A random arrangement of albumin-containing hepatocytes seen with histo-immunologic methods. II. Conditions that produce the artifact.

The cellular immunolocalization of albumin in rat liver has been studied as a function of various physiological and physical conditions. Our observations show that the prime requisite for accurate immunolocalization of albumin and other hepatic-based proteins is the complete removal of blood and especially plasma from sinusoids and the perisinusoidal space of Disse prior to fixation. Fixation of blood-filled liver specimens results in the antifactual entrance of plasma constituents into hepatocytes. When the fixative used in formaldehyde, the artifactual uptake occurs primarily into hepatocytes that have a high glycogen content. Fixation of blood-filled liver with acetic acid-ethanol causes a massive influx of plasma into all hepatocytes. On the contrary, with blood-free liver, varying the type of fixative consistently demonstrates that all hepatocytes normally contain albumin, transferrin, and fibrinogen simultaneously. Increasing the time between cessation of blood flow and outright fixation by either withholding the fixative or by impeding its diffusion through the specimen causes a progressive loss of antigenicity of albumin. The same result ensues when specimens remain in contact with the fixative for an extended time.