Abstract
A significant problem in the application of absorption spectrophotometry to tissue sections is that the chromo-phores are often not uniformly distributed; thus, the Beer-Lambert law cannot be used to relate transmittance to the quantity of absorber. There are three methods to compensate for the resulting distributional error: (i) The conventional method, which is suited for large objects, requires measurements be made only over homogeneous regions. (ii) The scanning method (1) integrates the results of many measurements each of which is made over an area sufficiently small so that the absorber distribution can be assumed to be homogeneous. (iii) The two-wavelength method requires measurement of the transmittance at two wavelengths for each field; the mass of absorber in the field is then calculated using the two-wavelength equations (2,3). It has been shown that the scanning and two-wavelength methods are equally accurate (4).
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