Automated techniques for the study of lung alveolar stereological parameters with the IBAS image analyser on optical microscopy sections

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Abstract
In the case of lung alveoli, the good contrast of the features allows the use of a fully autuomated image analyzing procedure, yielding many of the desirable stereological parameters which can be estimated on optical microscopy slides: alveolar volume and surface densities, a form factor and average mean integral curvature. With the program, specially designed for the IBAS image analyzer (Kontron Bildanalyse, Munich, B.R.D.), the whole automated process takes an average of 28 s/field.The image analyzer-obtained stereological estimates compare well with those given by point and intersection counting methods. Some difficulties inherent in image analyzers arise when measuring alveolar boundary lengths. A scheme is therefore proposed which eliminates the image-edge errors, with no sampling bias, by comparing 3 different frames inside each field. A solution to the problem of estimating automatically the positive and negative tangent counts, necessary for the curvature calculations, is also proposed. This program was applied routinely to the study of premature newborn rabbit lung alveoli.