鱼杯体虫(Apiosoma piscicola)的光镜及透射电镜观察

对洪泽湖地区鱼杯体虫(Apiosoma piscicola)的活体、固定染色标本形态及其超微结构进行了较为系统地观察、描述,发现虫体大小在南北地区存在显著差异并就这种现象产生的原因进行了讨论,认为这可能是由于南北地区在气候条件、水体生境等各个方面均存在着很大不同,相同祖先种在进行地域辐射时,对各自生存环境长期适应所产生的结果。在超微结构中对虫体表膜、口围纤毛、口围唇、漏斗、横纤毛带及内部胞器进行了仔细的观察,发现杯体虫体内尤其是口区具有很多细菌和有机颗粒,这就证明了其营养是来自于外界水环境而并非宿主,并从杯体虫食物来源的角度论证了杯体虫是一种体外共生体(ectocommensal)而非体外寄生体(ectoparasite)。另外,还观察到虫体尤其是口围唇部的表膜具有非常明显的褶皱,显示了其口区表膜极强的伸缩能力,这也是虫体在受到刺激或形成游泳体(telotroch或swarmer)时能将整个口围盘缩进体内的原因。       

一种淡水缘毛类纤毛虫————沟钟虫的形态学研究

利用活体观察及蛋白银染色技术对一淡水缘毛目纤毛虫——沟钟虫（Vorticella convallaria）的形态学和表膜下纤维系进行了研究。结果表明：沟钟虫的活体个员自然伸展时外形较稳定,并呈明显的倒置钟状,长宽约为50—85μm×40—75μm;口围缘完全外展时为虫体最宽处;伸缩泡一个,较大,位于口围唇下方及口前庭的左侧。胞质均匀而透明,无脂肪滴存在,游泳体呈圆柱形。细胞表面横纹从口围唇到反口纤毛环为74—78条,自反口纤毛环到帚胚为18—24条。大核呈大幅度盘绕的长肠状,两端高度弯曲,纵贯于细胞内。蛋白银制片后表膜下纤维系特征为：虫体纵向纤维稀疏而粗壮,37—42条,似灯笼状;口围盘纤维呈典型的倾斜态分布,并呈放射状排列。第三咽膜（P3）在近口末端处呈明显的分离态,可视为该物种重要的分类学依据。     

蟹累枝虫的光镜及电镜观察

利用活体观察、蛋白银染色及电镜技术对蟹累枝虫(Epistylis eriocheiri)的形态学、表膜下纤维及超微结构进行了较为系统的描述,研究发现:(1)光镜下该种群体双叉分支,活体时虫体表膜柔软,完全伸展时呈长筒状;大核马蹄形,呈横位;伸缩泡单个,斜卧口围缘下方。口围缘纤维上连细密的环状纤维,下接两端较为粗壮,中间细长的纵长纤维,构成了连续的表膜下纤维系。(2)电镜下该种表面具沟、嵴、横纹结构,沟嵴相互交错;口围纤毛花瓣状;虫体表膜层呈齿状,具嵴状突起、泡间微管、表膜孔,胞质层包括内含胞器单一的致密原生质层和富含胞器的疏松原生质层。研究对该种的上述特征在虫体的收缩机制及与其他相似种之间的关系等方面进行了讨论。       

钩刺目纤毛虫--多核前裂虫的超微结构研究

对钩刺目－土壤纤毛虫－多核前裂虫的超微结构做了透射电镜观察。该种表现了与其他钩刺目种类相似的特征,包括体表分布的粘泡以及扁平的表膜泡,其主要细胞结构特征为：（１）－一层连续的支状内质网将虫体细胞质分成内外两部分;（２）线粒体仅密集于内层细胞质;（３）平行唇突表面的特殊的前列横微管敕纱由胞口发出,沿唇突表层向体后发出并纵行环绕于胞咽外侧,此前列横微管束明显地呈片层状构造;（４）射出体包括位于口区的两     

棘腹蛙血液内利川锥虫生活史的研究

利川锥虫（ＴｒｙｐａｎｏｓｏｍａｌｉｃｈｕａｎｅｎｓｉｓＬｉ）寄生在棘腹蛙血液内。虫体波动膜明显。鞭毛较粗,一般不伸出体外形成游离的鞭毛。感染试验表明其中间宿主是绿蛙蛭（ＢａｔｒａｃｏｂｄｅｌｌａｐａｌｕｄｏｓａＣａｒｅｎａ）。虫体在绿蛙蛭嗉囊的上皮细胞内进行裂配生殖或假包囊,产生许多无鞭体、前鞭体和锥虫体等不同发育时期。前鞭体的超微结构特点是：虫体表膜较厚,表膜下微管管壁较厚,线粒体较多,卵圆或长椭圆形,隆嵴不明显。未见高尔基体,内质网很少,不易观察到。胞质中有３—５颗或更多、多角形至圆形,较大的色素体。     

贻贝棘尾虫口器的结构

利用半薄连续切片和超薄切片对贻尾虫口器的显微及亚显微结构进行了观察,澄清了口侧膜和口内膜的立体位置,它们均为纤毛膜,口侧膜发自内唇的边缘,由两排纤毛构成,口内膜发自口围右壁,紧贴口围顶壁向左伸展,由单排纤毛组成,在口内膜的腹侧发现一非纤毛厚膜,暂定名为吞噬辅助膜,它由两层质膜夹一纵行微管束层构成,至口围后部口围带内收之处,此膜再向腹侧延伸出第二个膜,共同构成括号状,至胞口处此膜与口内和口侧膜互相包卷而成一包卷体,共同进入胸咽,经分析认为此膜及其与两纤毛膜形成的包者体,对棘尾虫摄取的大固体食物起重要作用。     

广东肝血簇虫在宿主内脏裂体生殖时期超微结构的研究

本文详细描述了广东肝血簇虫(Hepatozoon guangdongensis)在实验宿主、中国水蛇肺部裂体生殖整个发育过程各期虫体的超微结构。成熟裂殖体内的裂殖子与肺部毛细血管内皮细胞内的裂殖子的超微结构是相似的。裂殖子(3.4×1.3μm)外被由外膜和内膜构成的表膜,它与球虫一样具有包括类锥体在内的完全顶复结构。内皮细胞内的裂殖子和滋养体都没有围虫泡和围虫泡膜包绕。具有内膜的长形滋养体变圆,并外被由宿主细胞产生的围虫泡和围虫泡膜包绕,转变成为圆形的幼期裂殖体,然后发育成为成熟的裂殖体。成熟裂殖体(23×10μm)内含30—50个裂殖子。裂殖体内没有观察到残余体存在。     

镰游仆虫腹面皮层细胞骨架的扫描电镜观察

应用非离子去垢剂抽提和扫描电镜样品制备、观察相结合的方法，显示了镰游仆虫的腹面皮层细胞骨架，详细描述了处于毛基体和毛基体下水平的口围带、口侧膜、额腹横棘毛骨架，以及口围带小膜托架、口侧膜托架、额腹横棘毛托架的主要附属纤维和非纤毛区表膜下皮层骨架的立体图形。作者据所述各种纤毛器托架附属纤维的定位和分布特征推测，这些附属结构可能与细胞内各种纤毛器间的联系，以及包括纤毛器运动在内的整个细胞运动的协调等有关。     

一种游仆虫皮层纤维结构的扫描电镜研究

应用扫描电镜研究了一种游仆虫皮层纤维结构。结果显示,口围带托架由横向组排在一起的小膜托架单元组成,每个小膜托架是由40-50 nm直径的纤维编织成的长方形薄片;波动膜托架是横向平行排列和纵向平行排列两部分共60多股纤维交错编织成的网状结构;额腹横棘毛区表膜下含有纵纤维层、球形纤维层和深部纤维层三层纤维,其纵纤维层在相应于游仆虫皮层脊（或脊和唇）之间的表膜下方被纵沟分成几部分;背面表膜下含有纵纤维,这层纤维下似乎也有球形纤维层。此外,作者也推测了这些皮层纤维结构在游仆虫皮层形态的保持、支持纤毛器和纤毛器运动及形态发生等方面的作用。     

多泡肠袋虫超微结构的研究

对寄生于鲴亚科鱼类肠道中的多泡肠袋虫的超微结构进行了研究,描述了其体表皮层、胞口、核与胞器及腹中凹板各部分的精细构造。结果显示其体表及口区皮层均由表膜和表膜下纤维系统两部分组成;"V"形胞口密被纤毛、对称排布,其咽微丝较体纤毛处更为发达。大核内具多个核仁,异染色质散布核质内;小核内染色质则呈均匀致密分布。另在腹中凹板内质中发现大量支链淀粉粒,并由不连续的微管束沿凹陷边缘将此区域包围起来。同时,对皮层组分和腹中凹板进行了相近物种间的比较分析并对其功能进行了讨论。     

Status of THz-to-Visible Nanospectroscopy Development

Quite unexpectedly, THz and infraredspectroscopy has now a real chance to solveproblems in the nanosciences. This rests ona new microscope technique that overcomesthe Abbe diffraction limit, by using thenear field of a metal antenna in closeproximity to a scanned sample surface. HereI briefly summarize present activities inthe microwave, mid-infrared and visiblespectral ranges. It seems straightforwardand highly desirable to fill the existinggap between about 20 GHz and 20 THz, andattain spatial resolution of 10 nm andbelow also in this important part of theelectromagnetic spectrum.     

Fluorescence Microscopy Gets Faster and Clearer: Roles of Photochemistry and Selective Illumination

Significant advances in fluorescence microscopy tend be a balance between two competing qualities wherein improvements in resolution and low light detection are typically accompanied by losses in acquisition rate and signal-to-noise, respectively. These trade-offs are becoming less of a barrier to biomedical research as recent advances in optoelectronic microscopy and developments in fluorophore chemistry have enabled scientists to see beyond the diffraction barrier, image deeper into live specimens, and acquire images at unprecedented speed. Selective plane illumination microscopy has provided significant gains in the spatial and temporal acquisition of fluorescence specimens several mm in thickness. With commercial systems now available, this method promises to expand on recent advances in 2-photon deep-tissue imaging with improved speed and reduced photobleaching compared to laser scanning confocal microscopy. Superresolution microscopes are also available in several modalities and can be coupled with selective plane illumination techniques. The combination of methods to increase resolution, acquisition speed, and depth of collection are now being married to common microscope systems, enabling scientists to make significant advances in live cell and in situ imaging in real time. We show that light sheet microscopy provides significant advantages for imaging live zebrafish embryos compared to laser scanning confocal microscopy.     

Virtual slit scanning microscopy

We present a novel slit scanning confocal microscope with a CCD camera image sensor and a virtual slit aperture for descanning that can be adjusted during post-processing. A very efficient data structure and mathematical criteria for aligning the virtual aperture guarantee the ease of use. We further introduce a method to reduce the anisotropic lateral resolution of slit scanning microscopes. System performance is evaluated against a spinning disk confocal microscope on identical specimens. The virtual slit scanning microscope works as the spinning disk type and outperforms on thick specimens.     

Perfectly registered multiphoton and reflectance confocal video rate imaging of in vivo human skin

We present a multimodal in vivo skin imaging instrument that is capable of simultaneously acquiring multiphoton and reflectance confocal images at up to 27 frames per second with 256 × 256 pixel resolution without the use of exogenous contrast agents. A single femtosecond laser excitation source is used for all channels ensuring perfect image registration between the two‐photon fluorescence (TPF), second harmonic generation (SHG), and reflectance confocal microscopy (RCM) images. Images and videos acquired with the system show that the three imaging channels provide complementary information in in vivo human skin measurements. In the epidermis, cell boundaries are clearly seen in the RCM channel, while cytoplasm is better seen in the TPF imaging channel, whereas in the dermis, SHG and TPF channels show collagen bundles and elastin fibers, respectively. The demonstrated fast imaging speed and multimodal imaging capabilities of this MPM/RCM instrument are essential features for future clinical application of this technique. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

A developed temporal focusing‐based multiphoton excitation microscope (TFMPEM) has a digital micromirror device (DMD) which is adopted not only as a blazed grating for light spatial dispersion but also for patterned illumination simultaneously. Herein, the TFMPEM has been extended to implement spatially modulated illumination at structured frequency and orientation to increase the beam coverage at the back‐focal aperture of the objective lens. The axial excitation confinement (AEC) of TFMPEM can be condensed from 3.0 μm to 1.5 μm for a 50 % improvement. By using the TFMPEM with HiLo technique as two structured illuminations at the same spatial frequency but different orientation, reconstructed biotissue images according to the condensed AEC structured illumination are shown obviously superior in contrast and better scattering suppression. Picture : TPEF images of the eosin‐stained mouse cerebellar cortex by conventional TFMPEM (left), and the TFMPEM with HiLo technique as 1.09 μm^?1 spatially modulated illumination at 90° (center) and 0° (right) orientations.

     

Portable optical‐resolution photoacoustic microscopy for volumetric imaging of multiscale organisms

Photoacoustic microscopy (PAM) provides a fundamentally new tool for a broad range of studies of biological structures and functions. However, the use of PAM has been largely limited to small vertebrates due to the large size/weight and the inconvenience of the equipment. Here, we describe a portable optical‐resolution photoacoustic microscopy (pORPAM) system for 3‐dimensional (3D) imaging of small‐to‐large rodents and humans with a high spatiotemporal resolution and a large field of view. We show extensive applications of pORPAM to multiscale animals including mice and rabbits. In addition, we image the 3D vascular networks of human lips, and demonstrate the feasibility of pORPAM to observe the recovery process of oral ulcer and cancer‐associated capillary loops in human oral cavities. This technology is promising for broad biomedical studies from fundamental biology to clinical diseases.      

Microscopy basics and the study of actin–actin-binding protein interactions

Actin is a multifunctional eukaryotic protein with a globular monomer form that polymerizes into a thin, linear microfilament in cells. Through interactions with various actin-binding proteins (ABPs), actin plays an active role in many cellular processes, such as cell motility and structure. Microscopy techniques are powerful tools for determining the role and mechanism of actin–ABP interactions in these processes. In this article, we describe the basic concepts of fluorescent speckle microscopy, total internal reflection fluorescence microscopy, atomic force microscopy, and cryoelectron microscopy and review recent studies that utilize these techniques to visualize the binding of actin with ABPs.     

The Structure and Morphogenesis of the Ventral Ciliature in Paraurostyla hymenophora*

SYNOPSIS. The structure and morphogenesis of the ventral ciliature of Paraurostyla hymenophora (Stokes) are described. The oral primordium apparently originates in association with transverse cirrus #6, from which it migrates anteriorly simultaneous with kinetosomal proliferation. The primordium eventually forms an elongate ciliary field from which the future opisthe's fronto-ventro-transverse (FVT) and undulating membrane primordial fields arise. Concomitantly, the future proter's FVT primordial field is initiated by the disaggregation of frontal cirri #4, #5, and #6. Primordia then develop simultaneously within marginal and ventral cirral rows by a disaggregation of cirri within the respective rows, and do not give rise to new cirri until the FVT fields complete segregation into discrete cirri. Near the completion of cirral production from the FVT primordia, each ventral cirral primordium (VCP) forms the 2 rightmost transverse cirri. Segregation of new cirri within the marginal cirral primordia and VCP then occurs, eventually replacing all old cirri within their respective marginal and ventral cirral rows. At the end of cortical morphogenesis, all old ciliary organelles, with the exception of the adoral zone of membranelles, are either reorganized or replaced. These results suggest an evolutionary affinity between the ventral and marginal cirral rows and raise questions about the control of the developmental competence of individual primordia.