A method for preservation of soft tissues of marine teleosts for scanning electron microscopy

Soft, internal tissues of marine teleosts have been preserved for scanning electron microscopy in 2% glutaraldehyde in half-strength flounder saline; the osmolality of this fixative approximated the plasma osmolality of marine teleosts. Fixation was followed by washing, dehydration and Freon critical point drying. This procedure has revealed surface features of renal tubules of Anoplogaster and swimbladder of Melanonus     

A new method of preparing insects for scanning electron microscopy.

Acidified 2,2-dimethoxypropane (DMP), used as an alternative to regular fixation and dehydration methods for insects, was found to be the only successful means of preparing the sugarbeet root maggot larva, Tetanops myopaeformis (R?der) (Diptera:Otitidae), for the scanning electron microscope. No morphological changes were evident when DMP treated sugarbeet root maggot adults were compared to fresh (unfixed) adults and glutaraldehyde-osmium tetroxide fixed adults. The method has been used with success on several gropus of insects. Acidified CMP is quickly hydrolyzed by water in tissue to acetone and methanol. DMP is advantageous in that it penetrates water impermeable cuticles rapidly and saves several steps and time in the fixation and dehydration process.     

A new method of preparing fish chromosomes for scanning electron microscopy

A preparative ion-etching technique has been developed which enhances the images of fish chromosomes obtained by scanning electron microscopy.     

A simplified method for the preparation of rotifers for transmission and scanning electron microscopy

Tor TEM and SEM prepatations, rotifers are placed in little panels made of plastic Beem capsules normally used for embedding, with their conical parts cut off and closed by plankton filter cloth. Thus, the risk of losing animals is considerably reduced.     

A new method for isolating pollen and spores from acetate peels for scanning electron microscopy

A procedure for precisely extracting pollen and spores from cellulose acetate peels made from Carboniferous permineralizations is described. This technique produces either whole or sectioned clean grains and allows for the correlation of morphological and ultrastructural features by scanning electron microscopy. The critical examination of pollen and spores from peels prepared for earlier studies is now possible using this technique.     

A simplified method for preparing rotifer trophi for scanning electron microscopy

A simple, quick technique for the preparation of rotifer trophi for scanning electron microscopy is described. The method permits visual monitoring of the extraction process and does not require critical point drying of the specimens. Micrographs showing fine, structural detail of the hard parts of the mastax of representatives of the following genera are presented:Asplanchna, Conochilus, Filinia, Hexarthra, Keratella, Proalides, Synchaeta, andTrichocerca.     

A method to obtain surface strains of soft tissues using a laser scanning device

A three-dimensional laser scanning device was developed allowing surface digitization of musculoskeletal and soft tissue structures under different loads. Image-processing algorithms were formulated for image registration. These were used to determine displacement mapping and then surface strains. Various validation experiments were performed. Accuracy was obtained on a test cylinder after rigid rotation and on a silicon cylinder compressed in four loading steps. The system accuracy (including the scanning and the data evaluation) was +/-0.10% strain in vertical and +/-0.16% strain in shear and circumferential direction for the rigid rotation exhibiting the zero-strain situation. Silicon cylinder compression showed that the accuracy was best for small strains, whereas strains >5% evoked a slight underestimation increasing further with higher strains (error of 0.54% for 7.22% vertical strain). It was possible to increase the accuracy by performing the strain measurements via sub-steps. This had a remaining error of 0.41% for 7.22% vertical strain. A further experiment was carried out in order to acquire the surface strain of a human lumbar intervertebral disc while it was forced to flexion and extension. This study introduced a laser-based scanning method to obtain soft tissue surface strains. It is important to know the strain distribution of musculoskeletal structures and soft tissues. This could help to better understand the mechanical loading of biological structures e.g. the processes in fracture healing. These data could also be used to assist in the validation process for finite-element models.     

Rapid preparation of uncoated biological specimens for scanning electron microscopy.

We have developed a relatively rapid glutaraldehyde-tannic acid (GTA) and osmium tetroxide (OsO4) fixation procedure which permits many types of uncoated biological specimens to be examined in the scanning electron microscope (SEM) at 20 kV without the occurrence of charging. Most specimens taken one day can be examined in the SEM the following afternoon. Types of specimens successfully treated were perfused adult and embryonic rat tissues, confluent human skin fibroblast tissue cultures, plant roots, flowers, seeds, some garden insects, and microcolonies of salivary streptococci. Cells in suspension and extracted human teeth did become electron conductive when treated with the GTA procedure. Most suspended cells must be centrifuged between each solution and the GTA procedure increases the preparation time for these cells. Extracted teeth are usually simply dried and coated. Therefore, the usual SEM preparation techniques are shorter and perhaps more useful for these types of specimens.     

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。     

A new method for predicting the opening angle for soft tissues

The purpose of this paper is to present a simple new method for calculating the opening angle produced by a given residual stress field in a soft biological tissue. The method uses minimization of potential energy, and is therefore named the MPE method. The accuracy of the MPE method is evaluated by comparing the opening angle it predicts to results from a finite element model of the opening angle experiment. We show that the MPE method provides good predictions of the opening angle, and that it is significantly more accurate than two other methods previously used in the literature.     

A method of comparing spermatozoa with light and scanning electron microscopy

A new method of comparing light microscopy and scanning electron microscopy in the study of small cells, such as spermatozoa, that must be examined under oil immersion is described. A grid is etched on the corner of a microscope glass slide, and its inner edges are incised. Its surface area is calculated as a function f the chamber of the critical-point drying apparatus. This method dispenses with the need for any special coverslip and enables the cells to be observed under oil immersion.