扫描电子显微镜植物材料的一种化学干燥技术

扫描电镜新鲜生物材料的制备,一般都要经过干燥处理,其目的主要是使样品不产生收缩畸变,以使样品的形态结构保持在生活状态,如此获得的样品图像才自然真实。现代广为应用的扫描电镜生物样品的干燥方法是临界点干燥(CPD)法。另外还有莰烯干燥法、乙腈干燥法和叔丁醇干燥法等。Jemes(1983)应用化学试剂六甲基二硅胺烷(nexamthyl disilazane,简称 HMDS),分子式[(CH_3)_3Si]_2对昆虫组织进行干燥处理并     

高水分、富含淀粉植物组织的扫描电镜制备技术优化

应用常规高真空扫描电子显微镜观察生物样品必须经过脱水和干燥处理,但无论采用临界点干燥还是冷冻干燥方法,都存在样品表面不同程度失真的问题。植物高水分、富含淀粉组织样品经处理后,容易出现淀粉流失、细胞壁变形等现象,从而造成扫描图像粗糙,无法获得真实的细胞内部结构。本文通过对CO_2临界点干燥、化学固定样品冷冻干燥和新鲜样品冷冻干燥3种扫描电镜样品制备技术中后期制样进行机械断裂和液氮脆断改进,优化出两种植物高水分、富含淀粉组织的扫描电镜样品制备方法:(1)样品首先进行FAA化学固定,经冷冻干燥后用液氮脆断,对断面喷金镀膜和扫描电镜观察。利用该方法所得细胞结构完整,细胞壁整齐,淀粉粒和蛋白轮廓明确,可用于分析淀粉粒和蛋白颗粒在细胞内的分布。(2)新鲜样品直接进行冷冻干燥,经液氮脆断后对断面喷金镀膜和扫描电镜观察。利用该方法所得细胞壁整齐,淀粉粒轮廓更清晰,并且无蛋白颗粒干扰,用于分析淀粉粒在细胞内的分布更加理想。     

Comparison of Preparation Methods of Bacterial Cell-Mineral Aggregates for SEM Imaging and Analysis Using the Model System of Acidovorax sp. BoFeN1

Innovative microanalytical methods are valuable tools in geomicrobiology. They often require the use of dried samples, demanding a challenging sample preparation. Since geomicrobiological samples typically have a strongly heterogeneous composition, choosing a preparation method is not straightforward. We therefore compared how different drying methods (critical point drying, hexamethyldisilazane drying, air drying and freeze drying) influence the structure of bacterial cell-mineral aggregates. Each method proved suitable for a specific purpose, but none was able to completely preserve the sample structure. Additional information was obtained on surface alterations by sputter coating and on preservation of extracellular polymeric substances during resin embedding.     

Differential mechanisms to induce dehydration tolerance by abscisic acid and sucrose in Spathoglottis plicata (Orchidaceae) protocorms

Abscisic acid (ABA) and sucrose are known to induce dehydration tolerance of in vitro plant cells and tissues. The present study reports the presence of different mechanisms by which sucrose and ABA improve dehydration tolerance of Spathoglottis plicata (orchid) protocorms. Orchid protocorms were generated aseptically from seeds on Murashig and Skoog medium, and then treated for 7 d in medium containing 10 mg L^?1 ABA and/or 10% (w/v) sucrose. Dehydration tolerance of protocorms was determined at ～25 °C under various drying conditions at relative humidity from 7 to 93%. The actual rate of water loss (i.e. drying rate) was determined using the rate constant of tissue water loss during drying according to the first‐order kinetics. Drying rate affected dehydration tolerance. ABA treatment reduced drying rate and increased dehydration tolerance of protocorms at all relative humidity values tested. However, when compared on the basis of actual drying rates, there was no difference in dehydration tolerance between control and ABA‐treated protocorms, suggesting that ABA‐induced tolerance was correlated with the drying rate reduction. Sucrose treatment was more effective than ABA treatment for the induction of dehydration tolerance. Interestingly, sucrose only slightly affected drying rate. ABA treatment significantly enhanced the synthesis of dehydrin, whereas sucrose treatment primarily resulted in sucrose accumulation. Sucrose treatment also affected protein turnover during drying, causing a significant decrease in protein content in protocorms. Slow drying promoted the degradation of high molecular weight proteins and enhanced the synthesis of low molecular weight dehydrin. The data suggest that different physiological mechanisms are probably involved in the induction of dehydration tolerance by ABA and sucrose treatment.     

Xeroprotectants for the stabilization of biomaterials

With the advancement of science and technology, it is crucial to have effective preservation methods for the stable long-term storage of biological material (biomaterials). As an alternative to cryopreservation, various techniques have been developed, which are based on the survival mechanism of anhydrobiotic organisms. In this sense, it has been found that the synthesis of xeroprotectants can effectively stabilize biomaterials in a dry state. The most widely studied xeroprotectant is trehalose, which has excellent properties for the stabilization of certain proteins, bacteria, and biological membranes. There have also been attempts to apply trehalose to the stabilization of eukaryotic cells but without conclusive results. Consequently, a xeroprotectant or method that is useful for the stable drying of a particular biomaterial might not necessarily be suitable for another one. This article provides an overview of recent advances in the use of new techniques to stabilize biomaterials and compare xeroprotectants with other more standard methods.     

植物材料快速石蜡制片方法

真空干燥箱已越来越广泛地应用于现代生物学研究领域。该文利用真空干燥箱温度和负压的可控制性能,将固定、脱水、透明和石蜡渗透等过程在真空干燥箱中进行,建立起一套可行的植物组织快速石蜡制片方法。结果显示,真空干燥箱的应用加速了多种试剂的渗透速率,提高了切片质量,达到了优化实验步骤、节省实验时间和减少室内有毒化学气体污染的目的。     

Dehydration Kinetics of Embryonic Axes from Desiccation-sensitive Seeds:An Assessment of Descriptive Models

The response of desiccation-sensitive plant tissues to dehydration is significantly affected by dehydration conditions, particularly the rate of drying. Consequently it is important to be able to quantify drying rate. The aim of the study was to assess two models that have been proposed to describe drying kinetics, and thus to provide a quantification of non-linear drying rates, of embryonic axes excised from recalcitrant seeds. These models are an exponential drying time course, and a modified inverse relationship, respectively. For the six species investigated here the inverse function was generally found to fit drying data better than the exponential function under both rapid and slow drying conditions, and so is recommended. The rate of drying, under the conditions used here, was determined by axis size and possibly the nature of the axis outer coverings, rather than the water activity difference between the tissue and surrounding air.     

Drying techniques for the visualisation of agarose‐based chromatography media by scanning electron microscopy

The drying of chromatography resins prior to scanning electron microscopy is critical to image resolution and hence understanding of the bead structure at sub‐micron level. Achieving suitable drying conditions is especially important with agarose‐based chromatography resins, as over‐drying may cause artefact formation, bead damage and alterations to ultrastructural properties; and under‐drying does not provide sufficient resolution for visualization under SEM. This paper compares and contrasts the effects of two drying techniques, critical point drying and freeze drying, on the morphology of two agarose based resins (MabSelect?/d _w ≈85 µm and Capto? Adhere/d _w ≈75 µm) and provides a complete method for both. The results show that critical point drying provides better drying and subsequently clearer ultrastructural visualization of both resins under SEM. Under this protocol both the polymer fibers (thickness ≈20 nm) and the pore sizes (diameter ≈100 nm) are clearly visible. Freeze drying is shown to cause bead damage to both resins, but to different extents. MabSelect resin encounters extensive bead fragmentation, whilst Capto Adhere resin undergoes partial bead disintegration, corresponding with the greater extent of agarose crosslinking and strength of this resin. While freeze drying appears to be the less favorable option for ultrastructural visualization of chromatography resin, it should be noted that the extent of fracturing caused by the freeze drying process may provide some insight into the mechanical properties of agarose‐based chromatography media.     

初生休眠解除状态和干燥处理对水曲柳种子萌发的影响

为探究初生休眠解除状态和干燥处理对水曲柳种子萌发的影响，本文以初生休眠的成熟水曲柳种子为材料，研究经不同裸层积（暖温10周+低温8周、暖温12周+低温8周、暖温10周+低温10周、暖温12周+低温10周）和干燥处理（干燥、不干燥）的水曲柳种子在适宜温度和较高温条件下的萌发表现。结果表明，初生休眠解除状态不同的水曲柳种子在不同温度下的萌发表现具有相似的规律，种子的萌发会受到干燥处理的影响。不经干燥处理的种子解除休眠越充分，其萌发能力就越强，但层积处理后的种子若经过干燥处理，则解除休眠越充分（尤其是低温时间越长），种子萌发能力下降越多。水曲柳种子次生休眠（热休眠）的诱导受种子初生休眠解除状态的影响较小，但受干燥处理影响较大。干燥处理会降低水曲柳种子的萌发能力，尤其是较高温条件下的萌发能力，初生休眠解除越充分的种子萌发受干燥处理影响越大。生产中如需对解除休眠的种子干燥处理，选择暖温10周+低温8周的层积方法处理种子效果最好。     

Effect of drying on the biological activities of a red microalgal polysaccharide

The red microalga Porphyridium sp. produces a polysaccharide exhibiting a variety of biological activities with potential for medical and cosmetic uses. For this reason, it is important that the drying process, which is the end point of production, should not destroy the natural characteristics of the material. The objective of this study was to evaluate the effect of drying at temperatures ranging from 40 to 140 degrees C on the bioactivities of the polysaccharide. Drying the polysaccharide at temperatures above 90 degrees C caused a significant decline in its biological activities (antiviral and anti-cell proliferation) and reduced elasticity, viscosity, and intrinsic viscosity relative to lyophilized polysaccharide and to the starting product. The relationship between molecular weight and intrinsic viscosity indicated that the polysaccharide takes a rigid coil conformation, which stiffens as a result of drying. FTIR analysis revealed that drying caused both significant conformational alterations in the polymer chains and changes in the interaction between the polysaccharide and the glycoprotein to which it is noncovalently associated. Differential scanning calorimetry analysis of the water adsorbed on the charged groups of the polysaccharide showed that drying at higher temperatures increased the bound water content due to dissociation of the polymer chains. Thus, it is recommended that the polysaccharide be dried in a two-step process in which free water is removed by convection and bound freezing water is removed by lyphophilization.     

A New Drying Method of Plant Specimens for Scanning Electron Microscopy the t-Butyl Alcohol Freeze-Drying Method

The t-butyl alcohol freeze-drying method (lnoue and Osatake 1988) is a simple drying method of biological materials for scanning electron microscopy Fixed specimens were immersed in t-butyl alcohol after dehydration throgh a graded series of ethanol. Specimens in the alcohol were then forzen in a refrigerator. They were placed in the bell jar of a vacuum evaporator and simply evacuated with a rotary pump. The samples were completely dried within 40–60 min after the frozen alcohol was sublimated in the vacuum, when the specimen was examined by scanning electron microscopy (SEM), both surface and intracellular structures were demonstrated in three-dimension without any significant drying artifacts. Careful comparison of the results indicated that the SEM imayes obtained by this method were either superior or equal to those obtained by the critical point drying method.     

Comparison of air-drying methods for evaluating the desiccation tolerance of liquid culture-produced blastospores of Paecilomyces fumosoroseus

Various drying methods were tested to identify a standard procedure for evaluating the desiccation tolerance of liquid culture-produced blastospores of Paecilomyces fumosoroseus. Since our work is focused on optimizing fermentation conditions for the production of P. fumosoroseus, the criteria for selecting a drying method included ease of use, moderate spore survival after drying and limited variation in spore survival. Three air-drying methods were tested: P. fumosoroseus blastospores mixed with silica gel, with sand, or with diatomaceous earth. Humidity controlled drying was used in the diatomaceous earth drying method. Blastospore survivals after drying were 19% (C.V. range, 32 to 45%), 82% (C.V. range, 26 to 43%), and 2% (C.V. range 32 to 50%) for the silica gel, sand, and diatomaceous earth methods, respectively. Blastospores dried using the silica gel and sand methods had been rinsed in 0.7 m polyethylene glycol before drying and rehydrated in the same solution for determination of survival. The variation observed within each method was similar. The silica gel drying method was selected as most appropriate for our studies based on moderate blastospore survival (19%) and ease of use.     

Effect of decompression drying treatment on physical properties of solid foods

This study used a decompression drying instrument to investigate the effects of a drying treatment on the physical properties of solid foods. Commercial tofu was used as a model food and was treated at different temperature and pressure conditions in a drying chamber. Overall, high temperatures resulted in better drying. Additionally, pressure in the chamber influenced the drying conditions of samples. Differences in physical properties, such as food texture, shrinkage, and color were observed among some samples, even with similar moisture content. This was caused by differences in moisture distribution in the food, which seems to have manifested as a thin, dried film on the surfaces of samples. It caused inefficient drying and changes in physical properties. Control of the drying conditions (i.e. pressure and heat supply) has relations with not only physical properties, but also the drying efficiency of solid foods.     

A new angiosperms molecular specimen treatment method for field use

Aims This study aims to identify a more convenient drying method for obtaining molecular specimen of angiosperms in the field than the conventional silica gel drying method. Methods The leaves of Prunus serrulata var. lannesiana and Liriope spicata were dried under temperatures of 150 °C, 80 °C, 40 °C as well as under natural conditions, and by the silica gel drying method, respectively. The DNA extracts of various specimens were then analyzed using techniques of spectrophotometer detection, electrophoresis and PCR to evaluate the impacts of different drying treatments to the genomic DNA of testing plants. Important findings The concentrations of total DNA were higher for the specimens dried at 40 °C treatment and by the silica gel drying method than other treatments when assessed by the techniques of spectrophotometer detection and electrophoresis. The concentration of PCR products was highest in the specimens dried at 40 °C. Based on the results, the 40 °C drying can be recommended for obtaining molecular specimens of angiosperms because of its minimum degree of degradation, for convenience of operation and avoiding carrying large amounts of silica gel in field investigations.     

金针菇不同萃取物对肿瘤细胞的影响

比较冷冻干燥、鼓风干燥两种不同干燥方式对金针菇挥发性成分的影响,对干燥后香味物质较丰富的鼓风干燥方式获得的金针菇样品进行不同极性溶剂萃取并进行抗氧化及体外抗肿瘤试验。结果表明：两种干燥方式得到的主要挥发性成分均为醇类和醛类,但鼓风干燥后的金针菇样品中两类物质含量明显高于真空冷冻干燥方式获得的。体外药理实验表明不同萃取物在抗氧化和抗肿瘤细胞中均表现出良好的活性。     

A Simple Drying Method for Field Emission Scanning Electron Microscopy for Chromosomes

Hexamethyldisilazane treatment and subsequent air drying of spread plant chromosomes is compared with critical point drying. The two procedures are equivalent for preparing chromosomes for examination by field emission scanning electron microscopy at low voltage.     

Effects of Periodical Soil Drying and Leaf Water Potential on the Sensitivity of Stomatal Response to Xylem ABA

The study on the changes of stomatal sensitivity in relation to xylem ABA during periodical soil drying and the effect of leaf water status on the stomatal sensitivity has confirmed that xylem ABA concentration is a good indicator of soil water status around roots and the relation between xylem ABA concentration and predawn leaf water potential remained constant during the three consecutive soil drying cycles based on the slopes of the fitted lines. The sensitivity of stomata to xylem ABA increased substantially as the soil drying cycles progressed, and the xylem ABA concentration needed to cause a 50% decrease of stomatal conductance was as low as 550 mnoL/L in the next two soil drying cycle, as compared with the 750 nmol/L ABA in the first cycle of soil drying. The results using the split-root system showed that leaf water deficit significantly enhanced the stomatal response to xylem ABA and the xylem ABA concentration needed to cause a 50% decrease in stomatal conductance was 2 to 4 times smaller in the whole-root-drying treatment than those in the semi-root- drying treatment. These results suggested that the sensitivity of stomata to xylem ABA concentration is not a fixed characteristic.     

Antibiotic resistance and survival of faecal coliforms in activated sludge system in a semi-arid region (Beni Mellal,Morocco)

The activated sludge process is one of the biological treatment methods used in many countries to reduce the high levels of organic and mineral pollutants and pathogenic micro-organisms present in wastewater. The present work was undertaken to study the dynamic and antibiotic-resistance of faecal coliforms (FC) in the activated sludge system of Beni Mellal. This work has also as objective the study of the survival of FC, protozoan cysts, helminth eggs and FC antibiotic resistance in the sludge dehydrated in drying beds in order to know if the agricultural usage of sludge presents any problems to public health. The activated sludge treatment of Beni Mellal resulted in an average reduction of FC and faecal streptococci of 90.75 and 91.06%, respectively. The overall resistance (resistance to at least one antibiotic) of 111 FC strains isolated from the system was 72.07%. This treatment system did not increase the incidence of FC antibiotic resistance in treated wastewaters. The antibiotic resistance of FC was found to be similar in both raw (71.05%) and treated sewage (77.77%). High levels of antibiotic resistance were towards streptomycin (54.05%), ampicillin (42.34%), amoxicillin (42.34%) and amoxicillin–clavulanic acid (31.53%). The treatment of sludge in drying beds appeared to be efficient in eliminating pathogenic micro-organisms: FC, protozoan cysts and helminth eggs. Moreover, the FC antibiotic resistance did not change over time in sludge-drying bed. According to the standard norms, agricultural utilization of this sludge cannot be excluded. However, it is important to study in the receptor environment the survival and the behaviour of antibiotic-resistant FC present in sludge and water.     

周期性土壤干旱和叶片水势对气孔响应木质部ABA灵敏度的影响

研究了周期性土壤干旱期间气孔对木质部ＡＢＡ响应的灵敏度的变化以及叶片水势对灵敏度的影响。实验结果证明了木质部ＡＢＡ浓度是反映根系周围土壤水分状况的一个指标的结论。土壤周期性干旱不影响木质部ＡＢＡ浓度对土壤水分状况的依赖关系,但显著地提高了气孔对木质部ＡＢＡ 响应的灵敏度。根据对实测数据的数学模拟结果显示,引起气孔导度下降５０％ 所需的木质部ＡＢＡ浓度从第一轮土壤干旱的７５０ ｎｍｏｌ／Ｌ降至第二轮土壤干旱的５５０ ｎｍｏｌ／Ｌ。分根实验的结果表明,叶片水分亏缺显著提高了气孔对木质部ＡＢＡ 的响应的灵敏程度,全根干旱中引起气孔导度下降５０ ％ 所需的木质部ＡＢＡ 浓度比半根干旱的小２ ～４ 倍。这表明,气孔对木质部ＡＢＡ响应的灵敏度不是一个固定的特性,可随植物生长环境及许多其他因素的变化而表现出很大的差异     

花菇的冷冻干燥技术研究

实验研究用板层导热法研究了花菇的冻干特性,获得了新鲜花菇的冻干曲线,分析了花菇冻干过程,测定和比较了新鲜花菇和冻干花菇的营养成份。证实试验机的适应性并确定了花菇的冻干工艺,为工业生产提供了理论依据和参考价值。