Summary
As a prerequisite for the histochemical study of sequence iron-“hematoxylin” stains the iron alum-acidified hematein procedure was developed which does not require differentiation.
Histochemical blocking and extraction procedures demonstrated that carboxyl and hydroxyl groups are essential for the binding of cationic iron.
The iron alum-Prussian blue reaction colored collagen, reticulum fibers and basement membranes more intensely than muscle fibers. Treatment of tissue sections mordanted in iron alum with the acidified hematein solvent resulted in practically complete removal of iron from all tissue structures. It must therefore be concluded that the selective staining of muscle fibers, terminal bars and related structures with sequence iron-hematein stains is not due to high affinity of iron for these tissue components.
Observations by R. and M. Heidenhain on sequence hematoxylin-potassium dichromate and hematoxylin-alum stains and data from modern textile chemistry indicate that the staining patterns obtained with metal-hematein sequence stains are determined by the affinity of the hematein moiety for certain tissue structures.
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Puchtler, H., Sweat, F. On the mechanism of sequence iron-hematein stains. Histochemie 4, 197–208 (1964). https://doi.org/10.1007/BF00290864
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DOI: https://doi.org/10.1007/BF00290864