Novel fixation of plant tissue, staining through paraffin with alcian blue and hematoxylin, and improved slide preparation.
Onion (Allium cepa) root tips were fixed in a proprietary solution without aldehyde, toxic metals or acetic acid. Fixed specimens were embedded in paraffin, sectioned on a rotary microtome and mounted on detergent-washed slides without adhesive. Slides with ribbon segments affixed were immersed in 0.2% aqueous alcian blue 8GX in screw-capped Coplin jars in a water bath at 50 C for 1 hr. Excess alcian blue was rinsed off under cold running tap water and the slides were immersed in quick-mixed hematoxylin at room temperature for 15 min. Stained slides were deparaffinized, rinsed with isopropanol, air dried, and coverslips were affixed with resin. Thus, the traditional paraffin microtechnique has been modified at all steps from fixation to finishing slides with coverslips.
Graham
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Department of Ornamental Horticulture, University of Tennessee, Knoxville 37901-1071, USA.
Joshi
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D000423:Alcian Blue; D006416:Hematoxylin; D008867:Microtomy / Q000379:methods; D016612:Paraffin Embedding / Q000379:methods*; D018517:Plant Roots / Q000166:cytology; D010944:Plants / Q000648:ultrastructure*; D013194:Staining and Labeling / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods*
D000423:Alcian Blue; D006416:Hematoxylin