Enzyme cytochemistry of unfixed leukocytes and bone marrow cells using polyvinyl alcohol for the diagnosis of leukemia.
Cytochemical methods for the demonstration of enzyme activities in blood and bone marrow cells were systematically improved by the addition of an inert polymer, polyvinyl alcohol (PVA), to the incubation medium and by using optimized reaction media. The methods investigated were tetrazolium salt methods for lactate, glucose-6-phosphate, succinate and glutamate dehydrogenase, the indoxyl-tetrazolium salt method for alkaline phosphatase, the diaminobenzidine method for peroxidase, and diazonium salt methods for chloroacetate esterase, beta-glucosaminidase, beta-glucuronidase, acid phosphatase, and dipeptidylpeptidase II and IV. PVA in the media preserved the morphology of cells very well and prevented leakage of large molecules such as enzymes from the cells. Therefore, fixation or long periods of air-drying prior to incubation leading to substantial loss of enzyme activity could be avoided. A brief period of drying (2 min at 37 degrees C) of the cell preparations just before the incubation was sufficient for making the cells permeable. Localization of enzyme activities was very precise and precipitation of the final reaction product was confined to sites which are known to contain the enzyme under study (granules, mitochondria, lysosomes). These advantages advocate the use of PVA in haematological enzyme cytochemistry and especially for diagnosis of leukemia.
Van Noorden
C J
CJ
Laboratory of Cell Biology and Histology, University of Amsterdam, The Netherlands.
Vogels
I M
IM
Van Wering
E R
ER
D001853:Bone Marrow / Q000201:enzymology*; D001854:Bone Marrow Cells; D004796:Clinical Enzyme Tests / Q000379:methods*; D005069:Evaluation Studies as Topic; D005404:Fixatives; D006651:Histocytochemistry; D006801:Humans; D007938:Leukemia / Q000175:diagnosis*; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis; D007962:Leukocytes / Q000166:cytology / Q000201:enzymology*; D011142:Polyvinyl Alcohol; D014021:Tissue Preservation / Q000379:methods
D005404:Fixatives; D011142:Polyvinyl Alcohol