IngentaConnect Three-dimensional morphometry in scanning electron microscopy: a ...

Authors: Minnich¹; Leeb²; Bernroider³; Lametschwandtner⁴

Source: Journal of Microscopy, Volume 195, Number 1, 1 July 1999 , pp. 23-33(11)

Abstract:

A method for accurate dimensional and angular measurements of microstructures analysed in the scanning electron microscope is described. The method considers central and parallel projections and involves (a) digital image acquisition of stereopaired images from the scanning electron microscope's photodisplay, (b) generation of 3D-image representations, (c) setting of measuring points in the digitized stereopaired images, (d) computation of exact space coordinates (x/y/z) from the corresponding point coordinates (x_L/y_L; x_R/y_R), (e) determination of distances and angles between consecutive corresponding points using vector equations, and (f) transfer of computed data into spreadsheets of the data analysis software using dynamic data exchange with simultaneous graphical display of the frequency distribution of variables.

Measurements performed on specimens with known dimensions (grid with 10 μm wide square meshes, polystyrene beads with 0.33 μm diameter) and angles (synthetic crystals of K(Al,Cr)[SO₄], CuSO₄.5H₂O and NaCl) revealed a high accuracy in dimensional as well as angular measurements (total error 1 ± 0.5%).

In Monte Carlo experiments the overall error was found to depend strongly on the size of the measured structure relative to the size of the measurement field (field width).

Keywords: 3D morphometry; digital image analysis; quantification of microstructures; SEM

Document Type: Research article

DOI: 10.1046/j.1365-2818.1999.00478.x

Affiliations: 1: Department of Electron Microscopy, Institute of Zoology, University of Salzburg, Hellbrunnerstr. 34, A-5020 Salzburg, Austria, 2: Institute of Mathematics, University of Salzburg, Hellbrunnerstr. 34, A-5020 Salzburg, Austria, 3: Institute of Information Processing, Department for Applied Computer Science, Vienna University of Economics and Business Administration, Augasse 2-6, A-1090 Vienna, Austria, 4: Department of Experimental Zoology, Institute of Zoology, University of Salzburg, Hellbrunnerstr. 34, A-5020 Salzburg, Austria

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Three-dimensional morphometry in scanning electron microscopy: a technique for accurate dimensional and angular measurements of microstructures using stereopaired digitized images and digital image analysis