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. 1972 Feb;49(2):166–171. doi: 10.1104/pp.49.2.166

The Retention of Water-soluble Compounds during Freeze-Substitution and Microautoradiography 1,2

Donald B Fisher a, Thomas L Housley a
PMCID: PMC365921  PMID: 16657917

Abstract

Freeze-substitution and Epon embedment were quantitatively evaluated for their effectiveness in retaining water-soluble metabolites in plant tissues. Roughly 99% of the 80% (v/v) ethanol-extractable radioactivity in photosynthetically labeled soybean leaf discs and in petiole fragments containing translocated 14C was retained during freeze-substitution in acetone or propylene oxide and embedment in Epon. Substantially more activity was lost from 14C-sucrose-infiltrated pith blocks, but most or all of this loss came from the block surface. The procedure was effective for a sucrose concentration as low as 0.004%. Sections floated on water retained most of their 14C-sucrose, and high resolution autoradiographs could easily be prepared without resorting to dry procedures. Embedded 14C-sucrose was apparently chemically unreactive, since there was no loss of radioactivity when sections were stained with the periodic acid-Schiff reagent, nor did the embedded sucrose show staining.

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Selected References

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  1. BRANTON D., JACOBSON L. Dry, high resolution autoradiography. Stain Technol. 1962 Jul;37:239–242. doi: 10.3109/10520296209117744. [DOI] [PubMed] [Google Scholar]
  2. CARO L. G., VAN TUBERGEN R. P., KOLB J. A. High-resolution autoradiography. I. Methods. J Cell Biol. 1962 Nov;15:173–188. doi: 10.1083/jcb.15.2.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fisher D. B. Kinetics of C-14 translocation in soybean: I. Kinetics in the stem. Plant Physiol. 1970 Feb;45(2):107–113. doi: 10.1104/pp.45.2.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. LEPAGE M. THE SEPARATION AND IDENTIFICATION OF PLANT PHOSPHOLIPIDS AND GLYCOLIPIDS BY TWO-DIMENSIONAL THIN-LAYER CHROMATOGRAPHY. J Chromatogr. 1964 Jan;13:99–103. doi: 10.1016/s0021-9673(01)95078-2. [DOI] [PubMed] [Google Scholar]
  5. Stirling C. E., Kinter W. B. High-resolution radioautography of galactose-3H accumulation in rings of hamster intestine. J Cell Biol. 1967 Dec;35(3):585–604. doi: 10.1083/jcb.35.3.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Trip P. Sugar transport in conducting elements of sugar beet leaves. Plant Physiol. 1969 May;44(5):717–725. doi: 10.1104/pp.44.5.717. [DOI] [PMC free article] [PubMed] [Google Scholar]

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