A Comparison of Scanning and Two-Wavelength Microspectrophotometry

Abstract

Because of the absence of suitable standards, microspectrophotometry suffers from a lack of objective verification. An approach to this problem is suggested which is based on the comparison of results obtained when different techniques or instruments are applied to the same objects. The application of this approach to scanning versus two-wavelength photometry has been justified by the widely different bases of these two methods. A group of ascites tumor cells stained with gallocyanin-chrome alum was measured successively with both methods and a high degree of proportionality between the two sets of results was found. This suggests that the two methods measure the same quality of the cells within a standard deviation of 3.39 per cent. This degree of correlation is a verification of the accuracy of both of the methods and shows that either one is suitable for resolving differences in stain content between cell nuclei of the order of 10 per cent.