ENZYMORPHOLOGY: A STUDY OF FREE CELLS

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Abstract
A technique has been developed for the enzymorphologic study of five enzymes in free cells. The material investigated was an ascitic form of a mouse monocytoid leukemia (C-1498), Ehrlich ascites carcinoma and human blood leukocytes. Preservation of the integrity of cell morphology and of enzyme activity was accomplished by the use of a formaldehyde-chloralhydrate fixative followed by embedding in Knox gelatin. On refrigeration, solid gelatin blocks embedding suspended cells result. These may be mounted on a freezing microtome and 5 µ sections readily cut and handled. The location of β-glucuronidase, acid phosphatase, alkaline phosphatase, diphosphopyridine nucleotide-diaphorase and α-naphthyl esterase in these various cells has been described in detail. For β-glucuronidase, diphosphopyridine nucleotide-diaphorase and acid phosphatase, consistent findings of cell enzymorphology are reported for the first time. The technique lends itself to application for all types of cell populations. It also has the great advantage that the same population of cells in gelatin can be subjected to repeated scrutiny over a period of at least three weeks. The conditions of fixation economize on cell material in that a different fixative for each enzyme has become unnecessary, as has also the restriction of working with fresh frozen material prior to performing the reaction for diphosphopyridinenucleotide-diaphorase.