Improved Procedures in Fungal Cytology Utilizing Giemsa

Abstract

To study nuclear events in fructifications of the Basidiomycetes, material was fixed 24 hr in a saturated aqueous solution of HgCl₂ containing 1% glacial acetic acid, and embedded in Aquax (G. T. Gurr Ltd.). Following a 4 hr hydrolysis at 20 C in 60% H₃PO₄, sections were stained for 30 min in a mixture of 4 ml Giemsa R66 (G T. Gurr Ltd.) and 100 ml phosphate buffer at pH 6.5. Differentiation was carried out in sodium cacodylate-HCl buffer at pH 5.8 when required. Preparations were dehydrated in an acetone-xylene series prior to mounting in Euparal. The use of paraffin wax as the embedding medium and HCl as the hydrolysing agent yielded preparations of an inferior quality.