (Fall 1999): 227-35.

AuthorsGary P. Radice, Mary Kate Boggiano, Mark Desantis, Peter M. Larson, Joseph Oppong, Matthew T.Smetanick, Todd M. Stevens, James Tripp, Rebecca A. Weber, Michael Kerckhove, and Rafael O. de Sá

This article is available at UR Scholarship Repository: http://scholarship.richmond.edu/biology-faculty-publications/82

Virginia Journal of Science Volume 50. Number 3 Fall 1999

Three-Dimensional Reconstructions of Tadpole Chondrocrania from Histologic2l Sections

Gary Radice*, Mary Kate Boggiano+, Mark Desantis+, Peter Larson*, Joseph Oppong +.f Matthew Smetanick*

Todd Stevens+, James Tripp , Rebecca Weber*, Michael Kerckhove +,and Rafael de Sa*

*Department of Biology and +Department of Math and Computer Sciences, University of Richmond, Richmond, VA 23173

ABSTRACT Reconstructing three dimensional structures (3DR) from histological sections has always been difficult but is becoming more accessible with the assistance of digital imaging. We sought to assemble a low cost system using readily available hardware and software to generate 30R for a srudy of tadpole chondrocrania. We found that a combination of RGB camera, stereomicro­scope, and Apple Macintosh PowerPC computer.; running NIH Image. Object Image, Rotater. and SURFdri\'er software pro,·ided acceptable reconstruc­tions. These are limited in quality primarily by the distortions arising from histological protocols rather than hardware or software.

INTRODUCTION It has always been difficult to ,;sualize and analyze three dimensional (30 )

histological features, when starting with two dimensional (20) sections. Reconstruct­ing the third dimension from slices presents problems in section regisLration as well as presenting the data in a way that can be readily interpreted. Nevertheless, there are good reasons fo r wanting to \'isualize the third dimension. In our case we wanted to compare the shapes of tadpole chondrocrnnia to find character.; that may be useful in phylogenetic srudies. For this project we sought to develop a system for generating three dimensional reconstructions (30R) using relatively simple. low cost. and widely available hardware and software. Here. '' e summarize our effort and discuss the advantages and pitfalls of digital histological reconstructions.

tv1ETHODS

Specimens Tadpoles from various species \\Cre used. including Hy/a picta. Leptodactylus

gracilis. Phy/10111edusa h_ipochondrialis. Ptychohy/a salvadorensis, Smilisca phaeota. Tripion petasatus. and Xenopus lne1•is. Swimm.ing tadpoles at various stages were k.illed in 3 aminobenzoic acid et11yl ester (MS 222) and subsequently fixed and stored in~% fonnaldehyde, 0.01 M sodium phosphate at pH7.2-7A. Most specimens were fixed after remo,·ing their bod~ Ca\'it~ organs. to speed fixative penetration.

Histology For embedding. specimens were first washed m water, dehydrated through increas­

ing concentrations of ethanol, cleared with Hemo-De® (Fisher Scientific) and embed­ded in ParaPlast Plus® (Oxford Laboratories). For sectioning, specimen blocks were