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Biomedical subjects

B Minnich

Publications and source records attributed to B Minnich.

5 recordsLinked to original sources

Spatial growth and pattern formation in the small intestine microvascular bed from larval to adult Xenopus laevis: a scanning electron microscope study of microvascular corrosion casts.

The microvascular anatomy of the small intestine of metamorphosing tadpoles of the South African Clawed Toad, Xenopus laevis (Daudin) is studied from developmental stages 55 to 65 and in adults by scanning electron microscopy (SEM) of vascular corrosion casts (VCCs) and light microscopy. Up to stage 62, VCCs reveal a dense two-dimensional vascular network ensheating the intestinal tube, whose proximal portion forms a clockwise spiralling outer and its distal portion an anti-clockwise spiralling inner coil. Vessels of the intestinal network impose flat and run circularly to slightly obliquely. Locally, dense capillary plexus with small "holes" indicating ongoing intussusceptive microvascular growth (IMG) and vessel maturation, are present. The typhlosole, an invagination along the proximal portion of the small intestine, reveals a dense capillary bed with locally ongoing IMG. VCCs of stages 62/63 for the first time reveal a three-dimensional vascular bed with longitudinal intestinal folds of varying size and heights greatly enlarging the luminal exchange area of the intestinal tube. From stage 65 onwards, longitudinal intestinal folds undulate and, though smaller in size and less mature as indicated in VCCs by the presence of wider, sinus-like vessels with small "holes" interposed between, closely resemble the intestinal folds present in the small intestine of adult Xenopus. Our data suggest that maturation of the vascular pattern in the small intestine of X. laevis tadpoles takes place successively after stages 62-63, and growth during this period is preferentially by intussusception.

Age Factors↗

Analysis of microvascular trees by means of scanning electron microscopy of vascular casts and 3D-morphometry.

Arterial and capillary trees form by consecutive branching (mostly bifurcations) from a stem vessel, venous trees form by repeated merging of blood vessels. Diameters of stem (parent, mother) vessels and daughter vessels (branches), interbranching distances and branching angles between stem and daughter vessels lastly define the overall three-dimensional structure of the vascular network as well as the basic transport capacity of the system. Here we use scanning electron microscopy and 3D-morphometry to measure these variables from stereo paired images of vascular corrosion casts of the anterior cerebral artery and its main branches and from arteriolar bifurcations of the mesencephalic optic tectum in the actinopterygian fish, Acipenser ruthenus. We then calculate bifurcation indices, area ratios, asymmetry ratios and test for the optimality principles underlying the bifurcations studied. Our results show that arteriolar bifurcations in the optic tectum are in favor of the principles of minimum pumping power and minimum volume rather than the principles of minimum surface and minimum drag. We conclude that scanning electron microscopy of vascular corrosion casts in conjunction with 3D-morphometry is an excellent tool to thoroughly analyze vascular trees in healthy and diseased tissues and organs, as well as on an ontogenetic and phylogenetic scale.

Algorithms↗

Quantitative microvascular corrosion casting by 2D- and 3D-morphometry.

As a system of tubes (blood vessels) the cardiovascular system changes actively and passively diameters to adapt its transport capacities for respiratory gases, nutrients, heat, metabolites and waste products to and off the body's organs, tissues and cells. In most healthy organs blood vessels form a hierarchically arranged three-dimensional network with the geometry defined by vessel diameters, interbranching distances (defining branching frequencies and number of branching sites, i.e. nodes), intervascular distances, and branching angles. In the present study 2D- and 3D-morphometry is applied to quantify these parameters and their changes as they occur in resin casts during metamorphosis of the tadpole lung (2D-morphometry) and filter apparatus vasculature (3D-morphometry). It is shown that 2D-morphometry should be limited to the analysis of high powered images of flat two-dimensional vascular networks (example: tadpole lung alveolar vascular bed) to prevent underestimation of parameters. In contrast, 3D-morphometry can be applied over a wide range of magnifications whereby accuracy of measurements increases with the portion the structure to be measured occupies within the field of view. Together with a careful control of precasting conditions (application of vasoactive drugs, anaesthetics), casting conditions (pressure during rinsing and casting, amount of final shrinkage of casting media), and postcasting conditions (thermal burdening during maceration, sputtering, evaporation, and SEM inspection; thickness of conductive metal layers) 3D-morphometry enables to gain reliable data from resin casts of highly complex real vascular networks in healthy and diseased organs in the developing, juvenile, adult and aged state, as well as in different physiological states.

Aging↗

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.

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 (xL/yL; xR/yR), (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 microm wide square meshes, polystyrene beads with 0.33 microm 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).

Image Processing, Computer-Assisted↗

Lengths measurements in microvascular corrosion castings: two-dimensional versus three-dimensional morphometry.

In the present study we compared measurements of vessel lengths from (a) single-digital scanning electron microscope (SEM) images of microvascular corrosion casts (VCCs) of gill filters of tadpoles of Xenopus laevis Daudin by two-dimensional (2-D) morphometry (Optimas 6.5, Optimas Corp., Bothell, Wash., USA; planar measurements) and (b) digital stereopairs by three-dimensional (3-D) morphometry (3D-Morphometry, Minnich and Muska OEG, Salzburg). Depending on the spatial orientation of the vessels measured, we found a maximum difference of 58.84% (100 [3-D]-41.16 [2-D]) in vessel lengths by 3-D morphometry versus 2-D morphometry, which, in multiple (segmental) lengths measurements or when determining space angles, might be even higher. Based on results we consider 3-D morphometry of VCCs to be the method of choice for lengths measurements.

Animals↗