A Hematoxylin Staining Procedure for Maize Pollen Grain Chromosomes

Studies of postmeiotic chromosome behavior have been impeded by the thick exine and abundant starch grains of maize pollen. Staining pollen grain chromosomes with acetocarmine is tedious and gives inconsistent, often unsatisfactory results. A hematoxylin stain, used in conjunction with the clearing agent chloral hydrate, has been successfully used by the authors to stain chromosomes, nuclei and sperm cells of the maize pollen grain. An ethanol-formaldehyde fixing fluid is used to fix and preserve the pollen samples. The procedure, which is rapid and simple, gives excellent preparations with both fresh and fixed material. Stained preparations do not get darker with time, as is typical of other hematoxylin stained materials.     

A Procedure for Staining Pollen Nuclei When Obscured by Cytoplasmic Inclusions

The number and shape of pollen nuclei in cytologically refractive plants can be determined from anthers fixed in FAA, preserved in 70% ethanol, treated 10-30 min with 50% HCl in 95% ethanol, and stained with iron acetocarmine saturated with chloral hydrate, 1-16 hr. Pollen is then pressed from the anther into a drop of plain acetocarmine, warmed, examined and permanently mounted in Venetian turpentine medium. This procedure clears the cytoplasm of many opaque obstructions and frequently separates the exine from the cytoplast. It may be used with flowering material which has been stored several years at room temperature.     

Staining Pollen Nuclei with Snows Alcoholic Hydrochloric Acid-Carmine

Nuclear staining of pollen grains is improved by using an alcoholic HCl-car-mine stain and Hoyer's mediem, a chloral hydrate-gum arabic mountant. This mountant completes differentiation of the pollen and clears cytoplasmic inclusions which may otherwise obscure the nuclei. Fixing anthers in 3:1 ethanol: acetic acid is recommended. Staining time at room temperature varies between 1 and 5 days, but can be shortened considerably by heat. Pollen is differentiated in 45% acetic acid-addition of Hoyer's medium makes the preparation permanent. The darkening of cytoplasm frequently found in acetocarmine slides does not occur. Pollen from carefullydried plants has given satisfactory staining and makes the method of geater use to taxonomists.     

A Method for Staining Pollen Tubes in Pistil

A quadruple staining procedure has been developed for staining pollen tubes in pistil. The staining mixture is made by adding the following in the order given: lactic acid, 80 ml; 1% aqueous malachite green, 4 ml; 1% aqueous acid fuchsia, 6 ml; 1% aqueous aniline blue, 4 ml; 1 % orange G in 50% alcohol, 2 ml; and chloral hydrate, 5 g. Pistils are fixed for 6 hr in modified Carnoy's fluid (absolute alcohol:chloroform:glacial acetic acid 6:4:1), hydrated in descending alcohols, transferred to stain and held there for 24 hr at 45±2 C They were then transferred to a clearing and softening fluid containing 78 ml lactic acid, 10 g phenol, 10 g chloral hydrate and 2 ml 1% orange G. The pistils were held there for 24 hr at 45±2 C, hydrolyzed in the clearing and softening fluid at 58±1 C for SO min, then stored in lactic acid for later use or immediately mounted in a drop of medium containing equal parts of lactic acid and glycerol for examination. Pollen tubes are stained dark blue to bluish red and stylar tissue light green to light greenish blue. This stain permits pollen tubes to be traced even up to their entry into the micropyle.     

A Rapid and Simple Method for Staining Lipid in Fixed Nematodes

A method is described for staining lipid in fourth-stage dispersal juvenile nematodes fixed with formal-acetic fixative (FA4:1). Bursaphelenchus xylophilus fourth-stage dispersal juveniles were fixed with hot FA4:1 for 24 hours, excess fixative was removed, and a solution of saturated oil red O in 96% ethanol added and allowed to sit for 25 minutes at 60 C. Excess oil red O was removed, nematodes were washed twice with 70% ethanol, and were processed to pure glycerin. Lipid droplets within the nematodes were viewed by light microscopy and appeared as dark red spheres of various sizes. Computerized image analysis was used to quantify lipid droplet area.     

Staining Germinating Pollen and Pollen Tubes

Germinating pollen on stigmas and pollen tubes in styles of Antirrhinum, Brassica, Oenothera, Raphanus, Rosa, solatium and Tagetes spp. were prepared for examination as follows: The styles were fixed in ethyl alcohol-acetic acid 3:1 for 1 hr, and hydrolyzed at 60°C for 5 to 60 min (depending on the species) in 45% acetic acid. The stigma with its attached strand(s) of stigmatoid tissue was then dissected out under a stereoscopic microscope, placed in a few drops of a staining solution made by dissolving 150 mg of safranin O and 20 mg of aniline blue in 25 ml of hot 45% acetic acid. After 5-15 min in this stain, the tissue was placed in a fresh drop of stain on a microscope slide and gently squashed under a cover glass. Because of a gradual precipitation of the aniline blue component, the stain had to be filtered regularly before use. However, a staining solution could be kept at room temperature for several weeks.     

一种简便经济的凝胶电泳同工酶特异染色方法

介绍一种简单、经济的同工酶染色方法：用熔化的0．4％琼脂糖处理滤纸备用,染色前将滤纸浸于同工酶染色液中,染色时将滤纸盖在聚丙烯酸胺胶上,然后将胶放在有盖塑料盒中保温染色,染色时间要比普通方法略长。染色后将胶和滤纸移入固定液中用镊子除去滤纸．     

供光学显微镜观察的花粉样品制备的一种简单方法

传统的制备供光学显微镜观察的花粉样品的方法 (中国科学院植物研究所形态室孢粉组 ,1 960 )不但程序复杂 ,而且不同种类的花粉容易混杂。最近 ,我们在进行山茶属(Ｃａｍｅｌｌｉａ)花粉形态的系统研究中总结出一种制备供光镜观察的花粉材料的简单方法 ,现将其过程介绍如下 :( 1 )从标本或新鲜植株上取下花药 ,用冰醋酸浸软后 ,置洁净的凹玻片 (单凹玻片 )上 ,于解剖镜下将花药打开 ,滴上 95%酒精将花粉洗出。( 2 )滴上预先配制好的分解液 (醋酸酐 9份和浓硫酸 1份 ) ,于室温下或 50℃恒温箱里放置 5ｍｉｎ(具体温度和时间因花粉种类而异 ) …     

A Simple Method for Staining Fungal Substrate Hyphae in Agar Media

Morphogenetic processes often occur in fungal cultures in agar medium. These processes are difficult to study by light microscopy because the hyphae or other structures fail to have sufficient contrast for detailed study and photography. To overcome this difficulty, we developed a method to stain hyphae inside the agar without affecting the medium itself.     

一种简便的考马斯亮蓝G250蛋白质染色方法

介绍一种快速、简便、几乎无背景的考马斯亮蓝G250(CBB G250)染色方法.该方法所用试剂仅为稀盐酸和CBB G250, CBB G250的工作浓度为0.0015%,灵敏度达0.02 μg/带, 染色2 h达70%,4 h以上或染色过夜即可充分染色.与以往的考马斯亮蓝染色方法相比,该方法有经济方便、灵敏度高、几乎无背景等优点,便于推广应用.     

Methenamine-Silver Staining: a Simple and Sensitive Staining Method for Senile Plaques and Neurofibrillary Tangles

An improved methenamine-silver impregnation method is presented which exhibits sensitivity for amyloid substances comparable to that of anti-β protein immunostaining. In optimally treated sections, this technique stained both β-amyloid deposits and neurofibrillary tangles, which are known to have a β-pleated structure. This simple procedure allows a large number of sections to be stained for routine examination.     

水稻原生质体细胞核及原生质体融合体的简易染色观察法

筛选出一种荧光染料罗丹明B(Rhodamine B),利用该荧光染料染色,原生质体细胞核在普通光学显微镜或荧光显微镜下呈红色或发出强烈的桔红色荧光,能清晰地进行分辨。利用罗丹明B或者使用荧光染料FDA,对两种不同来源的原生质体进行染色,在荧光显微镜下,两种不同来源的原生质体分别发出桔红色或绿色荧光,因此可以用于原生质体融合中不同的融合体类型的观察和分析。