• 作者： 鄭炳今; D. B. Walden; R. I. Greyson
• 作者服務機構： 加拿大西安大略大學植物學系
• 中文摘要： 本文介紹一種高穿透性醛類固定液，對電子顯微鏡標本之固定效果極佳；同時，由於這種固定液之高穿透性，可用以固定質地緊密、體積大或具有表面角質層之植物組織。 在固定液中加以4％鞣酸 (C H 0 ) 作為掃描式電子顯微鏡標本之固定液，並經四氧化鋨(O O ) 再固定之後，可增加標本自身的導電度；如再以鞣酸－三聚甲醛 ((HCHO)　)混合液與四氧化鋨溶液交互處理組織，可大幅地提高標本的導電度，降低標本在掃描電子束轟擊下局部帶電現象；甚至表面未鍍膜的標本亦可直接觀察。 許多組織結構對滲透壓差之改變非常敏感，為減少由脫水及塑膠滲透作業所造成之變形；本文報導一項連續梯度脫水裝置，用以處理大量組織之脫水極為方便；由於裝置極簡易，任何實驗室均可自行裝設。筆者應用此法解決了花葯內小胞子在脫水作業中凹扁變形之困擾。
• 英文摘要： Densely packed or highly cuticularized planttissue does not allow rapid penetration of conven-tional glutaraldehyde fixative, hence poor fixa-tion usually results. Furthermore, air bubbles ortrapped air in plant tissues create an air-tissueinterface which prevents rapid fixative penetra-tion. Fixation, however, can be improved eitherby decreasing the size of the tissue or by usinga highly penetrative fixative. The fixative re-ported in this article was originally developedfor the fixation of whole corn anthers. Its usepermits retention of microspores and preventsdistortion of the tapetal cells which occurs withthe use of conventional fixation methods. Thefixative consists of 0.1% Tween 20 (a wettingagent) and 0-4% sucrose (osmoticum) in 1mlof 40% paraformaldehyde, 97.5ml of 3% glu-taraldhyde (in 0.05 M phosphate buffer, pH7.2),and 1.5ml of acrolein. Tissues were post fixedin O O , sectioned and stained with uranylacetate and lead citrate. The results of thisprocedure are comparable to those obtainedwhen conventional methods are used. SEM specimen were fixed by the samemethod but with 4% tannic acid added. Theywere washed in buffer, post fixed in O O ,treated in 4% tannic acid-paraformaldehydemixture and retreated in O O. The above methodincreases the conductivity of the specimens andprevents local charging phenomena. Surfacecoating is unnecessary if the tannic acid-para-formaldehyde and O O treatment are repeated. Subsequent dehydration and plastic infiltra-tion by conventional methods result in the col-lapse of young microspores. This problem canbe solved by the use of continous-gradient-dehydration and continous-gradient-plastic-infil-tration. In this report, a simple appatus isdescribed for this latter technique.
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