The existence of membrane contact sites (MCS) has been reported in different systems in the past decade, and their importance has been recognised by the cell biology community. Amongst all endomembrane structures, the endoplasmic reticulum (ER) plays vital roles in organising the organelle interaction network with the plasma membrane (PM), Golgi bodies, mitochondria, plastids, endosomes and autophagosomes. A number of methods have been used to study the establishment and functions of these interactions, among them, light microscopy appears to be one of the most effective approaches. Here, we present an overview of the discovery of ER-PM contact sites, and highlight the latest developments in light microscopical-based techniques that can be used for their study.

• Klíčová slova
• ER-PM contact sites, Endoplasmic reticulum, light microscopy, membrane contact sites, plants,
• MeSH
• buněčná membrána metabolismus   ultrastruktura   MeSH
• endoplazmatické retikulum metabolismus   ultrastruktura   MeSH
• fluorescenční barviva MeSH
• mikroskopie MeSH
• rostlinné buňky metabolismus   ultrastruktura   MeSH
• rostlinné proteiny metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• fluorescenční barviva MeSH
• rostlinné proteiny MeSH

Light microscopy has been used for the evaluation of the internal and external structure of dry microcapsules. The method involves surface and penetrative staining with various dyes after which the microcapsules were embedded in suitable optically translucent material. Using this method the core material, its shape and position within the microcapsules either in total or as subunits of the core are clearly distinguishable from the wall material. The surface characteristics of the microcapsules can be observed with either light or fluorescent microscopy after staining with a fluorescent dye. Furthermore, it is a relatively simple and inexpensive method by comparison with the scanning electron microscopy. The natural character of microcapsules, without any artificial structures, has been maintained. It could serve as a routine auxiliary method for complex evaluation or control of the microencapsulation process and its optimization.

• MeSH
• barvení a značení MeSH
• emulze MeSH
• mikroskopie * MeSH
• příprava léků MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• emulze MeSH

Mechanical properties of the arterial wall depend largely on orientation and density of collagen fiber bundles. Several methods have been developed for observation of collagen orientation and density; the most frequently applied collagen-specific manual approach is based on polarized light (PL). However, it is very time consuming and the results are operator dependent. We have proposed a new automated method for evaluation of collagen fiber direction from two-dimensional polarized light microscopy images (2D PLM). The algorithm has been verified against artificial images and validated against manual measurements. Finally the collagen content has been estimated. The proposed algorithm was capable of estimating orientation of some 35 k points in 15 min when applied to aortic tissue and over 500 k points in 35 min for Achilles tendon. The average angular disagreement between each operator and the algorithm was -9.3±8.6° and -3.8±8.6° in the case of aortic tissue and -1.6±6.4° and 2.6±7.8° for Achilles tendon. Estimated mean collagen content was 30.3±5.8% and 94.3±2.7% for aortic media and Achilles tendon, respectively. The proposed automated approach is operator independent and several orders faster than manual measurements and therefore has the potential to replace manual measurements of collagen orientation via PLM.

• Klíčová slova
• collagen fiber orientation, image analysis, phase correlation, polarized light microscopy,
• MeSH
• Achillova šlacha chemie   ultrastruktura   MeSH
• aorta chemie   ultrastruktura   MeSH
• kolagen analýza   ultrastruktura   MeSH
• laboratorní automatizace MeSH
• počítačové zpracování obrazu MeSH
• polarizační mikroskopie * MeSH
• prasata MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kolagen MeSH

• MeSH
• barvení a značení metody   MeSH
• buněčné jádro ultrastruktura   MeSH
• cytoplazmatická granula MeSH
• DNA fungální analýza   MeSH
• fixace tkání metody   MeSH
• glykogen analýza   MeSH
• kvasinky chemie   ultrastruktura   MeSH
• lipidy analýza   MeSH
• mikroskopie metody   MeSH
• mykologie metody   MeSH
• odběr biologického vzorku metody   MeSH
• organely ultrastruktura   MeSH
• proteiny analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• DNA fungální MeSH
• glykogen MeSH
• lipidy MeSH
• proteiny MeSH

Light-sheet fluorescence microscopy has emerged as a powerful platform for 3-D volumetric imaging in the life sciences. Here, we introduce an important step towards its use deep inside biological tissue. Our new technique, based on digital holography, enables delivery of the light-sheet through a multimode optical fibre--an optical element with extremely small footprint, yet permitting complex control of light transport processes within. We show that this approach supports some of the most advanced methods in light-sheet microscopy: by taking advantage of the cylindrical symmetry of the fibre, we facilitate the wavefront engineering methods for generation of both Bessel and structured Bessel beam plane illumination. Finally, we assess the quality of imaging on a sample of fluorescent beads fixed in agarose gel and we conclude with a proof-of-principle imaging of a biological sample, namely the regenerating operculum prongs of Spirobranchus lamarcki.

• MeSH
• design vybavení metody   MeSH
• fluorescenční mikroskopie metody   MeSH
• jehly MeSH
• optická vlákna MeSH
• optické prostředky MeSH
• osvětlení metody   MeSH
• světlo MeSH
• zobrazování trojrozměrné metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

There are various preparatory techniques for light microscopy permitting access to the inner structure of plant body and its development. Minute objects might be processed as whole-mount preparations, while voluminous ones should be separated into smaller pieces. Hereby we summarize some of the "classical" techniques to cut more voluminous objects into slices and access their inner structure either for simple anatomical analysis or for further processing (e.g., histochemistry, immunohistochemistry, in situ hybridization, enzyme histochemistry).

• MeSH
• imunohistochemie metody   MeSH
• mikroskopie * metody   MeSH
• odběr biologického vzorku metody   MeSH
• rostliny chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The tongue of an adult degu was examined by light and scanning electron microscopy. It consists of an apex, corpus, and radix and contains a lingual prominence. The aim of this study was to describe the course of muscle fascicles of the proper lingual muscle, the presence and nature of the lingual salivary glands, and particularly the appearance and distribution of the lingual papillae. Three major types of papillae have been observed: filiform, conical, and vallate. The dorsal surface of the lingual apex extends in caudally bent filiform papillae with two spines. The lingual corpus bears long filiform papillae with a single tip. The lingual radix contains crown-like papillae in the region of the prominence and conical papillae in the remaining areas. Two oval vallate papillae were discovered caudally on the lingual radix. This first description of the lingual structures in a degu could be used for comparative studies or as basic data for differentiation of lingual morphology in this species.

• Klíčová slova
• Degu, Lingual papillae, Rodents, Scanning electron microscopy, Tongue,
• MeSH
• jazyk anatomie a histologie   ultrastruktura   MeSH
• mikroskopie elektronová rastrovací * MeSH
• mikroskopie MeSH
• Octodon anatomie a histologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In higher plants, germline differentiation occurs during a relatively short period within developing flowers. Understanding of the mechanisms that govern germline differentiation lags behind other plant developmental processes. This is largely because the germline is restricted to relatively few cells buried deep within floral tissues, which makes them difficult to study. To overcome this limitation, we have developed a methodology for live imaging of the germ cell lineage within floral organs of Arabidopsis using light sheet fluorescence microscopy. We have established reporter lines, cultivation conditions, and imaging protocols for high-resolution microscopy of developing flowers continuously for up to several days. We used multiview imagining to reconstruct a three-dimensional model of a flower at subcellular resolution. We demonstrate the power of this approach by capturing male and female meiosis, asymmetric pollen division, movement of meiotic chromosomes, and unusual restitution mitosis in tapetum cells. This method will enable new avenues of research into plant sexual reproduction.

• Klíčová slova
• A. thaliana, SPIM, cell biology, flower, germline, light sheet microscopy, live cell imaging, meiosis, plant biology,
• MeSH
• Arabidopsis cytologie   růst a vývoj   MeSH
• buněčná diferenciace * MeSH
• cytogenetické vyšetření MeSH
• květy cytologie   růst a vývoj   MeSH
• mikroskopie metody   MeSH
• zárodečné buňky rostlin cytologie   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

RCH-microscopy (Relief Contrast after Hostounský) is a new method of optical microscopy in transmitted light developed with Lambda Ltd., Prague. This method was used to study bacteria, fungi including yeasts and algae at high magnification. The equipment provides a three-dimensional image of high contrast and resolution. The results of these microscopic observations can be used for both morphological (taxonomical) and ecological studies of microorganisms.

• MeSH
• Bacteria ultrastruktura   MeSH
• Eukaryota ultrastruktura   MeSH
• houby ultrastruktura   MeSH
• mikroskopie přístrojové vybavení   metody   MeSH
• světlo MeSH
• vylepšení obrazu MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Light microscopy (LM) approaches are commonly used to attain a description of the cell structure. Even though LM, if compared to electron microscopy (EM), represents a very fast approach, its resolution is, in principle, much lower than in the case of EM. To improve the LM resolution, computational methods based on removal of the image blur are frequently implemented in cell biology studies. One of the standard deblurring approaches is image restoration through deconvolution algorithms. Even though this method of mathematical remodeling of microscopically observed objects represents an efficient tool of current cell biology, it is legitimate to ask what the limits of its use are. We demonstrate that, in the specific case of the fluorescence mapping of active ribosomal genes in HeLa cell nucleoli, restoration generates a biased result. On restoration of model images, we demonstrate the difficulties of one of the most effective deconvolution algorithms during the restoration of ring-shaped fluorescent objects of a diameter comparable to the microscope resolution limit. In the case involving the mapping of nucleolar transcription in HeLa cells, not the restored fluorescence images, but rather the EM images show the true distribution of active ribosomal genes.

• MeSH
• buněčné jadérko metabolismus   MeSH
• elektronová mikroskopie metody   MeSH
• genetická transkripce MeSH
• HeLa buňky MeSH
• lidé MeSH
• mikroskopie metody   MeSH
• počítačové zpracování obrazu MeSH
• ribozomální DNA * MeSH
• ribozomy metabolismus   MeSH
• teoretické modely MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ribozomální DNA * MeSH