Clearing and Staining Plant Material in Toto with Phloroglucinol-Hcl in Methyl Benzoate for Projection Photography and Subsequent Serial Sectioning

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Abstract
Herbaceous plant material is fixed in a mixture of formaldehyde, acetic acid, propionic acid, and ethanol, dehydrated in an ethanol series, cleared in methylbenzoate and stained with phloroglucinol and HCl in methyl benzoate. A picture of the topography of the lignified elements is made by projection photography. The material is then still in good condition for microscopical preparations. Next, the methyl benzoate is washed out with tertiary butanol and the specimens are processed through paraffin, cut, stained with safranin and fast green and mounted. In this way, both an over-all longitudinal photograph and a complete series of cross sections can be obtained from one and the same vascular complex. The method also enables exact orientation of the material for the preparation of longitudinal sections through bundles of vascular tissue whose location cannot be determined externally.