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Effect of Fixation on Demonstration of Phosphatases of Eimeria tenella Grown in Chick Kidney Cell Cultures
Author(s) -
VETTERLING JOHN M.,
WALDROP HARRY R.
Publication year - 1976
Publication title -
the journal of protozoology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 0022-3921
DOI - 10.1111/j.1550-7408.1976.tb03794.x
Subject(s) - eimeria , fixation (population genetics) , phosphatase , biology , kidney , microbiology and biotechnology , andrology , biochemistry , medicine , genetics , phosphorylation , gene
SYNOPSIS. The effects of fixation with various concentrations of glutaraldehyde or formaldehyde, acetone or ethanol, and freeze‐drying on 5 phosphatases of Eimeria tenella and chick kidney cell cultures were demonstrated in situ. Gultaraldehyde inactivated the phosphatases more than did the formaldehyde, but the effect of the combination of the 2 (Karnovsky's fixative) was greater than that of either glutaraldehyde or formaldehyde alone. The higher the concentration of aldehyde and the longer the duration of exposure, the greater the inactivation. The order of sensitivity to aldehyde fixation of the enzymes tested was glucose‐6‐phosphatase > thiamine pyrophosphatase > 5′‐nucleotidase > adenosine triphosphatase > acid phosphatase. Cytologic detail was preserved more efficiently with glutaraldehyde than with formaldehyde. Optimal preservation of enzyme activity for cytochemistry was with 2% glutaraldehyde for 30 min or 2% formaldehyde for 1 hr for G‐6‐Pase. TPPase, and 5′‐nucleotidase, and with 2% glutaraldehyde or 2% formaldehyde for 2 hr with ATPase and AcPase. Quenching with subsequent fixation in cold acetone or ethanol resulted in complete inactivation of G‐6‐Pase, TPPase, and 5′‐nucleotidase; although cells fixed in this manner yielded large amounts of reaction product for ATPase and AcPase, the distribution was diffuse, and some of it appeared to be artifactual. Quenching with subsequent freeze‐drying was unsatisfactory because nearly all of the cell layers rolled off the cover glasses.