The immune system maintains appropriate cell numbers through regulation of cell proliferation and death. Normal tissue distribution of lymphocytes is maintained through expression of specific adhesion molecules and chemokine receptors such as L-selectin and CCR7, respectively. Lymphocyte insufficiency or lymphopenia induces homeostatic proliferation of existing lymphocytes to increase cell numbers. Interestingly, homeostatic proliferation of T lymphocytes induces a phenotypic change from naïve- to memory-type cell. Naïve T cells recirculate between blood and lymphoid tissues whereas memory T cells migrate to nonlymphoid sites such as skin and gut. To assess effects of homeostatic proliferation on migratory ability of T cells, a murine model of lymphopenia-induced homeostatic proliferation was used. Carboxyfluorescein diacetate, succinimidyl ester-labeled wild-type splenocytes were adoptively transferred into recombination activation gene-1-deficient mice and analyzed by flow cytometry, in vitro chemotactic and in vivo migration assays, and immunofluorescence microscopy. Homeostatically proliferated T cells acquired a mixed memory-type CD44high L-selectinhigh CCR7low phenotype. Consistent with this, chemotaxis to secondary lymphoid tissue chemokine in vitro was reduced by 22%–34%. By contrast, no differences were found for migration or entry into lymph nodes during in vivo migration assays. Therefore, T lymphocytes that have undergone homeostatic proliferation recirculate using mechanisms similar to naïve T cells.

Sertoli cells are very important to spermatogenesis homeostasis because they control germ cell proliferation, differentiation, and death. Damages to Sertoli cells cause germ cell death and affect fertility. Etoposide is a potent chemotherapeutic drug largely used against a variety of cancers. However, this drug also kills normal cells, especially those undergoing rapid proliferation. In the testis, etoposide acts predominantly on intermediate and type B spermatogonia. Etoposide was shown to permanently alter Sertoli cell function when administered to prepubertal rats. Based on this, we decided to investigate whether etoposide can affect Sertoli cell morphology. For this, 25-day-old rats were treated with etoposide during 8 consecutive days and killed at 32, 45, 64, 127, and 180 days old. Testes were fixed in Bouin's liquid or in a mixture of 2.5% glutaraldehyde and 2% formaldehyde for analysis under light and electron microscopes, respectively. Sertoli cells showed morphological alterations such as the presence of chromatin clumps close to the nuclear membrane, nucleus displacement, and cytoplasmic vacuolization. Some Sertoli cells also showed nuclear and cytoplasmic degenerative characteristics, suggesting that etoposide causes severe damages to Sertoli cell.

Disposable coordinate standard (CS) chips were fabricated by the ejection of melted polystyrene into a metal mold. The CS chip surface was divided into four parts different in height and width. The edge lines of these parts could be recognized as straight lines 2 μm in width in the microscope view and used as the X and Y axes for the culture dish. The CS chip was attached on the bottom of a culture dish outside. Then the dish was set on the microscope stage and moved by means of a motorized automatic stage. The X-Y coordinates of many single-cells in a culture dish were registered, respectively. Once registered, any single-cell could instantly be brought to the center of the microscope view even after displacing the dish from the stage for a while and setting it again on the stage. Therefore, experimenters can easily search any single-cell in any culture dish on any microscope at any time. Such a system is remarkably useful for various modes of single-cell experiment and named “Suguwaculture,” which means “instantly” (“sugu” in Japanese) + “recognizable” (“wakaru” in Japanese) + “culture” (during culture).

Light-emitting diodes (LEDs) can be easily and inexpensively integrated into modern light microscopes. There are numerous advantages of LEDs as illumination sources; most notably, they provide brightness and spectral control. We demonstrate that for transmitted light imaging, an LED can replace the traditional tungsten filament bulb while offering longer life; no color temperature change with intensity change; reduced emission in the infrared region, which is important for live cell imaging; and reduced cost of ownership. We show a direct substitution of the typical tungsten bulb with a commercially available LED and demonstrated the color stability by imaging a histology section over a wide range of light intensities. For fluorescent imaging, where the typical illumination sources are mercury or xenon lamps, we demonstrate that LEDs offer advantages of providing a longer lifespan, having a more constant intensity output over time, more homogeneous illumination, and significantly lower photon dose. Our LED equipped system was used to image and deconvolve dual fluorescently labeled cells, as well as image cells undergoing mitosis expressing green fluorescent protein–histone 2B complex. The timing of the stages of mitosis is well established as an indicator of cell viability.

A significant portion of Mesozoic amber is fully opaque. Biological inclusions in such amber are invisible even after polishing, leading to potential bias in paleoecological and phylogenetic studies. Until now, studies using conventional X-ray microtomography focused on translucent or semi-opaque amber. In these cases, organisms of interest were visualized prior to X-ray analyses. It was recently demonstrated that propagation phase contrast X-ray synchrotron imaging techniques are powerful tools to access invisible inclusions in fully opaque amber. Here we describe an optimized synchrotron microradiographic protocol that allowed us to investigate efficiently and rapidly large amounts of opaque amber pieces from Charentes (southwestern France). Amber pieces were imaged with microradiography after immersion in water, which optimizes the visibility of inclusions. Determination is not accurate enough to allow precise phylogenetic studies, but provides preliminary data on biodiversity and ecotypes distribution; phase contrast microtomography remains necessary for precise determination. Because the organisms are generally much smaller than the amber pieces, we optimized local microtomography by using a continuous acquisition mode (sample moving during projection integration). As tomographic investigation of all inclusions is not practical, we suggest the use of a synchrotron for a microradiographic survey of opaque amber, coupled with microtomographic investigations of the most valuable organisms.

Cold gas dynamic spray (cold spray) is a rapid deposition technology in which particles deposit at velocities above the speed of sound (∼340 ms−1). Generally, porosity forms in cold spray deposits due to insufficient deformation of particles. In this study, the unique capability of the X-ray microscopy and microtomography is utilized to visualize the internal structure of deposited material. The results show that this characterization technique successfully reveals porosities in the cold spray commercial purity (CP) titanium structure. Furthermore, microtomography images confirmed the experimental results for porosity measurements in which helium (compared with nitrogen) as carrier gas significantly decreases porosity in cold spray CP titanium.

Growth of bismuth oxide (most probably Bi2O3) was observed in situ in a transmission electron microscope. Bi liquid particles were dispersed on the substrates of diamond or SiO2. Introduction of oxygen up to ∼5 × 10−4 Pa resulted in formation of bismuth oxide (most probably Bi2O3) whiskers. The growth mechanism of the whisker was discussed in terms of a vapor-liquid-solid (VLS) mechanism. It is suggested that the liquid droplet of Bi acts as a physical catalyst for growth of bismuth oxide (most probably Bi2O3) whiskers.

Electromagnetic response of individual boron nitride nanotubes (BNNTs) has been studied by spatially resolved electron energy loss spectroscopy (EELS). We demonstrate how dedicated EELS methods using subnanometer electron probes permit the analysis of local dielectric properties of a material on a nanometer scale. The continuum dielectric model has been used to analyze the low-loss EEL spectra recorded from these tubes. Using this model, we demonstrate the weak influence of the out-of-plane contribution to the dielectric response of BNNTs. The optical gap, which can be deduced from the measurements, is found to be equal to 5.8 ± 0.2 eV, which is close to that of the hexagonal boron nitride. This value is found to be independent of the nanotubes configuration (diameter, helicity, number of walls, and interaction between the different walls).