Microscopy最新文献

{"title":"Histochemical assessment on the cellular interplay of vascular endothelial cells and septoclasts during endochondral ossification in mice","authors":"Erika Tsuchiya;Tomoka Hasegawa;Hiromi Hongo;Tomomaya Yamamoto;Miki Abe;Taiji Yoshida;Shen Zhao;Kanako Tsuboi;Nobuyuki Udagawa;Paulo Henrique Luiz de Freitas;Minqi Li;Yoshimasa Kitagawa;Norio Amizuka","doi":"10.1093/jmicro/dfaa047","DOIUrl":"10.1093/jmicro/dfaa047","url":null,"abstract":"This study was aimed to verify the cellular interplay between vascular endothelial cells and surrounding cells in the chondro-osseous junction of murine tibiae. Many CD31-positive endothelial cells accompanied with Dolichos Biflorus Agglutinin lectin-positive septoclasts invaded into the hypertrophic zone of the tibial epiphyseal cartilage. MMP9 immunoreactive cytoplasmic processes of vascular endothelial cells extended into the transverse partitions of cartilage columns. In contrast, septoclasts included several large lysosomes which indicate the incorporation of extracellular matrices despite no immunopositivity for F4/80—a hallmark of macrophage/monocyte lineage. In addition, septoclasts were observed in c-fos\u0000^-/-\u0000 mice but not in Rankl\u0000^-/-\u0000 mice. Unlike c-fos\u0000^-/-\u0000 mice, Rankl\u0000^-/-\u0000 mice showed markedly expanded hypertrophic zone and the irregular shape of the chondro-osseous junction. Immunoreactivity of platelet-derived growth factor-bb, which involved in angiogenic roles in the bone, was detected in not only osteoclasts but also septoclasts at the chondro-osseous junction. Therefore, septoclasts appear to assist the synchronous vascular invasion of endothelial cells at the chondro-osseous junction. Vascular endothelial cells adjacent to the chondro-osseous junction possess endomucin but not EphB4, whereas those slightly distant from the chondro-osseous junction were intensely positive for both endomucin and EphB4, while being accompanied with ephrinB2-positive osteoblasts. Taken together, it is likely that vascular endothelial cells adjacent to the chondro-osseous junction would interplay with septoclasts for synchronous invasion into the epiphyseal cartilage, while those slightly distant from the chondro-osseous junction would cooperate with osteoblastic activities presumably by mediating EphB4/ephrinB2. Mini-abstract Our original article demonstrated that vascular endothelial cells adjacent to the chondro-osseous junction would interplay with septoclasts for synchronous invasion into the epiphyseal cartilage, while those slightly distant from the chondro-osseous junction would cooperate with osteoblastic activities presumably by mediating EphB4/ephrinB2. (A figure that best represents your paper is Fig. 5c)","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa047","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38281522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Three-dimensional culture of a pancreatic cancer cell line, SUIT-58, with air exposure can reflect the intrinsic features of the original tumor through electron microscopy","authors":"Nobuyasu Takahashi;Fumiyo Aoyama;Akira Sawaguchi","doi":"10.1093/jmicro/dfaa046","DOIUrl":"10.1093/jmicro/dfaa046","url":null,"abstract":"Mini-abstract: Application of a three-dimensional culture system with air exposure facilitates the formation of large cell spheres possessing cribriform glands and producing mucin in the collagen gel. Transmission electron microscopy revealed the formation of microvilli and junctional complexes at the apical side of the cell. This study aimed to reproduce the characteristics of original adenocarcinoma tumors in vitro. The pancreatic cell line, SUIT-58, derived from a moderately differentiated adenocarcinoma of metastatic pancreatic cancer was used. The cells have a sheet structure in conventional cell culture without forming glands or exhibiting mucin production in the lumen. First, the necessity of scaffolds to create an adenocarcinoma-like microenvironment for SUIT-58 pancreatic cancer cells was assessed. Compared with conventional culture plates, the use of type I collagen as a scaffold played an important role in the formation of densely congested microvilli, as observed through scanning electron microscopy. As gland formation is one of the features of adenocarcinoma, we also assessed gland formation. Use of a recently developed three-dimensional culture system with air exposure resulted in the formation of large cell spheres possessing cribriform glands, which released mucin into the lumen. Transmission electron microscopy also revealed the formation of microvilli in the lumen of the glands and junctional complex at the intercellular part, which were similar to those observed in xenografts. These findings indicate that an in vitro three-dimensional culture system with air exposure reflects the intrinsic features of the original tumor, suggesting that this culture system could be useful for preliminary research of certain cancers.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38261037","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phase detection limits in off-axis electron holography from pixelated detectors: gain variations, geometric distortion and failure of reference-hologram correction","authors":"Martin Hÿtch;Christophe Gatel","doi":"10.1093/jmicro/dfaa044","DOIUrl":"10.1093/jmicro/dfaa044","url":null,"abstract":"We investigate the effect that recording off-axis electron holograms on pixelated detectors, such as charge-coupled devices (CCD) and direct-detection devices (DDD), can have on measured amplitudes and phases. Theory will be developed for the case of perfectly uniform interference fringes illuminating an imperfect detector with gain variations and pixel displacements. We will show that both these types of defect produce a systematic noise in the phase images that depends on the position of the holographic fringes with respect to the detector. Subtracting a reference hologram from the object hologram will therefore not remove the phase noise if the initial phases of the two holograms do not coincide exactly. Another finding is that pi-shifted holograms are much less affected by gain variations but show no improvement concerning geometric distortions. The resulting phase errors will be estimated and simulations presented that confirm the theoretical developments.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa044","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38229752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Three-dimensional structure analysis of melanocytes and keratinocytes in senile lentigo","authors":"Yuki Mizutani;Mika Yamashita;Rie Hashimoto;Toru Atsugi;Akemi Ryu;Akinobu Hayashi;Yukiko Rikimaru-Nishi;Keisuke Ohta","doi":"10.1093/jmicro/dfaa054","DOIUrl":"10.1093/jmicro/dfaa054","url":null,"abstract":"Senile lentigo or age spots are hyperpigmented macules of skin that commonly develop following long-term exposure to ultraviolet radiation. This condition is caused by accumulation of large numbers of melanosomes (melanin granules) produced by melanocytes within neighboring keratinocytes. However, there is still no consensus regarding the melanosome transfer mechanism in senile lentigo. To date, most pathohistological studies of skin have been two-dimensional and do not provide detailed data on the complex interactions of the melanocyte–keratinocyte network involved in melanosome transfer. We performed a three-dimensional reconstruction of the epidermal microstructure in senile lentigo using three different microscopic modalities to visualize the topological melanocyte–keratinocyte relationship and melanosome distribution. Confocal laser microscopy images showed that melanocyte dendritic processes are more frequently branched and elongated in senile lentigo skin than in normal skin. Serial transmission electron micrographs showed that dendritic processes extend into intercellular spaces between keratinocytes. Focused ion beam-scanning electron micrographs showed that dendritic processes in senile lentigo encircle adjacent keratinocytes and accumulate large numbers of melanosomes. Moreover, melanosomes transferred to keratinocytes are present not only in the supranuclear area but throughout the perinuclear area except on the basal side. The use of these different microscopic methods helped to elucidate the three-dimensional morphology and topology of melanocytes and keratinocytes in senile lentigo. We show that the localization of melanosomes in dendritic processes to the region encircling recipient keratinocytes contributes to efficient melanosome transfer in senile lentigo.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa054","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38432747","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Wave field reconstruction and phase imaging by electron diffractive imaging","authors":"Jun Yamasaki","doi":"10.1093/jmicro/dfaa063","DOIUrl":"10.1093/jmicro/dfaa063","url":null,"abstract":"In electron diffractive imaging, the phase image of a sample is reconstructed from its diffraction intensity through iterative calculations. The principle of this method is based on the Fourier transform relation between the real-space wave field transmitted by the sample and its Fraunhofer diffraction wave field. Since Gerchberg’s experimental work in 1972, various advancements have been achieved, which have substantially improved the quality of the reconstructed phase images and extended the applicable range of the method. In this review article, the principle of diffractive imaging, various experimental processes using electron beams and application to specific samples are explained in detail.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa063","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38633105","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Interference and interferometry in electron holography","authors":"Ken Harada","doi":"10.1093/jmicro/dfaa033","DOIUrl":"10.1093/jmicro/dfaa033","url":null,"abstract":"This paper reviews the basics of electron holography as an introduction of the holography part of this special issue in Microscopy. We discuss the general principle of holography and interferometry regarding measurements and analyses of phase distributions, first using the optical holography. Next, we discuss physical phenomena peculiar to electron waves that cannot be realized by light waves and principles of electromagnetic field detection and observation methods. Furthermore, we discuss the interference optical systems of the electron waves and their features, and methods of reconstruction of the phase information from electron holograms, which are essential for realization of practical electron holography. We note that following this review application of electron holography will be discussed in detail in the papers of this special issue.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa033","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38088195","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of accuracy in the determination of crystal structure factors using large-angle convergent-beam electron diffraction patterns","authors":"Daisuke Morikawa;Kenji Tsuda","doi":"10.1093/jmicro/dfaa041","DOIUrl":"10.1093/jmicro/dfaa041","url":null,"abstract":"The accuracy of electron density distribution analysis using large-angle convergent-beam electron diffraction (LACBED) patterns is evaluated for different convergence angles. An orbital ordered state of FeCr\u0000<inf>2</inf>\u0000O\u0000<inf>4</inf>\u0000 is used as an example of the analysis. Ideal orbital-ordered and non-ordered states are simulated by using orbital scattering factors. LACBED patterns calculated for the orbital-ordered state were used as hypothetical experimental data sets. Electron density distribution of the Fe 3d orbitals has been successfully reconstructed with a higher accuracy from LACBED patterns with convergence angles larger than 15.2 mrad, which is 4 times as large as that for conventional convergent-beam electron diffraction patterns. Excitation of particular Bloch waves with the aid of LACBED patterns has a key role in the accurate analysis of electron density distributions.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa041","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38174171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sandwich freezing device for rapid freezing of viruses, bacteria, yeast, cultured cells and animal and human tissues in electron microscopy","authors":"Masashi Yamaguchi;Masaki Taguchi;Katsuyuki Uematsu;Azusa Takahashi-Nakaguchi;Michiyo Sato-Okamoto;Hiroji Chibana","doi":"10.1093/jmicro/dfaa049","DOIUrl":"10.1093/jmicro/dfaa049","url":null,"abstract":"We have been using sandwich freezing of living yeast and bacteria followed by freeze-substitution for observing close-to-native ultrastructure of cells. Recently, sandwich freezing of glutaraldehyde-fixed cultured cells and human tissues have been found to give excellent preservation of ultrastructure of cells and tissues. These studies, however, have been conducted using a handmade sandwich freezing device and have been limited in a few laboratories. To spread the use of this method to other laboratories, we fabricated and commercialized a new sandwich freezing device. The new device is inexpensive, portable and sterilizable. It can be used to rapid-freeze viruses, bacteria, yeast, cultured cells and animal and human tissues to a depth of 0.2 mm if tissues are prefixed with glutaraldehyde. The commercial availability of this device will expand application of rapid freezing to wide range of biological materials.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38721832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Introduction to ‘electron interference microscopy’","authors":"Ken Harada;Tadahiro Kawasaki","doi":"10.1093/jmicro/dfaa067","DOIUrl":"10.1093/jmicro/dfaa067","url":null,"abstract":"","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa067","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38626257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Holography: application to high-resolution imaging","authors":"Takeshi Kawasaki;Yoshio Takahashi;Toshiaki Tanigaki","doi":"10.1093/jmicro/dfaa050","DOIUrl":"10.1093/jmicro/dfaa050","url":null,"abstract":"Electron holography was invented for correcting aberrations of the lenses of electron microscopes. It was used to observe the atomic arrangements in crystals after decades of research. Then it was combined with a hardware aberration corrector to enable high-resolution and high-precision analysis. Its applications were further extended to magnetic observations with sub-nanometer resolution. High-resolution electron holography has become a powerful technique for observing electromagnetic distributions in functional materials.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa050","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38433926","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}