Surface imaging microscopy using an ultramiller for large volume 3D reconstruction of wax- and resin-embedded tissues.
Three-dimensional reconstruction of large tissue volumes using histological thin sections poses difficulties because of registration of sections, section distortion, and the possibility of incomplete data set collection due to section loss. We have constructed an integrated surface imaging system that successfully addresses these problems. Embedded tissue is mounted on a high precision XYZ stage and the upper surface is iteratively: (i) stained to provide an effective optical section, (ii) imaged using a digital camera, and (iii) removed with an ultramiller. This approach provides for the reconstruction of high-quality 3D images by inherently preserving image registration, eliminates section distortion, thus removing the need for complex realignment and correction, and also ensures full capture of all image planes. The system has the capacity to acquire images of tissue structure with voxel sizes from 0.5 to 50 mum over dimensions ranging from micrometers to tens of millimeters. The ultramiller enables large samples to be imaged by reliably removing tissue over their full extent. The ability to visualize key features of 3D tissue structure across such a range of scale and resolution will facilitate the development of a greater understanding of the relationship between structure and function. This understanding is essential for better analyses of the structural changes associated with different disease states, and the development of structure-based computer models of biological function.
Gerneke
Dane A
DA
Bioengineering Institute, The University of Auckland, Auckland, New Zealand. d.gerneke@auckland.ac.nz
Sands
Gregory B
GB
Ganesalingam
Ramanujan
R
Joshi
Purva
P
Caldwell
Bryan J
BJ
Smaill
Bruce H
BH
Legrice
Ian J
IJ
D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation* / Q000379:methods*; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D008867:Microtomy / Q000379:methods; D009206:Myocardium / Q000166:cytology*; D016610:Tissue Embedding