A microassay for detection of DNA and RNA in small numbers of plant cells

Summary A microtechnique for the detection of DNA or RNA in small numbers of plant cells (1–50) has been developed using cauliflower mosaic virus (CaMV) infection of turnip as a model system. Both DNA and RNA extracted from 10 mesophyll protoplasts from CaMV-infected plants can be detected by hybridization using a radioactive probe made from cloned CaMV DNA (pCaMV10). No hybridization above background was detected in extracts of protoplasts from uninfected plants. At least 0.15 pg (11 000 molecules) of purified pCaMV10 DNA can be detected. This method is superior to existing macro techniques for nucleic acid detection as smaller amounts of tissue are required and the detection is approximately 100-fold more sensitive. re]19850326 rv]19850530 ac]19850611     

Azophloxine in Two-Dye and Four-Dye Staining Combination

Tissues are fixed in either 10% formalin or Lavdow-sky's mixture. After the tissues are sectioned and mounted, they are stained in hematoxylin, then counterstained for 2 minutes in 0.1% aqueous azophloxine to which 4 drops of acetic acid have been added to each 100 ml. of stain. Sections are then rinsed in 0.2% acetic acid and dehydrated. Azophloxine GA can be used also in a tetrachrome method. Sections are stained in Harris' hematoxylin, washed, and placed in 0.2% acidified aqueous azophloxine for 2 minutes. They are then rinsed in 0.2% acetic acid, stained 1 minute in an aqueous mixture of 4% phosphotungstic acid and 2% orange G solution and rinsed again in 0.2% acetic acid. Finally, they are stained in 0.2% light green for 2 minutes, and differentiated in 0.2% acetic acid for 5 minutes. The advantage in using azophloxine is that it is clear and delicate and when used in a constant concentration, does not overstain if the recommended procedure is followed.     

整体透明技术在植物生物学中的应用实例及其剖析

通过采用不同的透明剂和透明方法,对番茄（Lycopercicon esculenturm）侧根原基、加拿大一枝黄花（Solidago canadensis）与一枝黄花（S．decurens）的亲和性识别反应和苏门白酒草（Conyza sumatrensis）的胚珠发育过程进行了观察,提供了整体透明技术在植物生物学中的应用实例,简要回顾了该技术在植物生殖生物学中的应用和发展状况,分析了该技术在植物生物学应用中的优势和不足,探讨了该技术应用中一些具体的技术环节,如透明剂的选择和使用以及与特殊用途显微镜的配合使用等方面的问题,并对该技术的应用前景进行了展望。     

Methods for taxonomic and distributional studies of terrestrial flatworms (Tricladida: Terricola)

Traditionally used methods for collecting and preserving terrestrial flatworms have proved deficient in various respects. Comparison of methods for preparing these animals for taxonomic study has shown that preservation of morphological characters can best be achieved using formaldehyde-calcium-cobalt fixative and subsequent storage in an aqueous solution of propylene phenoxetol plus propylene glycol and that the best-lasting histological preparations make use of light-fast, oxidation-resistant dyes and synthetic mounting media with an anti-oxidant.     

Chemical Dehydration of Specimens with 2,2-Dimethoxypropane (DMP) for Paraffin Processing of Animal Tissues: Practical and Economic Advantages over Dehydration in Ethanol

Chemical dehydration can be accomplished using 2,2-dimethoxy-propane (DMP). In the presence of an acid catalyst, this liquid reacts with water generating methanol and acetone as products. Although DMP is more expensive per milliliter than ethanol and other solvents used for dehydration, it is an economical alternative because a much smaller volume is needed. Slow penetration of DMP was previously thought to restrict its use to tiny specimens, but we now show that pieces of tissue as thick as 2 cm are dehydrated by overnight immersion in acidified DMP. We also show that dehydration in acidified DMP does not impair the staining of UNA or other basophilic components of animal tissues. The temperature and concentrations of methanol and H⁺ in the chemical dehydrating agent are too low to produce histochemicaUy detectable methylation or nucleic acid extraction.     

Reaction kinetics in biofilms

A novel in situ microtechnique allows evaluating parameters of diffusion-controlled reactions in biofilms. A microprobe, 15 mum in diameter, was used to simultaneously measure the dissolved oxygen concentration and the optical density at different depths in a submerged biofilm. Based on the results, the biofilm diffusion coefficient for dissolved oxygen, D(f) the dissolved oxygen flux through the biofilm surface, J(02), and the half velocity coefficient, K(s), have been calculated.     

A Simple Method for Fixing, Dehydrating and Embedding Pollen Tubes Cultivated in Vitro for Optical and Transmission Electron Microscopy

A simple method to cultivate pollen tubes in a gelatin medium is presented. After the growth of the pollen tubes in the culture medium, they are fixed, dehydrated, and embedded in resin for ultramicrotomy. The method is easy and does not require the purchase of special materials beyond those needed for the usual techniques for studying biological specimens under transmission electron microscopy.