共查询到20条相似文献,搜索用时 31 毫秒
1.
Randall J. Aweele 《Biotechnic & histochemistry》1976,51(1):17-24
A procedure is described in which thick sections (2-10μ or more) of plastic-embedded plant tissues are mounted in serial order on slides for use in routine light microscopy. Sections are cut with a steel knife on a rotary microtome while the block and blade are bathed with 40% alcohol. The cut sections are placed, in order, in 50% alcohol in the small wells of modified plastic trays where they become flat, pliable and suitable for subsequent handling. Sections remain separate and in correct order in the trays while they are stained, washed, and prepared for final mounting on slides. Mounting involves a simple and rapid procedure of transferring the sections to a slide and heating first on a 70-75 C hot plate (to slowly evaporate the water around the section and to partially affix the section) and then on a 100 C hot plate. This second heating ensures adhesion when xylene-base mounting media, which tend to loosen weakly adhered plastic from the slides, are used. The technique of staining the sections loose provides the following advantages: (1) the problems of section loss and entrapment of stain between section and slide during staining are eliminated, (2) relatively high staining temperature, akalinity, and alcohol concentration of the stain solvent (all of which promote loosening of pm-affixed sections from slides during staining) is allowed, and (3) staining is more even and selective. The procedure has been found to be reliable and fast enough to be of value in a significant variety of routine light microscope studies. 相似文献
2.
R J Ameele 《Stain technology》1976,51(1):17-24
A procedure is described in which thick sections (2-10 mu or more) of plastic-embedded plant tissues are mounted in serial order on slides for use in routine light microscopy. Sections are cut with a steel knife on a rotary microtome while the block and blade are bathed with 40% alcohol. The cut sections are placed, in order, in 50% alcohol in the small wells of modified plastic trays where they become flat, pliable and suitable for subsequent handling. Sections remain separate and in correct order in the trays while they are stained, washed, and prepared for final mounting on slides. Mounting involves a simple and rapid procedure of transferring the sections to a slide and heating first on a 70-75 C hot plate (to slowly evaporate the water around the section and to partially affix the section) and then on a C hot plate. This second heating ensures adhesion when xylene-base mounting media, which tend to loosen weakly adhered plastic from the slides, are used. The technique of staining the sections loose provides the following advantages: (1) the problems of section loss and entrapment of stain between section and slide during staining are eliminated, (2) relatively high staining temperature, alkalinity, and alcohol concentration of the stain solvent (all of which promote loosening of pre-affixed sections from slides during staining) is allowed, and (3) staining is more even and selective. The procedure has been found to be reliable and fast enough to be of value in a significant variety of routine light microscope studies. 相似文献
3.
M. P. Alexander 《Biotechnic & histochemistry》1980,55(1):13-18
A versatile stain has been developed for demonstrating pollen, fungal hyphae and spores, bacteria and yeasts. The mixture is made by compounding in the following order: ethanol, 20 ml; 1% malachite green in 95% ethanol, 2 ml; distilled water, 50 ml; glycerol, 40 ml; acid fuchsin 1% in distilled water, 10 ml; phenol, 5 g and lactic acid, 1-6 ml. A solution has also been formulated to destain overstained pollen mounts. Ideally, aborted pollen grains are stained green and nonaborted ones crimson red. Fungal hyphae and spores take a bluish purple color and host tissues green. Fungi, bacteria and yeasts are stained purple to red. The concentration of lactic acid in the stain mixture plays an important role in the differential staining of pollen. For staining fungi, bacteria and yeasts, the stain has to be acidic, but its concentration is not critical except for bacteria. In the case of pollen, staining can be done in a drop of stain on a slide or in a few drops of stain in a vial. Pollen stained in the vial can be used immediately or stored for later use. Staining is hastened by lightly flaming the slides or by storing at 55±2 C for 24 hr. Bacteria and yeasts are fixed on the slide in the usual manner and then stained. The stock solution is durable, the staining mixture is very stable and the color of the mounted specimens does not fade on prolonged storage. Slides are semipermanent and it is not necessary to ring the coverslip provided 1-2 drops of stain are added if air bubbles appear below the coverslip. The use of differentially stained pollen mounts in image analyzers for automatic counting and recording of aborted and nonaborted pollen is also discussed. 相似文献
4.
Plastic sections 0.5 to 2 μm thick are routinely used for light microscopy. Although plastic sections have several advantages over paraffin or celloidin sections, a problem that is often encountered with plastic sections is wrinkling (Fig. 1). Wrinkling occurs during staining when sections dried on glass slides are covered with stain and heated to hasten the penetration of the stain. Mounted sections heated on glass slides, but not stained, ordinarily lack wrinkles, even when examined with phase contrast optics. Similarly, mounted sections covered with stain, but not heated, lack wrinkles; unfortunately, such sections fail to stain adequately. Unmounted sections floated on heated drops of stain also lack wrinkles (Millonig 1980). Thus, it is clear that wrinkling occurs only when mounted sections are covered with stain and heated. 相似文献
5.
Methods are proposed for staining plant chromosomes with the dye brilliant cresyl blue, and for making these stained preparations permanent by using polyvinyl alcohol mounting medium.
The stain, which is composed of 2% brilliant cresyl blue in 45% aqueous acetic or propionic acid, is used with fixed material in making smear preparations. The technics for staining are similar to those employed in the aceto-carmine method.
The mounting medium is made by mixing 56% polyvinyl alcohol, which is diluted in water to the consistency of thick molasses, with 22% lactic acid and 22% phenol by volume. The permanent slides are made by floating off the cover slip of the temporary slide in 70% alcohol, then applying the mounting medium and replacing the cover slip.
The chief advantages of the methods described are:
1)The preparation of the stain is rapid and simple. The batch of stain will be good with the first try.
2)The staining procedure in some instances is shorter than when using aceto-carmine.
3)The stain shows a high degree of specificity for nuclear structures and gives better results than aceto-carmine when used on certain plant tissues.
4)A minimum number of cells is lost in making the slides permanent when using polyvinyl alcohol mounting medium as the slide and cover slip are run through only one solution prior to mounting.
5)The mounting medium dries rapidly and this shortens the time required before critical examination of the permanent mounts can be made. 相似文献
The stain, which is composed of 2% brilliant cresyl blue in 45% aqueous acetic or propionic acid, is used with fixed material in making smear preparations. The technics for staining are similar to those employed in the aceto-carmine method.
The mounting medium is made by mixing 56% polyvinyl alcohol, which is diluted in water to the consistency of thick molasses, with 22% lactic acid and 22% phenol by volume. The permanent slides are made by floating off the cover slip of the temporary slide in 70% alcohol, then applying the mounting medium and replacing the cover slip.
The chief advantages of the methods described are:
1)The preparation of the stain is rapid and simple. The batch of stain will be good with the first try.
2)The staining procedure in some instances is shorter than when using aceto-carmine.
3)The stain shows a high degree of specificity for nuclear structures and gives better results than aceto-carmine when used on certain plant tissues.
4)A minimum number of cells is lost in making the slides permanent when using polyvinyl alcohol mounting medium as the slide and cover slip are run through only one solution prior to mounting.
5)The mounting medium dries rapidly and this shortens the time required before critical examination of the permanent mounts can be made. 相似文献
6.
L. M. A. Lataster A. van Geest H. van den Berge J. Drukker 《Biotechnic & histochemistry》1991,66(2):93-98
Otic bullas of the rat, obtained by excision and formalin fixed, are successfully embedded in methylmethacrylate by dehydration and subsequent infiltration with plastic under vacuum. Sections 10 μm thick are obtained by cutting the trimmed and sandpapered acrylic blocks on an LKB multirange microtome. The sections are collected on adhesive tape and stained with a Trichrome stain (modified Weigert-van Gieson). Finally, the sections attached to the tape are mounted on microscope slides with glycerin-gelatin and sealed in the same medium. Serial sections are used for three-dimensional graphic reconstruction. 相似文献
7.
A technique was developed for restoring broken cytology slides so that they are close to their original condition and for making multiple slides from a single smear preparation. The method is applicable to both cytologic preparations and histologic sections. In this study the fragmented smear preparation was treated with Pro-Texx, which penetrated, impregnated and solidified the full thickness of the pieces of the smear, enabling them to be lifted from the pieces of the broken slide. The removed pieces of the smear preparation were reassembled onto a new slide, which was then restained and coverslipped. In preparing multiple teaching slides, the treated smear preparation was divided as planned, with each portion mounted onto a separate slide, which was then restained and coverslipped. Ten other fine needle aspiration cases with broken slides have been restored, and more teaching slides were prepared from a single smear preparation using the same technique. All were equally successful. This technique provides an excellent method of smear transfer in cases of broken slides and creation of multiple slides from a single smear preparation for cytology teaching. This is particularly useful for unusual cases. 相似文献
8.
A machine is described which can perform the Gram stain. Comparison of slides stained by machine versus hand revealed no difference in reproducibility or accuracy. In addition to providing clean, dry, uniformly stained slides, the machine saves 24 sec per slide when compared with a hand staining technique. 相似文献
9.
P A di Sant'Agnese K L de Mesy Jensen T A Bonfiglio D E King S F Patten 《Acta cytologica》1985,29(3):477-483
The diagnostic and didactic utility of plastic-embedded semi-thin sections of fine needle aspiration biopsies is presented using a case-study approach. The Spurr epoxy semi-thin sections were stained with a newly developed sequential basic fuchsin-methylene blue stain, which gives hematoxylin-and-eosin-like staining and simultaneously substitutes for a wide variety of special stains. The informational content of the sections can approach that of electron microscopy. The use of a direct off-the-slide "pop-off" technique in preparing the plastic-embedded sections allows for a direct comparison between similar groups of cells embedded in plastic and present on the routine aspiration slides; retrospective analysis can discern subtle, previously unrecognized morphologic features in the alcohol-fixed, Papanicolaou-stained slides. The limitations of this comparative approach, however, become manifest when the effects of alcohol fixation on cells are directly compared in plastic and at the ultrastructural level to aldehyde fixation. 相似文献
10.
Feulgen, Iron-Propionocarmine and Cupra-Ammonium in Preparing Algal Chromosomes for Light Microscopy
Louis A. Hanic 《Biotechnic & histochemistry》1979,54(3):129-133
A method for obtaining algal chromosomal preparations is described employing the Feulgen method for DNA staining, Fe-propionocarmine as an enhancing stain, and cupra-ammonium to remove cell wall material. Fe-propionocarmine applied as a gradient to the slide provides cells stained with the Feulgen stain alum or with the Feulgen Fe-propionocarmine stain, thereby facilitating useful comparison.
Where dilute the Fc-propionocarmine enhances nuclear staining without staining other organelles; where more concentrated it also stains the nucleolus, spindle, spindle polar bodies, pyrenoid and protoplast. Treatment with cupra-ammonium, to remove polysaccharide wall material, followed by neutralization with propionocarmine, enables thinner squashes and better chromosome spreads without IOU of differential staining. Preparations mounted in euparal are long-lasting. 相似文献
Where dilute the Fc-propionocarmine enhances nuclear staining without staining other organelles; where more concentrated it also stains the nucleolus, spindle, spindle polar bodies, pyrenoid and protoplast. Treatment with cupra-ammonium, to remove polysaccharide wall material, followed by neutralization with propionocarmine, enables thinner squashes and better chromosome spreads without IOU of differential staining. Preparations mounted in euparal are long-lasting. 相似文献
11.
J. D. Dodge 《Biotechnic & histochemistry》1964,39(6):381-386
Dinoflagellate chromosomes in sections of plastic-embedded cells were stained without removing the plastic. Azur B and Feulgen procedures were used to localise DNA. Azur B was used with Araldite or methacrylate sections by staining in 0.2% stain in 0.05 M citrate buffer at pH 4 for 1 hr at 50 C followed by rinsing in tertiary butyl alcohol to differentiate the chromosomes. Feulgen stain was used with Araldite sections by hydrolyzing in 1 N HCl at 60 C for 10 min, rinsing in water, staining for 24 hr, washing well, drying and covering. Fast green was used with methacrylate sections to stain proteins by flooding the slide with a 0.1% solution of stain in 0.06 M phosphate buffer at pH 8, allowing the stain to dry out at 40-50 C, washing well, drying and covering. Controls were carried out on material fixed in formalin and treated with nucleases or proteolytic enzymes prior to embedding, and staining. 相似文献
12.
蓟马采集和玻片标本的制作 总被引:18,自引:0,他引:18
介绍蓟马标本的采集方法、常规鉴定用的临时性玻片和存档及分类用的永久性玻片标本的制作方法。标本的采集主要是拍打植物花朵、叶片及枯枝,玻片的制作重点介绍了制作存档和分类用的永久性玻片的5个步骤,即浸解脱色、洗涤、脱水、整姿封盖和干燥。 相似文献
13.
《Biotechnic & histochemistry》2013,88(6):381-386
Dinoflagellate chromosomes in sections of plastic-embedded cells were stained without removing the plastic. Azur B and Feulgen procedures were used to localise DNA. Azur B was used with Araldite or methacrylate sections by staining in 0.2% stain in 0.05 M citrate buffer at pH 4 for 1 hr at 50 C followed by rinsing in tertiary butyl alcohol to differentiate the chromosomes. Feulgen stain was used with Araldite sections by hydrolyzing in 1 N HCl at 60 C for 10 min, rinsing in water, staining for 24 hr, washing well, drying and covering. Fast green was used with methacrylate sections to stain proteins by flooding the slide with a 0.1% solution of stain in 0.06 M phosphate buffer at pH 8, allowing the stain to dry out at 40-50 C, washing well, drying and covering. Controls were carried out on material fixed in formalin and treated with nucleases or proteolytic enzymes prior to embedding, and staining. 相似文献
14.
Kevin Grayson 《Biotechnic & histochemistry》1978,53(2):115-120
This report details a whole mount technique which allows rapid assessment of meiotic stages in large numbers of mammalian oocytes using an aceto-orcein stain.
Previous studies of mammalian oocyte meiosis have been hampered by the use of time-consuming staining techniques. Temporary preparations have been produced using squashes of oocytes with fixative and stain drawn under the coverslip by diffusion (Austin 1961). An air-drying technique requires less attention by the technician and allows permanent preparations to be made (Tarkowski 1966). However, these methods require that each slide be treated individually. The improvements outlined below permit many slides to be processed together, allowing surveys of larger numbers of oocytes than before. 相似文献
Previous studies of mammalian oocyte meiosis have been hampered by the use of time-consuming staining techniques. Temporary preparations have been produced using squashes of oocytes with fixative and stain drawn under the coverslip by diffusion (Austin 1961). An air-drying technique requires less attention by the technician and allows permanent preparations to be made (Tarkowski 1966). However, these methods require that each slide be treated individually. The improvements outlined below permit many slides to be processed together, allowing surveys of larger numbers of oocytes than before. 相似文献
15.
Purified preparations of tobacco mosaic virus (TMV) and pieces of tomato leaves infected with TMV were embedded in methacrylate or epoxy resin, sectioned, and stained with 1.0% strontium permanganate for electron microscopy. In sections containing purified and intracellular virus, the apparent length of stained particles varied directly with section thickness, indicating stain penetration beyond the surface of the section. Penetration was demonstrated also by stereoscopy. Penetration was less complete when sections were allowed to dry before staining. In most instances the number of identifiable particles per unit area was independent of section thickness but increased when both surfaces of the sections were stained instead of only one surface. Staining was prevented by thin films of methacrylate or epoxy resin placed between the virus section and staining solution. Most results supported the view that electron scattering capacity was enhanced only in particles which intersected the surface of the section exposed to permanganate. 相似文献
16.
Preparation of stained smears of Entamoeba histolytica has several drawbacks. We therefore tried to simplify the staining procedures by modifing Kohn's chlorazol black E staining and Wheatley's trichrome staining techniques. Trophozoites and cysts of axenically cultured E. histolytica and Entamoeba invadens, respectively, and trophozoites and cysts of E. histolytica in stools of patients were used. Karyosomes and peripheral chromatin of nuclei and chromatoid bodies became distinctly visible after amoebae were suspended in the basic solution of Kohn's stain. Amoebae fixed in suspension with either basic solution or Bouin's fixative were clearly stained with Kohn's and trichrome preparations, both as wet mounts directly and as permanent slides after processing for mounting. These procedures were easier when the basic solution was used as a fixative and trichrome stain was employed. Erythrocytes ingested by trophozoites, however, were not stained with either of these preparations after fixation in the basic solution but were clearly stained when Bouin's fixative was used. Cysts of E. histolytica in stools concentrated using basic solution (instead of formalin) and ether were also stained with these stains. Consequently, without employing highly toxic mercuric chloride, wet mounts and permanent smears can be prepared with permanent stains, and preserved cysts can be stained after concentration. 相似文献
17.
Rosette Spoerri 《Biotechnic & histochemistry》1948,23(3):133-135
A new staining method has been developed for the study of nerve cells and Nissl granules which combines three basic dyes, cresylecht violet, toluidine blue and thionin. The use of this tri-basic-dye stain results in finished preparations that are critically stained and permanent. Paraffin sections (4 μ sections preferably) are mounted on slides by the starch medium, deparaffinized and stained by the tribasic staining solution. After differentiation in acidified distilled water, sections are dehydrated, returned to stain solution and again dehydrated, then cleared and mounted in Clarite. Various vertebrate material including normal and pathological human tissues have been stained with this triple dye solution. Especially for pathological material, re-immersion of slides in the staining and 80% alcohol solutions before mounting, differentially intensifies the staining reaction. Fixatives used were 10% formalin, 95% alcohol, Bouin and formalin-Bouin (10% formalin followed by Bouin). 相似文献
18.
Margaret E. Long 《Biotechnic & histochemistry》1948,23(2):69-75
A procedure for the differentiation of the mesenchymal derivatives, myofibrillae, reticular and collagenous fibers is presented. Formol-Zenker fixation (5-12 hours) is followed by the washing, iodinization, dehydration and paraffin embedding steps routine for that fixative with the following modifications. Zirkle's butyl alcohol series is used for dehydration and infiltration with paraffin as well as in the alcohol slide series. Embedding paraffin used is Parawax plus 8-10% bayberry wax. Tissue-exposed surface of paraffin block is soaked in water overnight before cutting serial sections at 3-5μ. Sections are mounted using the dilute albumen method, and the slides, thoroughly dried at 37oC. overnight, are left at 60o for 10 minutes to melt the paraffin of the sections. Before staining, the sections are given a preliminary treatment with potassium permanganate and oxalic acid. For reticular staining a 10% silver nitrate bath is succeeded by an ammoniacal silver carbonate solution followed by reduction in 1% neutral formalin, toning in gold chloride and fixing in sodium thiosulphate. Myofibrillae, the sacroplasmic limiting membrane and other sarcous elements are stained by Heidenhain's azocarmine solution, adult tissues at room temperature and fetal tissues at 50 oC. Differentiation in phosphotungstic acid is followed by the staining of collagenous fibers. For adult tissue, light green SF (C.C.) is used and for fetal tissue, fast green FCF (C.C). A discussion of the preparation of ammoniacal silver solutions is included. Both stock and used solutions of ammoniacal silver have been in use by the author for over a period of two years. 相似文献
19.
We describe a fast method for firm attachment of large plastic sections to glass slides with EVA-copolymers, commonly known as hot melt sticks. Solid hot melt sticks dissolve slowly in xylene to form an adhesive gel within 6 hours. Small drops of hot melt gel are applied to the corners of the sections and surrounding slide surface at ambient or elevated temperatures. The gel sticks to both the plastic and the glass slides. The hot melt “corner point method” prevented detachment of sections in staining procedures. As an additional technique, we suggest the use of hot melt adhesive for attaching plastic specimen blocks to wooden blocks or metallic specimen holders. 相似文献
20.
A gelatin film procedure was used to localize proteolytic activity in the lumen of the intestinal ceca of Leucochloridiomorpha constantiae (Trematoda) adults. Slides were coated with a 7·5% gelatin solution and then fixed in 10% neutral buffered formalin (NBF). Cryostat sections of isolated worms or those attached to the bursa of Fabricius of the domestic chick were affixed to the gelatin film. Experimental and control slides were incubated in a humid chamber for 30 min at 37·5 and 4°C, respectively. Slides were again fixed in NBF, and then stained for protein with mercuric bromphenol blue (MBB). In experimental slides, the lumen of the intestinal ceca was lysed and did not stain, whereas worm and host tissue and the gelatin were protein-positive. Control sections stained uniformly positive for protein. In this procedure tissue is retained on the slide and proteolytic activity can be correlated with a tissue site on the same slide. 相似文献