A Staining Rack for Handling Cover-Glass Preparations

A porcelain staining rack (Fig. 1) has been devised for handling cover-glass preparations. The design is along the general lines of staining racks for slides commonly sold by commercial houses. It consists of three parallel rods—one at the bottom, two on the sides. These three rods are held together by two end pieces. Each of the rods has twelve slots, thus the rack holds twelve cover glasses. The slots have been arranged to hold the cover glasses some distance apart. Circular cover glasses having a diameter of 22 mm. are most desirable. Each of the two end pieces has a hole near the top for inserting one end of the wire tongs or handle (see Fig. 1) which is used for removing the rack from the stender dish. The staining rack is 40 mm. long and 35 mm. high and is designed, as shown in the figure, to fit the ordinary Stender dishes commonly used in many laboratories.     

A Plexiglas Isolation-Transfer Chamber for Use with the Fonbrune Micromanipulator

The chamber is made with two pieces of clear Plexiglas, both 1 inch wide and 3 inches long; the top piece, 1/16 inch thick; the bottom, 1/4 inch. A recess 14 mm wide, 37 mm long and 5 mm deep is cut into the bottom piece leaving a margin 1.5 mm wide, this margin is then cut down to a height of 2.5 mm for the length of the recess; a piece of moistened filter paper I1/2 inches long and 5 mm deep is attached to the rear wall, leaving the bottom clear for the transmitted light; two notches 13 × 15 mm and 12 mm apart are cut from the top piece. The top is superimposed on the bottom and held by two screws to form a chamber that is accessible to microdissection instruments through its open side. Two cover glasses, one bearing the specimen and the other, the medium to which the transfer is to be made, are placed over the notches in the top (agar side down). The actual transfer of material is made by shifting the position of the cover glasses with the mechanical stage of the microscope while the specimen is held by the micromanipulator.     

A Plexiglas Isolation-Transfer Chamber for Use with the Fonbrune Micromanipulator

The chamber is made with two pieces of clear Plexiglas, both 1 inch wide and 3 inches long; the top piece, 1/16 inch thick; the bottom, 1/4 inch. A recess 14 mm wide, 37 mm long and 5 mm deep is cut into the bottom piece leaving a margin 1.5 mm wide, this margin is then cut down to a height of 2.5 mm for the length of the recess; a piece of moistened filter paper I1/2 inches long and 5 mm deep is attached to the rear wall, leaving the bottom clear for the transmitted light; two notches 13 × 15 mm and 12 mm apart are cut from the top piece. The top is superimposed on the bottom and held by two screws to form a chamber that is accessible to microdissection instruments through its open side. Two cover glasses, one bearing the specimen and the other, the medium to which the transfer is to be made, are placed over the notches in the top (agar side down). The actual transfer of material is made by shifting the position of the cover glasses with the mechanical stage of the microscope while the specimen is held by the micromanipulator.     

Microscopy with Plastic Substitutes for Cover Glasses

To determine whether plastic substitutes for cover glasses on microscope slides affect the performance of the microscope, their optical constants were determined. The plastic covers and glass cover glasses were mounted also on silvered slides to form Star Test Plates which were studied by competent observers. The thicker plastic cover glasses, now on the market, are satisfactory when mounted to give a plane surface. Optically inhomogeneous materials, of irregular thickness, those that curl or do not have plane surfaces, adversely affect the performance of the microscope and should not be used. Since the substitutes are softer than glass they must be protected from abrasion. It is recommended that thicknesses of 0.18 mm., and none outside of a range of 0.12 to 0.20 mm. be used. For critical observation with unimmersed objectives of high aperture, best results are obtained with the correction collar set at the position corresponding to the actual thickness of the cover slip, just as would be done with glass cover glasses.     

Femoral chordotonal organ in the legs of an insect,Chrysoperla carnea(Neuroptera)

The femoral chordotonal organ (FCO) inChrysoperla carneais situated in the distal part of the femur and consists of two scoloparia, which are fused at their distal end. The distal scoloparium contains 17-20 scolopidia, and the proximal one six scolopidia. Each scolopidium consists of two sensory cells and three types of enveloping cells (glial, scolopale and attachment cell). The sensory cells of different scolopidia do not lie at the same level in the FCO. Therefore the attachment cells of different scolopidia have different lengths. In the FCO, three types of ciliary roots are found in different sensory cells. The dendrite of the sensory cell terminates in a distal process, which has the structure of a modified cilium (9x2+0). The very distal part of the cilium is surrounded by an extracellular electron dense material, the cap, and ends in a terminal dilation. The scolopale cell contains the electron dense scolopale rods, consisting of plentiful microtubules. In their middle third the scolopale rods are fused and form the scolopale. In the FCO septate junctions, desmosomes and hemidesmosomes are found.     

Distribution and morphology of the ampullary organs of the salmontail catfish,Arius graeffei

Whole body staining of Arius graeffei revealed that ampullary pores cover the body with their highest densities occurring on the head and lowest densities on the mid‐ventral surface. Each ampullary organ consists of a long canal (0.2–1.75 mm) passing perpendicular to the basement membrane, through the epidermis into underlying dermal connective tissues, curving thereafter to run roughly parallel to the epidermis. Histochemical staining techniques (Alcian blue and Lillie′s allochrome) indicate that the canals contain a neutral to acidic glycoprotein‐based mucopolysaccharide gel that varies in composition along the length of the canal. Collagen fibers, arranged in a sheath, surround a layer of squamous epithelium that lines each ampullary canal. At the proximal end of the canal, squamous cells are replaced by cuboidal epithelial cells that protrude into the lumen, thus constricting the lumen to form a small pore into the ampulla. The ampulla is lined with receptor and supportive cells. The numerous (60–120) pear‐shaped receptor cells bear microvilli on their luminal surface. Two forms of receptor cells exist in each ampullary organ: basal and equatorial receptor cells. Each receptor cell is connected to an unmyelinated nerve. Each receptor cell is surrounded by supportive cells on all but the apex. Tight junctions and underlying desmosomes occur between adjacent receptor and supportive cells. This form of ampullary organ has not previously been described for teleosts. J. Morphol. 239:97–105, 1999. © 1999 Wiley‐Liss, Inc.     

大黄鱼仔稚鱼脊柱、胸鳍及尾鳍骨骼系统的发育观察

研究采用二重染色法对3-28日龄人工培育的大黄鱼Larimichthys crocea(Richardson)仔稚鱼的部分骨骼系统的发育进行观察。结果显示,仔鱼的脊索从6日龄(平均全长MTL3.9mm)开始分节,8-10日龄髓弓、脉弓先后开始发育,14日龄(MTL7.5mm)脊椎骨、髓棘、脉棘均已形成,至28日龄(MTL19.5mm)已发生骨化;大黄鱼仔鱼孵化时,胸鳍的上匙骨、匙骨、后匙骨、支鳍骨原基已形成,27日龄(MTL19.0mm)稚鱼具5块支鳍骨,鳍条发育完整;尾鳍发育以8日龄仔鱼脊索后端腹部2块尾下骨的形成开始,至14日龄仔鱼尾鳍已初具雏形,尾杆骨、尾上骨和尾下骨均发育完全,21日龄稚鱼尾下骨排列成弧状,第三与第四块尾下骨之间的缝隙增大,把尾鳍鳍条分为上下两部分,鳍条分节明显,28日龄稚鱼尾鳍尾杆骨及鳍条发生钙化。       

A new technique for the surgical management of deformities in the growing spine.

The Luque procedure was developed to correct the deformity without the need of bracing and maintaining that correction with growth. However many authors are disappointed by their results and the complications which appear in the management of infantile scoliosis with Luque trolley alone. Besides failed implants, pseudarthrosis, modest spinal growth and protuberant rods and wires, the major problem of the Luque systems is the high incidence of loss of correction by postoperative rotation. Therefore a new application technique is recommended. A standard posterior extraperiostal approach is chosen. Sublaminar titanium cables are passed at each level except the caudal lamina. Then the rods are precontoured in shape of the planed curve correction. We use a low profile titanium instrumentation with 5.0 mm diameter rods and 4.2 mm pedicle screws. In contrast to the conventional use of two antiparallel "L"-rods we recommend the use of one reversed "U"-rod securing the laminae with sublaminar titanium cables of the upper end vertebrae. For fixation of the lower spine a dual-opening pedicle screw system is used. Using a holding forceps the distal rods are introduced and fixed into the side opening of the screws then secured by sublaminar wires. In addition both single rods are stabilized by a low profile cross link bar. This technique allows to correct pelvic obliquity and a stable anchorage of two screws reduces risk of postoperatively rotation or caudal rod slippage due to gravity forces.     

A Portable Rack for Staining Jars

A simple and inexpensive rack has been designed for the transport and storage of stains and reagents in Coplin staining jars in bacteriological and histological laboratories. The rack promotes ease of carrying, prevents spillage, and keeps the jars permanently in the correct sequence. Details of construction are given. The design can be modified to hold as many jars as necessary and can also be modified to accommodate Stender or other type staining jars or reagent bottles.     

Identification of rod- and cone-specific phosducins in teleost retinas

Phosducin (PD) is a regulatory protein of vertebrate phototransduction cascades. In mammalian retina, it has been thought that only one kind of PD commonly exists in both rods and cones. However, we have found two kinds of PD (OIPD-R and OIPD-C) in the retina of a teleost, medaka (Oryzias latipes). In situ hybridization and immunohistochemical analysis demonstrated that OIPD-R and -C are selectively expressed in rods and cones, respectively. The antiserum against medaka PDs recognized two kinds of proteins in bluegill (Lepomis macrochirus) retina. These results suggest that rod- and cone-specific PDs exist in teleost retinas, probably creating differences in light adaptation between rods and cones.     

Differentiation of wine lactic acid bacteria species based on RFLP analysis of a partial sequence of rpoB gene

PCR-RFLP analysis of rpoB sequences were used to identify lactic acid bacteria (LAB) species commonly isolated from wine. Strains of seven cocci and 12 lactobacilli species could be identified after single digestion with AciI for the cocci and two or three digestions (AciI, HinfI and MseI) for the rods and preceded by colonies isolation on solid selective medium and microscope observation to distinguish cocci and rods cells.     

心室辅助径流泵的设计方法

我们正在研制一种心室辅助径流泵,可在体外循环中代替血泵使用,也可作为心衰病人短期心脏辅助泵使用。此种无密封件的径流的泵的轴尖支承结构由一根据氧化铝陶瓷制成的叶轮轴和两只氧化锆陶瓷制成的轴尖轴承组成。叶轮的外径为７２毫米,进流处直径为２４毫米。位于叶轮上表面的六个叶片的进流侧高度为５．５毫米。出流侧高度为３毫米。     

Studies on polynucleotides. XCVI. Repair replications of short synthetic DNA's as catalyzed by DNA polymerases

Repair replication of short synthetic DNA's corresponding to parts of the gene for the major yeast alanine transfer RNA has been studied. The enzymes used were the Escherichia coli, Microccua luteus and T4 DNA polymerases, similar results being obtained with all the three enzymes. The DNA's used (Fig. 1) were: four double-stranded DNA's with the termini containing the 5′-hydroxyl group protruding by one, three, four or ten nucleotide units and two single-stranded DNA's 29 units long which are capable of folding back on themselves. The aspects inve tigated were: (1) completion of repair, (2) the minimum size of the polynucleotide chains required as primers and those which can serve as templates and (3) the minimum size of the primers which can abolish hairpin formation so as to give the “normal” repair replication. Repair replication of DNA's (DNA-I to DNA-IV: Fig. 1) was characterized to be essentially complete. The minimum size of the primer for repair replication of an extended single-stranded deoxypolynucleotide was ascertained to be about five to seven units long, while the primers required to overcome the hairpin formation in DNA-V and DNA-VI were found to be about twelve units long. Thus, in the presence of a suitable primer, the nucleotide incorporation which occurs in DNA-V in the absence of added primer was completely replaced by incorporation expected for normal repair replication. The minimum size of a chain which can serve as a template was also found in the present work to be about 12 units long.     

紫云英根瘤菌菌株107exo基因簇的遗传互补分析

紫云英根瘤菌菌株１０７经Ｔｎ５插入诱变,得到１２株胞外多糖缺陷型变种,以质粒ｐＭＮ２为载体,从其中７株ＥＰＳ－变种内分别构建了７个Ｒ－Ｐｒｉｍｅ质粒（ｅｘｏＲ＇）,大部分变种的ＥＰＳ－表型可被ｅｘｏＲ＇互补,恢复野生型表型（ＥＰＳ＋）。互补表明,１２株ＥＰＳ－变种可分为６个不同的互补群,其中５个在遗传上连锁。ｅｘｏＲ＇酶切分析,除ｅｘｏＲ＇－０２,ｅｘｏＲ＇－０４外,其余５只的外源片段均整合于ＰＭＮ２的两同向重复序列ＩＳ２１之间。     

广东省发现长指鼠耳蝠及其回声定位声波特征

2013年7月在广东南岭国家级自然保护区阳山县境内(24°48′39.5′′N,112°51′01.3′′E,海拔155 m)捕捉到4只雄性蝙蝠,体型小,前臂长34.1~34.7 mm,颅全长14.3~14.7 mm;体毛浓密,背毛毛基黑色,毛尖灰色,腹毛黑或棕色,毛尖奶油白,靠近肛门处腹毛浅灰色,些许白色;翼膜附着于跖部末端;后足特别延展,长度超过胫骨长之半;鉴定为长指鼠耳蝠[Myotis longipes(Dobson1873)]。同时,基于Cyt b基因序列(1 140 bp)构建的部分鼠耳蝠物种系统进化关系,进一步确认上述标本为长指鼠耳蝠,为广东省翼手目新纪录。该种蝙蝠在我国贵州、广西、重庆有分布记载,但标本和相关资料很少。本文给出了长指鼠耳蝠的外形和头骨特征测量数据,并与印度的标本进行了对比。长指鼠耳蝠的回声定位声波为调频型(FM),主频率为68.2 k Hz;此外,对其分类地位和分布状况进行了讨论。标本保存于广东省生物资源应用研究所。     

Biocorrosion and biodeterioration of antique and medieval glass

Over the past few years we have examined various antique and medieval glasses with regard to general biogenic damage, biopitting (crater erosion), bio‐crusts, and opalescent and white biogenic films. Experiments were carried out on pieces from Roman glass bottles excavated near Abu Tor, Sinai, some pieces of green and blue glass from Cologne Cathedral, some pieces from a little church in Evreux, glass samples from the fortress of the former Dukedom of Delmenhorst near Oldenburg, and some neolithic flint tools from the Negev Desert, Israel. Modern glass from a pigsty (19th century) additionally has been used for laboratory experiments on the attack of glass surfaces by fungi and bacteria. Some of the bacteria used in these experiments were isolated from the ancient pieces of glass. Biopitting with structures very similar to the biopitting of marble and limestone was found on almost all specimens. Lichens were not identified directly, but fungi and algae were observed in the pits as well as under the thin layers exfoliating from the Roman glass bottles. Initial steps of colonization and the potential for heavy‐metal accumulation by the isolated bacteria have been shown in laboratory experiments. A fractal dimension of diffusion‐limited disaggregation (DLD) is suggested as one possible explanation for the characteristic form and structure of the microbially induced and shaped biopitting patterns. A biopitting classification is suggested.     

Wigwamma scenozonion sp.nov. (Prymnesiophyceae) from West Greenland

Wigwamma scenozonion sp. nov. (Prymnesiophyceae) is described on the basis of electron microscopy of shadowcast whole mounts prepared from water samples collected in the vicinity of Godhavn (West Greenland) in July and August 1977. This nanoplanktonic coccolithophorid possesses two smooth flagella and a shorter coiling haptonema. Coccoliths of one type cover the whole cell. Each coccolith is composed of a ring of rod-like crystallites joined end to end and arranged parallel to the edge of the oval coccolith base-plates. A single enlarged crystallite is found on most coccoliths. W. scenozonion is distinguished from the two previously described Wigwamma species by the lack of coccolith superstructures and by having one, rather than two rings of crystallites along the base-plate edge. In addition to the West Greenland specimens a single W. scenozonion cell has been encountered in a water sample from Denmark.     

Insect Embryo Chromosome Techniques

The whole-mount method for studying chromosomes of insect eggs is used; the eggs are caused to adhere to cover glasses, which are handled in racks especially designed for carrying large numbers. A basic and helpful change in the usual technique after fixation and before staining involves extraction of the material 1 hr to overnight with a 1:1 methanol-chloroform mixture to remove plasmal-reactive substances. Either leucobasic fuchsin or sulfonated azure A after acid hydrolysis may be used satisfactorily to stain the chromosomes.     

Lamination of Thermoplastic Sheets as a Means of Mounting Histological Material

Permanent preparations of squashes, whole mounts and stained sections can be made by lamination of thermoplastic sheets. Classical procedures of staining and dehydration for sectioned material were used although dehydration after staining was not required for root tip squashes. Arranging the specimen with the identification label between two pieces of clear Vinylite plastic, 15 mils thick, tightening the preparation in a Photo-Seal Kit electric press and laminating for 3 min gave a finished preparation without the use of glass slides and cover slips. For root tip squashes, the stained tip was placed with a drop of stain between two pieces of plastic, squashed and then laminated. This insured retention of all the tissue which is sometimes lost during mounting processes. Preparations of unstained whole mounts were similarly laminated, with an identification label added between the plastic sheets. Stained sections were placed between two sheets of plastic but the identification label was placed on top of the preparation and a third piece of plastic added. This prevented the label from absorbing excess stain and the increased thickness allowed the slide to be used in a mechanical stage. Well preserved slides 18 mo old indicate that the laminated plastic slide is quite durable. It saves time, reveals good cytological detail and avoids some of the laborious features of other methods. It is a technic that can be used in introductory microtechnic and in the preparation of slides for class use in histology.     

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and ‘roll’ mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card, tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.