The use of freezesubstitution and lr gold in the study of rye grass (Lolium perenne) pollen

Liza B. Martinez, Sue M Wick

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6 Scopus citations


Pollen grains of Lolium perenne (rye grass) were prepared for transmission electron microscopy by rapid freezing in liquid propane, substitution in acetone, methanol or diethyl ether, and embedment in the acrylic resin London Resin gold. These were compared to pollen chemically fixed (CF) in aldehyde/osmium tetroxide and embedded in the epoxy resin Quetol 651. Ultrastructural preservation was superior in freezesubstituted (FS) pollen, particularly with the use of acetone or methanol. Optimally preserved FS pollen displayed a homogeneous aspect of the cytoplasm and nucleoplasm, and smooth, uninterrupted contour or organelles. A striking difference was also seen in the preservation of inclusions in the intine. Varied forms and sizes of intine inclusions were evident in FS pollen but these were not discernible in the CF image. The FS scheme studied here presents enormous potential for both ultrastructural and immunolabelling studies in rye grass pollen. Problems discussed include artifacts associated with each of the substitution solvents used, and a gradient of freezing damage observed within the pollen grain. Copyright1991 WileyLiss, Inc.

Original languageEnglish (US)
Pages (from-to)305-314
Number of pages10
JournalJournal of Electron Microscopy Technique
Issue number3
StatePublished - Jul 1991


  • Acetone
  • Chemical fixation
  • Diethyl ether
  • Freezing artifact
  • Methanol

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