A Quick Embedding Method for Light and Electron Microscopy of Textile Fibers

A quick embedding method employing UV polymerization reactions has been devised for embedding fibers in acrylic and meth-acrylate media. The resultant thin, flat embed-dings are suitable for both light and electron microscopy.     

植物乳杆菌多孔淀粉直接包埋工艺的研究

利用多孔淀粉的吸附特性,将植物乳杆菌包埋于多孔淀粉内,通过测定多孔淀粉对植物乳杆菌的包埋率,研究菌体浓度、多孔淀粉添加量、振荡转速、包埋温度、pH值、时间对包埋率的影响,确定最适包埋条件。结果表明:菌体浓度10~8cfu/mL,多孔淀粉添加量为2%,pH 6.0、20℃,200 r/min振荡处理40 min,在此条件下包埋率为79.5%,对包埋后的菌体进行喷雾干燥试验,其存活率较未包埋的从0.35%提高到29.5%。     

A Quick Embedding Method for Light Microscopy and Image Analysis of Cotton Fibers

A quick embedding method using UV polymerization of methacrylate plastic has been devised for embedding fibers encased in a polyvinyl chloride tube. The resulting embedments are suitable for light microscopy and image analysis.     

Immunohistochemistry on Resin Sections: A Comparison of Resin Embedding Techniques for Small Mucosal Biopsies

We have modified resin embedding methods to provide optimal information from en-doscopic biopsies. Mucosal biopsies were fixed either in buffered formalin and processed for embedding in Araldite or in acetone containing protease inhibitors and embedded in glycol meth-acrylate (GMA). GMA embedding generated an im-munophenotypic profile similar to that obtained in frozen sections while yielding far superior morphology and greater numbers of sections from small biopsies. The phenotypic markers included those for T cells, macrophages, mast cells, eosin-ophils and neutrophils. We have also demonstrated collagens, cell adhesion molecules and integrin molecules. Sections of similar quality were obtained with Araldite but the repertoire of antibodies was restricted to those which can be applied to formalin fixed, paraffin embedded tissues. We suggest that for optimal results, small biopsies to be subjected to immunochemistry are fixed in acetone at -20 C with the inclusion of protease inhibitors and embedded in GUIA with careful temperature control.     

New embedding medium for sectioning undecalcified bone

Methylmethacrylate (MMA) is the most commonly used embedding medium for sectioning undecalcified bone; however, a number of problems exist with its use in a research laboratory. MMA requires a long infiltration time and temperature control, and it reacts with many polymers. We used Kleer Set resin™ as an alternative embedding medium for sectioning undecalcified bone specimens. Fluorochrome labeled bone specimens were sectioned transversely using a ground section technique and longitudinally on a sledge macrotome. The slides were viewed using both transmitted light and epifluorescence microscopy. High quality sections were obtained using Kleer Set resin™ for both sectioning techniques. We have shown that this new embedding medium is simpler, safer, quicker to use and does not interfere with visualization of fluorochromes.     

New embedding medium for sectioning undecalcified bone

Methylmethacrylate (MMA) is the most commonly used embedding medium for sectioning undecalcified bone; however, a number of problems exist with its use in a research laboratory. MMA requires a long infiltration time and temperature control, and it reacts with many polymers. We used Kleer Set resin? as an alternative embedding medium for sectioning undecalcified bone specimens. Fluorochrome labeled bone specimens were sectioned transversely using a ground section technique and longitudinally on a sledge macrotome. The slides were viewed using both transmitted light and epifluorescence microscopy. High quality sections were obtained using Kleer Set resin? for both sectioning techniques. We have shown that this new embedding medium is simpler, safer, quicker to use and does not interfere with visualization of fluorochromes.     

Histological investigation of organisms with hard skeletons: a case study of siliceous sponges

Siliceous and calcareous sponges commonly are treated with acid to remove the spicules prior to embedding and cutting for histological investigations. Histology of spiculated sponge tissue represents a challenging problem in sponge histotechnology. Furthermore, fluorescence in situ hybridization (FISH), a key method for studying sponge-associated microbes, is not possible after acid treatment. For a broad range of siliceous sponge species, we developed and evaluated methods for embedding in paraffin, methylmethacrylate resins, LR White resin and cryomatrix. Different methods for cutting tissue blocks as well as mounting and staining sections also were tested. Our aim was to enable histological investigations and FISH without prior removal of the spicules. To obtain an overview of tissue and skeleton arrangement, we recommend embedding tissue blocks with LR White resin combined with en bloc staining techniques for large specimens with thick and numerous spicules, but paraffin embedding and subsequent staining for whole small specimens. For FISH on siliceous sponges, we recommend Histocryl embedding if the spicule content is high, but paraffin embedding if it is low. Classical histological techniques are used for detailed tissue examinations.     

兔VX2肝癌模型制作技术改良

目的探讨制作兔VX2肝癌模型的新技术、新方法。方法将新西兰兔30只随机等分为3组,分别为嵌插组、改良嵌插组和经皮穿剌组。分别按不同方法建立肝癌模型,于建模后第1、2、3周分别进行CT扫描,评估肿瘤大小。3周后处死动物进行解剖,观察肝脏成瘤及转移情况,比较各组转移率,并行常规病理组织学检查。结果 30只动物建模成功28只,成功率93%,经皮穿剌组有2只肝脏未见种植灶,嵌插组和改良嵌插组全部种植成功。改良嵌插组腹壁转移率显著低于嵌插组和经皮接种组。结论改良嵌插法3周内不会形成接种部位腹壁以及远处的转移,是一种理想的移植性肝癌模型建模方法。     

Embedding Thin Plant Specimens for Oriented Sectioning

Small plant structures such as small primary roots, filamentous mosses and algae are difficult to orient for sectioning since they become wavy and curl during embedding. A method is described for embedding and orienting tiny plant specimens in a glycol methacrylate resin using self-constructed flat molds. Prior to sectioning, small samples can be oriented in both the longitudinal and the transverse plane. As several samples can be sectioned simultaneously, time-consuming trimming of the blocks is reduced substantially. The efficiency of this technique has been demonstrated using the tiny roots of the model plant Arabidopsis thaliana (L.) Heynh.     

In Situ Embedding of Cell Monolayers Cultured on Plastic Surfaces for Electron Microscopy

Various procedures suitable for routine in situ embedding of cell monolayers were tested including: (1) the use of different Epon substitutes, (2) the use of different types of plas-ticware obtained from different sources, and (3) different methods of preparing capsules for sectioning. Different resins reacted differently with different plastics and type of preparation. Merck Epon substitute bound to most of the plastics tested. Ladd Epon substitute released cleanly from all plastics tested when a suitable method of preparation was used. The results show that for routine embedding of cell monolayers it is necessary to select an appropriate Epon substitute and method of preparation of capsules for the type of plasticware used. A routine method is described, with various alternative steps which can be applied when particular difficulties are encountered.     

响应面法优化火麻仁油微胶囊喷雾干燥工艺的研究

为了制备包埋率高、稳定性好的火麻仁油微胶囊,拓展其在食品领域的应用范围,以火麻仁油为芯材、单双脂肪酸甘油酯为乳化剂、酪蛋白酸钠为壁材、固体玉米糖浆为填充剂、柠檬酸钠为缓冲盐、抗坏血酸棕榈酸钠为抗氧化剂,通过喷雾干燥法制备60%载油率的火麻仁油微胶囊,以微胶囊包埋率为响应值,在单因素实验的基础上,以干物浓度、进风温度、出风温度为实验因素,采用Box-Behnken响应面分析法进行优化。随后通过扫描电镜观察火麻仁油微胶囊表面形态结构,以确定包埋效果。并利用油脂氧化分析仪检测火麻仁油微胶囊的氧化稳定性。研究确定微胶囊的最佳工艺条件为：干物浓度42%、进风温度168 ℃、出风温度74 ℃,在此条件下制备得到的火麻仁油微胶囊包埋率可达92.15%。通过扫描电镜观察到火麻仁油微胶囊表面圆滑无裂痕,表明火麻仁油微胶囊包埋效果比较理想。经油脂氧化分析仪测定,与对照组（火麻仁油）相比,试验组（火麻仁油微胶囊）的氧化诱导期时间较长,能够达到30 h以上,说明通过对火麻仁油进行微胶囊包埋可以较大程度地提高油脂的稳定性。研究结果为火麻仁油在食品工业领域的开发和应用提供了理论支持。     

百会穴埋线配合认知行为干预对卒中后便秘患者的影响

目的:探讨百会穴埋线配合认知行为干预对卒中后便秘患者的影响,为减轻临床脑卒中患者便秘症状提供思路。方法:选取黑龙江省某三级甲等医院针灸三科住院的脑卒中便秘患者120例,随机分为四组,对照组给予常规护理,其他组均在常规护理的基础上予以相应的干预措施,试验组1给予认知行为干预,试验组2给予百会穴埋线,试验组3给予百会穴埋线加认知行为干预。比较干预前、干预后7天、干预后14天的四组便秘临床症状积分、便秘治疗效果。结果:干预14天后,四组便秘症状积分均显著低于干预前(P0.05),且试验组1、试验组2和试验组3便秘症状积分均低于对照组,试验组3便秘症状积分最低(P0.05),试验组3的便秘治疗总有效率为92.86%,显著高于其他三组(P0.05)。结论:与常规治疗、常规治疗联合百会穴埋线或认知行为干预相比,百会穴埋线配合认知行为干预可显著提高脑卒中患者便秘的临床效果。     

Histochemical and immunohistochemical protocols for routine biopsies embedded in Lowicryl resin

Tissue processing and analysis require good preservation of both the shape and content of cells. Lowicryl resin is one of the few embedding media that allow good preservation of both tissue architecture and cellular contents. Therefore, different histochemical and immunohistochemical reactions can be applied to semithin sister sections from one biopsy. Further examination of a zone of interest can be carried out under the electron microscope. The hydrophilic property of Lowicryl resins makes possible different histochemical reactions; however, the technique used for paraf?n sections must be adapted for each reaction. Antigenic preservation of cells by low temperature embedding allows immunolabeling on either semithin sections or in the zone of interest on ultrathin sections. We have shown the application and adaptation of different histochemical and immunohistochemical reactions on semithin and ultrathin sections from hepatic biopsies that were large, but thin. The variety of techniques that can be used on sister Lowicryl sections of a single biopsy makes this medium useful for extensive pathological studies of precious needle biopsies.     

The Analysis of the Geometric Structure of the Delayed Time Used in the Embedding Process in Predicting Chaotic Time Series

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A Plastic Embedding Technique for Analyzing Fluorescent Dextran-Amine Labelled Neuronal Profiles

A plastic embedding technique employing fluorescently labelled dextran-amines is described. After application of tracer to cut nerves and appropriate transport time, animals were fixed in paraformaldehyde. Subsequently their brains were dissected, heads and brains were dehydrated, embedded in methacrylate and sectioned serially on a rotary microtome. Plastic sections allow high resolution of single neuron profiles and complete serial reconstruction of un-distorted sections, including embryos with large amounts of yolk. In conjunction with whole mount analysis and double labelling, this technique can accurately reveal the spatial relationships of nerve components throughout development.     

Chemical Dehydration for Rapid Paraffin Embedding

We describe chemical dehydration with 2,2-dimethoxypropane (DMP) for rapid paraffin embedding using a mixture of DMP and mineral oil followed by mineral oil as clearing intermediates. This method is useful for classical histological techniques as well as for histochemistry and immunocytochemistry.     

微贮用植物乳杆菌固定化菌剂的制备

乳酸菌与纤维素降解菌因其可防止微贮饲料酸败、增加秸秆饲料的营养价值等优点,在秸秆微贮过程中起重要作用。但由于乳酸菌的繁殖会抑制纤维素降解菌的活性,如何实现微贮过程中两种微生物分时发挥功能是解决上述问题的关键。文中利用固定化技术将乳酸菌制备成含有玉米秸秆粉的固定化菌剂以达到缓释的目的。首先制作固定化空白小球得出复合固定化载体成球的最佳浓度,利用玉米芯吸附植物乳杆菌S1得到复合固定化载体,以对S1的包埋率、成球效果等为指标,通过对比两种固定化方法 (包埋法与包埋-交联法),得到固定化植物乳杆菌S1的最佳条件。研究表明,使用6%PVA+0.4%SA+0.3%CMC-Na进行包埋-交联时成球效果最好,使用1.2%SA+0.5%CMC-Na进行直接包埋时成球效果最好。通过对比5种固定化工艺,将1.2%SA+0.5%CMC-Na和吸附玉米粉组成的固定化载体混合物逐滴滴入4%氯化钙中直接包埋24 h得到的固定化小球其机械强度以及包埋率均优于其他工艺。因此,利用玉米芯吸附-海藻酸钠包埋的方法可以有效提高植物乳杆菌包埋效率,为使用固定化技术制备微贮饲料菌剂奠定基础。     

Preparation of Thin Sections for Fourier Transform-Infrared Microscopy: a New Embedding Medium for Cellulosic Samples

Fourier Transform-Infrared [FT-IR] microscopy is a combination of instrumentation from which information can be derived about the structure and composition of materials; however, it presents unique problems for sample preparation. Traditional methods of preparing fiber cross sections employ embedding media such as methacrylates, epoxides and polyvinyl alcohols, all of which have groups in common with the cellulose molecule, and absorb in the same regions of the IR spectrum. Therefore, a new embedding method employing polystyrene has been developed for the preparation of cross and longitudinal sections of cellulosic fibers. Although polystyrene is a strong IR absorbing material, it can be completely removed from specimens prior to analysis. In addition, FT-IR spectra of cross sections have better resolution than conventional preparation methods employing ground samples prepared in a KBr disk.     

A RAPID, HIGHLY EFFICIENT METHOD FOR COLLECTING, FIXING, AND EMBEDDING PLANKTONIC AND OTHER SMALL CELLS FOR ELECTRON MICROSCOPY

A rapid, highly efficient method is described for fixation, dehydration, and embedding of small (e.g. planktonic) cells dispersed in large volumes of culture medium. The entire protocol, based on continuous filtration, can be completed within about an hour, and the yield of cells is very high. Fixation quality has been excellent with several different types of samples.     

7 Å projection map of the S-layer protein sbpA obtained with trehalose-embedded monolayer crystals

Two-dimensional crystallization on lipid monolayers is a versatile tool to obtain structural information of proteins by electron microscopy. An inherent problem with this approach is to prepare samples in a way that preserves the crystalline order of the protein array and produces specimens that are sufficiently flat for high-resolution data collection at high tilt angles. As a test specimen to optimize the preparation of lipid monolayer crystals for electron microscopy imaging, we used the S-layer protein sbpA, a protein with potential for designing arrays of both biological and inorganic materials with engineered properties for a variety of nanotechnology applications. Sugar embedding is currently considered the best method to prepare two-dimensional crystals of membrane proteins reconstituted into lipid bilayers. We found that using a loop to transfer lipid monolayer crystals to an electron microscopy grid followed by embedding in trehalose and quick-freezing in liquid ethane also yielded the highest resolution images for sbpA lipid monolayer crystals. Using images of specimens prepared in this way we could calculate a projection map of sbpA at 7 Å resolution, one of the highest resolution projection structures obtained with lipid monolayer crystals to date.