Serial block face SEM visualization of unusual plant nuclear tubular extensions in a carnivorous plant (Utricularia, Lentibulariaceae)

Płachno, Bartosz J.; Świątek, Piotr; Jobson, Richard W.; Małota, Karol; Brutkowski, Wojciech

doi:10.1093/aob/mcx042

Cited by 24 publications

(15 citation statements)

References 31 publications

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“…A promising application of the electron microscopy to plant specimens would be the correlative light and electron microscopy in which the light microscopy and the TEM are used in a complementary manner [9]. Another perspective is the serial block face SEM in which consecutive SEM images are integrated to reconstruct a large volume with dimensions of hundreds of microns [10,11]. Although the electron microscopy is quite useful for the high-resolution observation, it is time consuming and requires a laborious preparation of ultrathin specimens with thicknesses of around 100 nm, due to a low penetration power of electron beams.…”

Section: Introductionmentioning

confidence: 99%

Visualization of internal 3D structure of small live seed on germination by laboratory-based X-ray microscopy with phase contrast computed tomography

et al. 2020

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Background: The visualization of internal 3D-structure of tissues at micron resolutions without staining by contrast reagents is desirable in plant researches, and it can be achieved by an X-ray computed tomography (CT) with a phase-retrieval technique. Recently, a laboratory-based X-ray microscope adopting the phase contrast CT was developed as a powerful tool for the observation of weakly absorbing biological samples. Here we report the observation of unstained pansy seeds using the laboratory-based X-ray phase-contrast CT. Results: A live pansy seed within 2 mm in size was simply mounted inside a plastic tube and irradiated by in-house X-rays to collect projection images using a laboratory-based X-ray microscope. The phase-retrieval technique was applied to enhance contrasts in the projection images. In addition to a dry seed, wet seeds on germination with the poorer contrasts were tried. The phase-retrieved tomograms from both the dry and the wet seeds revealed a cellular level of spatial resolutions that were enough to resolve cells in the seeds, and provided enough contrasts to delineate the boundary of embryos manually. The manual segmentation allowed a 3D rendering of embryos at three different stages in the germination, which visualized an overall morphological change of the embryo upon germination as well as a spatial arrangement of cells inside the embryo. Conclusions: Our results confirmed an availability of the laboratory-based X-ray phase-contrast CT for a 3D-structural study on the development of small seeds. The present method may provide a unique way to observe live plant tissues at micron resolutions without structural perturbations due to the sample preparation.

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Section: Introductionmentioning

confidence: 99%

Visualization of internal 3D structure of small live seed on germination by laboratory-based X-ray microscopy with phase contrast computed tomography

et al. 2020

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“…To visualize the morphology and distribution of the mitochondria within a germ-line cyst, we prepared three-dimensional ultrastructural reconstructions using the SBEM technique. is technique is based on installation of the ultramicrotome inside the SEM chamber, which enables sequential ultrastructural micrographs of analyzed structures to be obtained with EM resolution, which is the basis for the 3D visualizations [64,65]. It is also worth mentioning that we did not concentrate solely on a fragment of a cell (as in the Bb that has been reconstructed in T. domestica oocytes) [31] or even at a single cell (as in several other studies) [64,66], but we reconstructed the entire selected compartments of a germ-line cyst, i.e.…”

Section: Introductionmentioning

confidence: 99%

Morphology of Mitochondria in Syncytial Annelid Female Germ-Line Cyst Visualized by Serial Block-Face SEM

Urbisz

Student

Śliwińska

et al. 2020

International Journal of Cell Biology

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Mitochondria change their morphology and distribution depending on the metabolism and functional state of a cell. Here, we analyzed the mitochondria and selected structures in female germ-line cysts in a representative of clitellate annelids – the white worm Enchytraeus albidus in which each germ cell has one cytoplasmic bridge that connects it to a common cytoplasmic mass. Using serial block-face scanning electron microscopy (SBEM), we prepared three-dimensional ultrastructural reconstructions of the entire selected compartments of a cyst at the advanced stage of oogenesis, i.e. the nurse cell, cytophore, and cytoplasmic bridges of all 16 cells (15 nurse cells and oocyte). We revealed extensive mitochondrial networks in the nurse cells, cytophore and mitochondria that pass through the cytoplasmic bridges, which indicates that a mitochondrial network can extend throughout the entire cyst. The dynamic hyperfusion state was suggested for such mitochondrial aggregations. We measured the mitochondria distribution and revealed their polarized distribution in the nurse cells and more abundant accumulation within the cytophore compared to the nurse cell. A close association of mitochondrial networks with dispersed nuage material, which seems to be the structural equivalent of a Balbiani body, not described in clitellate annelids so far, was also revealed.

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“…On the subcellular level, studies have utilised SBFSEM to examine the ultrastructure of many cell components. A recent study examined the morphology of carnivorous plant nuclei in order to examine novel membrane extensions (Plachno et al, 2017). Sub-nuclear structures such as individual chromosomes (Chen et al, 2017) and the mitotic spindle (Nixon et al, 2017) have been imaged and modelled.…”

Section: Serial Block Face Scanning Electron Microscopymentioning

confidence: 99%

Serial block face scanning electron microscopy in cell biology: Applications and technology

Smith

Starborg

2019

Tissue and Cell

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Three-dimensional electron microscopy (3DEM) is an imaging field containing several powerful modalities such as serial section transmission electron microscopy and electron tomography. However, large-scale 3D studies of biological ultrastructure on a cellular scale have historically been hampered by the difficulty of available techniques. Serial block face scanning electron microscopy (SBFSEM) is a 3DEM technique, developed in 2004, which has greatly increased the reliability, availability and throughput of 3DEM. SBFSEM allows for 3D imaging at resolutions high enough to resolve membranes and small vesicles whilst having the capability to collect data with a large field of view. Since its introduction it has become a major tool for ultrastructural investigation and has been applied in the study of many biological fields, such as connectomics, cellular and matrix biology. In this review, we will discuss biological SBFSEM from a technical standpoint, with a focus on cellular applications and also subsequent image analysis techniques.

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Serial block face SEM visualization of unusual plant nuclear tubular extensions in a carnivorous plant (Utricularia, Lentibulariaceae)

Cited by 24 publications

References 31 publications

Visualization of internal 3D structure of small live seed on germination by laboratory-based X-ray microscopy with phase contrast computed tomography

Visualization of internal 3D structure of small live seed on germination by laboratory-based X-ray microscopy with phase contrast computed tomography

Morphology of Mitochondria in Syncytial Annelid Female Germ-Line Cyst Visualized by Serial Block-Face SEM

Serial block face scanning electron microscopy in cell biology: Applications and technology

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