title: Comparative study of freeze-substitution techniques for X-ray microanalysis of biological tissue.
abstract: A comparative study of some aspects of freeze-substitution as a preparative method for X-ray microanalysis of diffusible ions in biological specimens was carried out. Four substitution fluids were compared with each other and with cryosections. As criteria, elemental ratios as determined by X-ray microanalysis of thin sections of mouse pancreas were used. Freeze-substitution for 3 weeks in diethyl ether or for 2 days in tetrahydrofurane gave results comparable to those obtained in cryosections. Slow warming to room temperature after freeze-substitution in tetrahydrofurane gave slightly better results than fast warming. Freeze-substitution in acetone resulted in noticeable loss and redistribution of ions and freeze-substitution in methanol gave very poor results.
authors.affiliation: Department of Human Anatomy, University of Uppsala, Sweden.
authors.forename: E
authors.initials: E
authors.lastname: Pålsgård
authors.forename: U
authors.initials: U
authors.lastname: Lindh
authors.forename: G M
authors.initials: GM
authors.lastname: Roomans
mesh_terms: D000818:Animals; D004577:Electron Probe Microanalysis; D017110:Freeze Substitution / Q000379:methods*; D008297:Male; D051379:Mice; D008820:Mice, Obese; D010179:Pancreas / Q000737:chemistry* / Q000648:ultrastructure