Microstructure and Ultrastructure Alterations in the Pallium of Immature Mice Exposed to Cadmium.

The aim of this study was to investigate microstructure and ultrastructure alterations in the pallium of immature mice exposed to cadmium. Forty immature mice were randomly divided into control, 1/100 LD50 (1.87 mg/kg, low), 1/50 LD50 (3.74 mg/kg, medium), and 1/25 LD50 (7.48 mg/kg, high) dose groups. After oral cadmium exposure for 40 days, the pallium of mice was obtained for microstructure and ultrastructure studies. The results showed that both microstructure and ultrastructure alterations of the pallium were observed in all treated mice and the most obvious alterations were in the high dose group. Microstructural analysis showed seriously congested capillary in the pia mater of the pallium in the high cadmium group. Meanwhile, vacuolar degenerate or karyopyknosis presented in some neurocytes, capillary quantity, and the number of apoptotic cells increased, some neurocytes became hypertrophy, the pia mater separated from the cortex, and local hemorrhage and accompanied inflammatory cell infiltration were also observed. Ultrastructural analysis showed that rough endoplasmic reticulum was expanded, heterochromatin marginalized, perinuclear space distinctly broadened, swelling and vacuolization mitochondria appeared, synapse was swelling, presynaptic and postsynaptic membranes presented fusion, and most of mitochondrial cristae were ambiguous. The results indicated that cadmium exposure for 40 days induced dose-dependent microstructure and ultrastructure alterations in pallium of immature mice.

Effect of cadmium exposure on the histopathology of cerebral cortex in juvenile mice.

Cadmium, a heavy metal, is a toxic environmental and industrial pollutant. Exposure to cadmium can lead to the toxic effects in a variety of tissues, also including the brain. The present study investigated the effect of cadmium exposure on the histopathology of cerebral cortex in juvenile mice. Juvenile mice were randomly divided into control, low (1.87 mg/kg), medium (3.74 mg/kg), and high (7.48 mg/kg) dose groups. After cadmium exposure by drinking water for 10 days, the cerebral cortex was obtained for histopathology studies. The medium and high dose of cadmium, rather than low dose, could induce the histopathology alterations of cerebral cortex in a dose-dependent manner. In the high-dose group, microstructure significantly showed pia mater encephali divorcing from cerebral cortex layer, serious hyperemia of blood capillary in pia mater encephali and cerebral cortex, broadening vessel peripheral clearance, a large number of eosinophil leukocyte infiltrating around blood vessel, vacuolar degeneration in part granule cells, and obviously increasing apoptotic cells. Ultrastructure obviously displayed marginalized heterochromatin, incomplete or fused nuclear membranes, broadened perinuclear space, ambiguous mitochondria cristae, decreased synaptic cleft, and fused presynaptic and postsynaptic membrane. Our results revealed that cadmium at the middle and high dose could induce obvious microstructure and ultrastructure alterations of cerebral cortex in juvenile mice, which may be one important mechanism of cadmium neurotoxicity.

High-power top-hat pulses from a Yb master oscillator power amplifier for efficient optical parametric amplifier pumping.

We demonstrate shaping of high-energy broadband Yb amplifier pulses for the generation of a (sub)picosecond top-hat temporal pulse profile that significantly improves pumping efficiency of an optical parametric amplifier (OPA). Phase-only modulation is applied by an acousto-optic programmable dispersion filter. This simple scheme is scalable to a high average power due to a relatively broad bandwidth of the Yb:CaF(2) gain medium used in the amplifier that supports a sub-150-fs transform-limited pulse duration. Additionally we show that OPA seeding with supercontinuum remains possible because top-hat-shaped pulses passed through a glass block recompress to ≈200 fs with minimum satellite production.

Soluble complement receptor type 1 inhibits complement system activation and improves motor function in acute spinal cord injury.

STUDY DESIGN: Spinal cord injured rat model, treated with soluble complement receptor type 1 (sCR1). SETTING: Experimental Animal Department of China Medical University, Shenyang, China. OBJECTIVES: Soluble CR1 is a powerful inhibitor of complement activation. In this study, we investigate the effectiveness of sCR1 on spinal cord injury (SCI) in rats. METHODS: Spinal cord injury was induced in Sprague-Dawley rats. Three experimental groups were examined; the sCR1 group was administered sCR1 at 1 h after the SCI, whereas the control group was administered saline at 1 h after SCI and the sham group underwent a sham operation without SCI or administration. The expressions of C9 and CD59 in the injured spinal cords were evaluated by immunohistochemistry, and numbers of positive cells counted. Furthermore, myeloperoxidase (MPO) activity and motor function were evaluated in each group. RESULTS: At all postoperative time points, the numbers of C9- and CD59-positive cells in the sCR1 group were reduced compared with the control group and MPO activity was significantly decreased compared with both other groups. Moreover, the Basso, Beattie and Bresnahan score for the sCR1 group was significantly improved as compared with that of the control group after 7 days postoperatively. CONCLUSION: Soluble CR1 decreases inflammation reactions by inhibiting activation of the complement system and improves motor function after acute SCI.

Digital imaging in transmission electron microscopy.

The digital revolution currently under way, as evidenced by the rapid development of the Internet and the world-wide-web technologies, is undoubtedly impacting the field of transmission electron microscopy (TEM). Digital imaging systems based on charge-coupled device (CCD) technologies, with pixel array size up to 2 k x 2 k at the present and increasing, are available for TEM applications and offer many attractions. Is it time to phase out film cameras on TEMs and close the darkrooms for good? This paper reviews digital imaging technologies for TEM at different voltages, and contrasts the performance of digital imaging systems with that of TEM film. The performance characteristics of CCD-based digital imaging systems, as well as methods for assessing them, are discussed. Other approaches to digital imaging are also briefly reviewed.

Multiport-readout frame-transfer 5 megapixel CCD imaging system for TEM applications.

A multiport-readout, frame-transfer charge-coupled device (CCD) digital imaging system has been successfully developed and tested for intermediate-high-voltage electron microscopy (IVEM) applications up to 400 keV. The system employs a back-thinned CCD with 2560 x 1960 pixels and a pixel size of 24 microm x 24 microm. In the current implementation, four of the eight on-chip readout ports are used in parallel each operating at a pixel rate of 1- or 2-MHz so that the entire CCD array can be read out in as short as 0.6 s. The frame-transfer readout functions as an electronic shutter which permits the rapid transfer of charges in the active pixels to four masked buffers where the charges are readout and digitized while the active area of the CCD is integrating the next frame. With a thin film-based phosphor screen and a high-performance lens relay, the system has a conversion factor of 2.1 digital units per incident electron at 400 keV, and a modulation transfer function value of 14% at the Nyquist frequency.

Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons.

A video-rate (30 frames/s) scanning two-photon excitation microscope has been successfully tested. The microscope, based on a Nikon RCM 8000, incorporates a femtosecond pulsed laser with wavelength tunable from 690 to 1050 nm, prechirper optics for laser pulse-width compression, resonant galvanometer for video-rate point scanning, and a pair of nonconfocal detectors for fast emission ratioing. An increase in fluorescent emission of 1.75-fold is consistently obtained with the use of the prechirper optics. The nonconfocal detectors provide another 2.25-fold increase in detection efficiency. Ratio imaging and optical sectioning can therefore be performed more efficiently without confocal optics. Faster frame rates, at 60, 120, and 240 frames/s, can be achieved with proportionally reduced scan lines per frame. Useful two-photon images can be acquired at video rate with a laser power as low as 2.7 mW at specimen with the genetically modified green fluorescent proteins. Preliminary results obtained using this system confirm that the yellow "cameleons" exhibit similar optical properties as under one-photon excitation conditions. Dynamic two-photon images of cardiac myocytes and ratio images of yellow cameleon-2.1, -3.1, and -3.1nu are also presented.

ASIC-based event-driven 2D digital electron counter for TEM imaging.

A two-dimensional application specific integrated circuit (ASIC) based detector, designed for X-ray protein crystallography, has been tested to determine its suitability as a direct electron detector for TEM imaging in the voltage range of 20-400 keV. Several markedly different properties of this device distinguish it from the charge coupled device (CCD) detectors: (1) the ASIC detector can be used directly under electron bombardment in the voltage range stated above, therefore requiring no scintillator screen; (2) each active pixel of the device is an electron counter and generates digital output independently; (3) the readout of the device is frameless and event driven; (4) the device can be operated at the room temperature and is nearly noise free; and (5) the counting dynamic range of the device is virtually unlimited. It appears that an imaging system based on this type of device would be ideal for low-dose TEM imaging and online diffraction observation and recording, as well as more conventional imaging, providing the many advantages of direct digital readout for almost all applications.

Optimization of thin-foil based phosphor screens for CCD imaging in TEM in the voltage range of 80-400 kV.

The voltage dependence characteristics of thin-foil based phosphor screens in the thickness range of approximately 10-60 microns are examined for CCD imaging in transmission electron microscopy (TEM) in the voltage range of 80-400 kV. The brightest screen is obtained with a P20 layer of about 12 microns at 80 kV, and a thicker screen lowers both the screen brightness and resolution. The thickness of the brightest screen is higher at higher voltage, but other considerations for a practical CCD imaging system suggest that the P20 layer should not be greater than approximately 18 microns for the voltage range stated above.

In vivo calcineurin crystals formed using the baculovirus expression system.

Calcineurin is a heterodimeric phosphatase involved in the signal transduction of antigen-activated T cells. Coexpression of its two subunits, the regulatory subunit from human and the catalytic subunit from Neurospora crassa in cultured insect cells using the baculovirus expression system results in the formation of very large crystals in the cytoplasm. The crystals are formed initially in vesicles, but their subsequent growth appears to be uninhibited and continues without the need of an enclosing membrane until the host cell lyses. Although these in vivo crystals are low in population, ranging only 0-3 per cell, they are extremely large, over 10 mu m in some cases. Biochemical assays confirm their calcineurin origin, with the regulatory subunit incorporated being myristoylated, although both the myristoylated and unmyristoylated forms are expressed. The lattice structure of the in vivo crystals, with a spacing of 5.5 nm, is preserved with the regular electron microscopic (EM) specimen preparation procedure.

Conditions for electron tomographic data acquisition.

The conventional algorithms employed in electron microscope tomography require that the series of images obtained from different orientations of the specimen each represent a parallel orthographic projection under uniform magnification. Electron microscope optics can produce distortions in images that may affect the accuracy of a tomographic reconstruction. These distortions result in images with differential rotation and magnification of regions of a thick or highly tilted specimen located at different distances with respect to the plane of focus. The distortions increase in magnitude and may significantly affect the accuracy of the tomographic data when images are acquired under current center misalignment or non-eucentric positioning of the specimen. An additional source of error can be introduced if the condenser lenses are adjusted to compensate for the intensity attenuation due to the increased beam path length as the specimen is tilted. The change in beam crossover position due to adjustment of the condensers alters the beam divergence and may introduce systematic changes in magnification over the tilt series. The process of alignment of the tilt series will propagate these errors to more central regions of the image. In most cases, with proper consideration, it is possible to minimize these errors to levels where they will have negligible effects on the resolution of the tomographic reconstruction.

Stereoscopy by tilted illumination in transmission electron microscopy.

We describe a new technique for stereoscopic observation in transmission electron microscopy, employing tilted illumination. A triple-hole objective aperture is used so that the bright beam can pass through with or without tilt. Stereo views can be acquired by tilting the illumination such that the bright beam passes through a pair of symmetrically arranged apertures alternately. The advantages of this technique as compared to the commonly used method of single-axis tilt are: (i) greater speed, potentially at or close to video rate so that live 3D observation is possible; (ii) elimination of specimen movement associated with stage tilt; and (iii) perspective views corresponding to multiple tilting axes which can be realized by installing more aperture-holes and choosing their positions properly. The main limitation is that the angle of tilt is limited by the extent to which the astigmatism, introduced by beam tilt, can be compensated by the objective lens stigmators.

Performance of thin foil scintillating screen for transmission electron microscopy.

Scintillating screens made by coating a thin metal foil with a layer of phosphor appear to be attractive for transmission electron microscopy applications. We report here the brightness and resolution in the voltage range of 100-400 kV of such a screen made of a 10 microns layer of P20 phosphor on a 2 microns Al foil. Both brightness and resolution are superior to that of a screen made by coating a glass plate with a similar layer of phosphor. An exciting property of such a device is that its resolution improves slightly at higher voltages. This, combined with its excellent resistivity to radiation damage and stability under the electron beam, makes it a good candidate for high-voltage applications.

Performance characteristics of radioluminescent fiber optics as electron scintillators.

Terbium-activated radioluminescent fiber-optic face plates have been studied for their suitability as electron scintillators. The energy conversion efficiency of such material is determined to be about 2.5% in the electron energy range of 100-400 keV tested. The resolution is approximately 40 microns at electron energy of 100 keV, as measured by the rise width of an edge. The main advantage of such a device is the potential simplification of CCD camera designs, since it functions both as a scintillator as well as a light guide so that it can be coupled direct to a CCD detector.

Telemicroscopy.

The feasibility of operating an electron microscope from a remote location using a high-speed computer network has been investigated. A 400 kV electron microscope was modified so that most microscope parameters and stage movement (x, y, z and tilt), can be controlled by a local host computer. The host computer also controls acquisition of digital images from the microscope from either a TV or a slow-scan CCD camera. A computer located at a remote site may send commands to the host computer via a fiber optic network to receive digital images and information concerning the status of microscope. Such a system has been demonstrated at the SIGGRAPH 1992 conference in Chicago and the Supercomputing 92 conference in Minneapolis, where we remotely collected three-dimensional image data sets using the electron microscope in San Diego. This approach, referred to as "telemicroscopy", promises to make possible live microscopy teleconferencing and thereby increase the accessibility and effective usage of the limited number of specialized microscopy resources.

High-sensitivity lens-coupled slow-scan CCD camera for transmission electron microscopy.

A lens-coupled slow-scan CCD camera has been built for transmission electron microscopy (TEM) applications. In this design, a leaded glass window, which is coated with a 20 microns layer of red P20 phosphor, is mounted on the bottom of the microscope. A lens assembly and mirror prism, located outside the microscope vacuum below the leaded glass, relays the image onto a back-thinned 1kx1k charge-coupled device (CCD) detector. This CCD is electronically cooled to below -30 degrees C during operation. It is found that X-ray irradiation, generally found to be annoying in fiber-optically coupled CCD cameras, is completely eliminated by this configuration. The collection efficiency of this system, although not as high as some of the fiber-optically coupled CCD cameras, is high enough to achieve single-electron sensitivity under a high-gain mode.

[Reversion of eye position after surgical correction of exotropia].

The authors analysed the reversion of eye position in 62 postoperative patients of concomitant exotropia. The results indicated the presence of a correlation with the preoperative condition of binocular vision, the type of strabismus, and the age of patient. The authors opened that the amplitude of surgical correction should depend on these factors, and prudence should be exercised in recommending surgical intervention for mild intermittent exotropia in children.

[Tubal ligation via laparoscope: report of 84 cases (author's transl)].

PIP: 84 cases of tubal ligation with satisfactory results were performed using the instruments and procedures recently developed. The instrument was patterned after a rectal speculum and consisted of a laparoscope and a tube-holding forceps. The procedure began by lowering the laparoscope into the peritoneal cavity through a 2 cm vertical incision and moving the laparoscope gently to locate the oviduct along the anterior wall of the uterus. The fimbriae of the oviduct were brought into view, and the laparoscope was accurately focused on the juction of the isthmus of the oviduct and the uterine fundus. The tube-holding forceps were then inserted to pick up and ligate the oviduct at the junction. If the uterus was small or occupied a posterior position, the oviduct was located by the laparoscope via the broad ligament rather than the anterior wall of the uterus because the laparoscope could be easily blocked by intestines and the greater omentum. This method was found to be accurate, reliable, and to greatly reduce any complications. It was applicable regardless of the size or position of the uterus. The operation was easily and rapidly performed, required simple instruments, and could be widely used in rural areas.^ieng