Abstract
A computational procedure and a software
program were developed for three-dimensional (3D)
modeling of the rice embryo and of the gene
expression of the homeobox gene OSH1. To date,
such samples have generally been difficult to analyze
three-dimensionally by confocal laser microscopy.
Therefore, we performed 3D reconstruction using
serial sections. The procedure can be summarized
as follows. Serial sections were subjected to in situ
hybridization with subsequent digitization of
resultant micrographs. Then, two-dimensional (2D)
images of sections were realigned by automatic or
interactive computation to restore their original 30
positions, which had been destroyed by sectioning.
After realignment, data relating to all sections of
the embryo and gene expression were extracted
according to color differences defined by an HSL
(hue, saturation, and lightness) coordinate system
with subsequent noise cancellation. The sectional
contours of the various regions were connected to
generate 30 surfaces by triangulation. Each 3D
triangulated frame was subjected to a rendering
program, and finally a 3D model of the rice embryo
and of expression of the gene examined by in situ
hybridization was generated. The resultant 30
model clearly demonstrated down-regulation of
expression during the development of the coleoptile
and provascular tissue, confirming the usefulness of
our 30 modeling technique.
