An Analysis of Methods for Permanently Mounting Ovules Cleared in Four-and-a-Half Type Clearing Fluids
- 1 January 1982
- journal article
- research article
- Published by Taylor & Francis in Stain Technology
- Vol. 57 (3), 161-169
- https://doi.org/10.3109/10520298209066609
Abstract
Ovules cleared in benzyl benzoate-4 1/2 clearing fluid can be permanently mounted in Piccolyte or Permount by replacing the clearing fluid with absolute ethanol, upgrading the ovules in mixtures of ethanol and xylene (3:1, 2:2, 1:3, and xylene), and mounting them in either mountant under the supported coverglass of a Raj slide. Optical sagittal sections through the ovules resemble microtome sections in that the protoplasts are slightly shrunken away from the cell walls. The artifact is common in permanently mounted sections; fixation and paraffin infiltration are usually cited as the causes—its appearance in the whole-mounted ovules is caused by xylene. Although miscible with the clearing fluid, Euparal is the least satisfactory of the standard mountants for permanent preparations of cleared ovules and is best used with an equal quantity of clearing fluid for semipermanent preparations. A large quantity of Euparal in the mountant produces pronounced shrinkage. A method for permanently mounting cleared ovules with the clearing image unaltered employs a mountant which contains the ingredients of Spurr low viscosity embedding medium. Vinylcyclohexene dioxide (10 drops) is combined with diglycidyl ether of polypropylglycol (6 drops) and nonenyl succinic anhydride (26 drops). Ovules treated for 24 hr in benzyl benzoate-4 1/2 clearing fluid are passed through a graded series of clearing fluid-epoxy medium mixtures (3:1, 2:2, 1:3, and pure epoxy medium) at intervals of 15 minutes. One drop of dimethylaminoethanol, the cure accelerator, is then added to the epoxy medium and the ovules are mounted and covered immediately on a Raj slide. The preparation is cured in an oven at 60 C for 24 hr and observed with phase contrast or Nomarski interference optics.This publication has 23 references indexed in Scilit:
- Raphide Files in Vegetative Organs of ZebrinaBotanical Gazette, 1979
- A Nomarski Interference Study of Megasporogenesis and Megagametogenesis in Smelowskia calycina (Cruciferae)American Journal of Botany, 1978
- Anatomy of Maize Tissue CulturesPhysiologia Plantarum, 1978
- A Method of Permanently Mounting Biological Tissue Cleared in Herr's Four-And-A-Half Clearing FluidStain Technology, 1976
- Infraspecific polyploidy and gynodioecism in Ptilotus obovatus (Amaranthaceae)Australian Journal of Botany, 1976
- The Gametophytes of Ophioglossum palmatum L.American Journal of Botany, 1975
- Plantago WiltPhytopathology®, 1975
- An Overview of Botanical Clearing TechniqueStain Technology, 1975
- The Use of Herr Four-and-a-Half Clearing Fluid for the Rapid Microscopic Examination of Thick Sections of Normal and Neoplastic TissuesStain Technology, 1975
- Physiological activity of corticosteroids in etiolated Mung bean plantsZeitschrift für Pflanzenphysiologie, 1974