Effect of the surrounding environment on electron beam irradiation damage of enhanced green fluorescent protein.

Fluorescent proteins exhibit fluorescence and photoconversion, which are used to study biological phenomena. Among these, enhanced green fluorescent protein (EGFP) emits cathodoluminescence when irradiated with electron beams; this phenomenon has numerous applications in new research tools for biological phenomena. However, bleaching during electron irradiation is a major problem.

Application of Hilbert-differential phase contrast to scanning transmission electron microscopy.

We report a novel class of scanning transmission electron microscopy with Hilbert-differential phase contrast (HDP-STEM) that displays nanostructures of thin samples in a topographical manner. A semicircular π-phase plate (PP) was used as an optical device for manipulating electron waves in HDP-STEM. This is the different design from the Zernike PP used in our previous phase plate STEM (P-STEM), but both must be placed in the front focal plane of the condenser lens.

First step toward complex observations by 4D-STEM with phase plate.

Quantitative measurements by electron microscopy are becoming increasingly important because we are often concerned with establishing quantitative relationships between the properties and structures of materials. This paper presents a method to derive the scattering and phase contrast components from scanning transmission electron microscope (STEM) images using a phase plate and two-dimensional electron detector and to quantitatively evaluate the amount of phase modulation. The phase-contrast transfer function (PCTF) modifies the phase contrast because it is not unity over all spatial frequency regions; therefore, the amount of phase modulation observed in the image becomes smaller than the actual value.

Toward complex observation in electron microscopy using two-dimensional electron detector coupled with phase plate STEM.

A two-dimensional (2D) detector was used to construct phase plate STEM (P-STEM) images. Phase-contrast can be enhanced by the electron intensity inside the hole region of a thin film phase plate. The electron intensity outside the hole region also provides a dark image contrast, which is inconsistent with the weak phase object approximation.

Evaluation of electron radiation damage to green fluorescent protein.

Green fluorescent protein (GFP) emits light when irradiated by not only light but also electrons. This electron-induced light emission called cathodoluminescence (CL) can be used to realize a high-resolution light emission microscopy based on the irradiation of a very narrow electron beam. To implement CL mapping in life sciences the investigation of the damage resistance of GFP to electron irradiation needs to be clarified.

Cathodoluminescence of green fluorescent protein exhibits the redshifted spectrum and the robustness.

Green fluorescent protein (GFP) and its variants are an essential tool for visualizing functional units in biomaterials. This is achieved by the fascinating optical properties of them. Here, we report novel optical properties of enhanced GFP (EGFP), which is one of widely used engineered variants of the wild-type GFP.

Investigation of hydrated smectite microstructure through wet environmental transmission electron microscopy.

Water is an essential constituent of all biological materials as well as many non-biological materials. Not only the removal of water may result in undesirable morphological and structure change, the inability to sustain the hydrated conditions in the microscope also prevents the study of reactions which take place in aqueous environment. In order to overcome these problems we used wet environmental-cell transmission electron microscopy TEM (WETEM).

In planta functions of cytochrome P450 monooxygenase genes in the phytocassane biosynthetic gene cluster on rice chromosome 2.

In response to environmental stressors such as blast fungal infections, rice produces phytoalexins, an antimicrobial diterpenoid compound. Together with momilactones, phytocassanes are among the major diterpenoid phytoalexins. The biosynthetic genes of diterpenoid phytoalexin are organized on the chromosome in functional gene clusters, comprising diterpene cyclase, dehydrogenase, and cytochrome P450 monooxygenase genes.

Contrast enhancement of nanomaterials using phase plate STEM.

Visualizing materials composed of light elements is difficult, and the development of an imaging method that enhances the phase contrast of such materials has been of much interest. In this study, we demonstrate phase-plate scanning transmission electron microscopy (P-STEM), which we developed recently, and its application to nanomaterials. An amorphous carbon film with a small hole in its center was used to control the phase of incident electron waves, and the phase-contrast transfer function (PCTF) was modified from sine-type to cosine-type.

Cathodoluminescence and Electron-Induced Fluorescence Enhancement of Enhanced Green Fluorescent Protein.

Becaues the spatial resolution of fluorescence microscopy is not high enough to study the molecular level of relationship between the structure and function of biological specimens, correlative light and electron microscopy has been used for this purpose. Another possibility for a high-resolution light microscopy is cathodoluminescence microscopy. Here, we report a new phenomenon, the electron-induced activation of luminescence (cathodoluminescence) and electron-enhanced fluorescence for the enhanced green fluorescent protein (EGFP).

Electron microscopic recording of myosin head power stroke in hydrated myosin filaments.

Muscle contraction results from cyclic attachment and detachment between myosin heads and actin filaments, coupled with ATP hydrolysis. Despite extensive studies, however, the amplitude of myosin head power stroke still remains to be a mystery. Using the gas environmental chamber, we have succeeded in recording the power stroke of position-marked myosin heads in hydrated mixture of actin and myosin filaments in a nearly isometric condition, in which myosin heads do not produce gross myofilament sliding, but only stretch adjacent elastic structures.

Phase-contrast scanning transmission electron microscopy.

This report introduces the first results obtained using phase-contrast scanning transmission electron microscopy (P-STEM). A carbon-film phase plate (PP) with a small center hole is placed in the condenser aperture plane so that a phase shift is introduced in the incident electron waves except those passing through the center hole. A cosine-type phase-contrast transfer function emerges when the phase-shifted scattered waves interfere with the non-phase-shifted unscattered waves, which passed through the center hole before incidence onto the specimen.

Pattern visual evoked potentials elicited by organic electroluminescence screen.

Purpose: To determine whether organic electroluminescence (OLED) screens can be used as visual stimulators to elicit pattern-reversal visual evoked potentials (p-VEPs).

Method: Checkerboard patterns were generated on a conventional cathode-ray tube (S710, Compaq Computer Co., USA) screen and on an OLED (17 inches, 320 × 230 mm, PVM-1741, Sony, Tokyo, Japan) screen.

Binocular interaction of visually evoked cortical potentials elicited by dichoptic binocular stimulation.

To analyze the interaction of cortical potentials elicited by dichoptic stimulation of the dominant and fellow eyes at different frequencies, a pair of programmed power supply units were used to drive a light emitting diode (LED) mounted in the right and left eyes of light-proof goggles to elicit the visually evoked cortical responses (VECPs). The right eye was stimulated at 11.5 Hz and the left eye at 11.

Comparisons of pattern visually evoked potentials elicited by different response time liquid crystal display screens.

Purpose: To evaluate the usefulness of a liquid crystal display (LCD) with higher driving frequency and shorter response time (2 ms) as a visual stimulator to elicit pattern reversal visually evoked potentials (p-VEPs).

Method: p-VEPs were recorded from 12 eyes of 12 healthy volunteers (28.3 ± 9 years).

Liquid crystal display screens as stimulators for visually evoked potentials: flash effect due to delay in luminance changes.

Purpose: The cathode-ray tube (CRT) screen has recently been replaced by liquid crystal display (LCD) screens as visual stimulators for pattern-reversal visually evoked potentials (p-VEPs). The aim of the study was to evaluate the usefulness of LCD screen to elicit p-VEPs.

Methods: The waveforms of the p-VEPs elicited by a LCD panel were compared with those elicited by a conventional CRT screen.

A carbon sandwich environmental cell for wet specimens.

A carbon sandwich environmental cell for wet specimens was developed for environmental transmission electron microscopy (E-TEM). A plastic film with many holes was used to form carbon capsules. The carbon sandwich environmental cells were composed of the spacer and two carbon films and enclosing the samples and experimental solution.

Direct observation of biological molecules in liquid by environmental phase-plate transmission electron microscopy.

We have been developing a combination method for environmental TEM (E-TEM) and phase-plate TEM (P-TEM) that enables direct observations of the structure of biological molecules in aqueous solution. It is clearly demonstrated that the biological molecules in a water layer can be imaged by the combined method without any stain. The spatial resolution obtained in the present study was about 10nm.

Contrast enhancement in the phase plate transmission electron microscopy using an objective lens with a long focal length.

A new optical condition using an objective lens (OL) of a long focal length (objective mini lens: OM) was tested to enhance image contrast in phase plate transmission electron microscopy (P-TEM). A phase plate was set on the selected area aperture plane where diffraction patterns were formed under the optical condition using the OM. A phase shift by the phase plate was added to the electron waves to visualize phase objects.

Electron microscopic evidence for the myosin head lever arm mechanism in hydrated myosin filaments using the gas environmental chamber.

Muscle contraction results from an attachment-detachment cycle between the myosin heads extending from myosin filaments and the sites on actin filaments. The myosin head first attaches to actin together with the products of ATP hydrolysis, performs a power stroke associated with release of hydrolysis products, and detaches from actin upon binding with new ATP. The detached myosin head then hydrolyses ATP, and performs a recovery stroke to restore its initial position.

[Burden sharing with co-medical personnel in the surgical area: present and future situations. The clinical engineer's position].

It has been 22 years since clinical engineers first made their appearance as experts in medical equipment used to save patients' lives. At that time, legal revisions, ministerial ordinances, and regulations were established, with the final step awaiting a change in the medical environment. The pressing problem of a serious shortage of physicians is now apparent.

Development of a surface conductivity measurement system for ultrahigh vacuum transmission electron microscope.

The surface conductivity measurement system using a micro-four-point probe (M4PP) had been developed for the ultrahigh vacuum transmission electron microscope (UHV-TEM). Since the current distribution in the sample crystals during the current voltage measurement by the M4PP is localized within the depth of several micrometers from the surface, the system is sensitive to the surface conductivity, which is related with the surface superstructure. It was installed in the main chamber of the TEM and the surface conductivity can be measured in situ.

[STD in the eye].

In this paper, we review sexually transmitted diseases (STD) involving the eye. Recently conjunctivitis due to Chlamydia trachomatis in children and adults is increasing, and that of Neisseria gonorrhoeae resistant to multiple antibiotics has attracted special attention in our country. Syphilis has many ocular manifestations such as keratitis, iridocyclitis, retinochorioiditis, and neuritis, etc.

Direct demonstration of the cross-bridge recovery stroke in muscle thick filaments in aqueous solution by using the hydration chamber.

Despite >50 years of research work since the discovery of sliding filament mechanism in muscle contraction, structural details of the coupling of cyclic cross-bridge movement to ATP hydrolysis are not yet fully understood. An example would be whether lever arm tilting on the myosin filament backbone will occur in the absence of actin. The most direct way to elucidate such movement is to record ATP-induced cross-bridge movement in hydrated thick filaments.

Clinical evaluation of blue-light blocker foldable acrylic IOL (HMY).

Efficacy and safety of blue-light-filtering acrylic IOL (HMY) were assessed and proven from 3 months to 1 year of postoperative follow-up in a clinical study performed on 80 eyes of 45 patients. Postoperative corrected visual acuity of all the eyes was 6/12 or better and 78.8% of them achieved higher than 6/6.