scholarly journals Micro-CT for Biological and Biomedical Studies: A Comparison of Imaging Techniques

2021 ◽  
Vol 7 (9) ◽  
pp. 172
Author(s):  
Kleoniki Keklikoglou ◽  
Christos Arvanitidis ◽  
Georgios Chatzigeorgiou ◽  
Eva Chatzinikolaou ◽  
Efstratios Karagiannidis ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Imaging Techniques ◽  
Laser Scanning Microscopy ◽  
Three Dimensions ◽  
Morphological Data ◽  
Confocal Laser ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
Non Destructive ◽  
Great Resolution

Several imaging techniques are used in biological and biomedical studies. Micro-computed tomography (micro-CT) is a non-destructive imaging technique that allows the rapid digitisation of internal and external structures of a sample in three dimensions and with great resolution. In this review, the strengths and weaknesses of some common imaging techniques applied in biological and biomedical fields, such as optical microscopy, confocal laser scanning microscopy, and scanning electron microscopy, are presented and compared with the micro-CT technique through five use cases. Finally, the ability of micro-CT to create non-destructively 3D anatomical and morphological data in sub-micron resolution and the necessity to develop complementary methods with other imaging techniques, in order to overcome limitations caused by each technique, is emphasised.

A Brief Review of Visualization Techniques for Nerve Tissue Engineering Applications

2010 ◽  
Vol 7 ◽  
pp. 81-99 ◽  
Author(s):  
Ning Zhu ◽  
Xiong Biao Chen ◽  
Dean Chapman
Keyword(s):  
Tissue Engineering ◽  
Laser Scanning ◽  
Structural Features ◽  
Laser Scanning Microscopy ◽  
Nerve Tissue ◽  
Confocal Laser ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
Nerve Tissue Engineering ◽  
Visualization Techniques

In nerve tissue engineering, scaffolds act as carriers for cells and biochemical factors and as constructs providing appropriate mechanical conditions. During nerve regeneration, new tissue grows into the scaffolds, which degrade gradually. To optimize this process, researchers must study and analyze various morphological and structural features of the scaffolds, the ingrowth of nerve tissue, and scaffold degradation. Therefore, visualization of the scaffolds as well as the generated nerve tissue is essential, yet challenging Visualization techniques currently used in nerve tissue engineering include electron microscopy, confocal laser scanning microscopy (CLSM), and micro-computed tomography (micro-CT or μCT). Synchrotron-based micro-CT (SRμCT) is an emerging and promising technique, drawing considerable recent attention. Here, we review typical applications of these visualization techniques in nerve tissue engineering. The promise, feasibility, and challenges of SRμCT as a visualization technique applied to nerve tissue engineering are also discussed.

scholarly journals Protocol for rapid clearing and staining of fixed Arabidopsis ovules for improved imaging by confocal laser scanning microscopy

Plant Methods ◽  
2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Rachele Tofanelli ◽  
Athul Vijayan ◽  
Sebastian Scholz ◽  
Kay Schneitz
Keyword(s):  
Laser Scanning ◽  
Developmental Stages ◽  
Imaging Techniques ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Imaging Data ◽  
Model Species ◽  
Cellular Models ◽  
Cellular Resolution

Abstract Background A salient topic in developmental biology relates to the molecular and genetic mechanisms that underlie tissue morphogenesis. Modern quantitative approaches to this central question frequently involve digital cellular models of the organ or tissue under study. The ovules of the model species Arabidopsis thaliana have long been established as a model system for the study of organogenesis in plants. While ovule development in Arabidopsis can be followed by a variety of different imaging techniques, no experimental strategy presently exists that enables an easy and straightforward investigation of the morphology of internal tissues of the ovule with cellular resolution. Results We developed a protocol for rapid and robust confocal microscopy of fixed Arabidopsis ovules of all stages. The method combines clearing of fixed ovules in ClearSee solution with marking the cell outline using the cell wall stain SCRI Renaissance 2200 and the nuclei with the stain TO-PRO-3 iodide. We further improved the microscopy by employing a homogenous immersion system aimed at minimizing refractive index differences. The method allows complete inspection of the cellular architecture even deep within the ovule. Using the new protocol we were able to generate digital three-dimensional models of ovules of various stages. Conclusions The protocol enables the quick and reproducible imaging of fixed Arabidopsis ovules of all developmental stages. From the imaging data three-dimensional digital ovule models with cellular resolution can be rapidly generated using image analysis software, for example MorphographX. Such digital models will provide the foundation for a future quantitative analysis of ovule morphogenesis in a model species.

scholarly journals Reconstruction of Axial Tomographic High Resolution Data from Confocal Fluorescence Microscopy: A Method for Improving 3D FISH Images

2000 ◽  
Vol 20 (1) ◽  
pp. 7-15 ◽  
Author(s):  
R. Heintzmann ◽  
G. Kreth ◽  
C. Cremer
Keyword(s):  
High Resolution ◽  
Laser Scanning ◽  
Axial Direction ◽  
Laser Scanning Microscopy ◽  
Three Dimensions ◽  
Confocal Laser ◽  
Data Sets ◽  
Data Set ◽  
High Resolution Data ◽  
3D Data

Fluorescent confocal laser scanning microscopy allows an improved imaging of microscopic objects in three dimensions. However, the resolution along the axial direction is three times worse than the resolution in lateral directions. A method to overcome this axial limitation is tilting the object under the microscope, in a way that the direction of the optical axis points into different directions relative to the sample. A new technique for a simultaneous reconstruction from a number of such axial tomographic confocal data sets was developed and used for high resolution reconstruction of 3D‐data both from experimental and virtual microscopic data sets. The reconstructed images have a highly improved 3D resolution, which is comparable to the lateral resolution of a single deconvolved data set. Axial tomographic imaging in combination with simultaneous data reconstruction also opens the possibility for a more precise quantification of 3D data. The color images of this publication can be accessed from http://www.esacp.org/acp/2000/20‐1/heintzmann.htm. At this web address an interactive 3D viewer is additionally provided for browsing the 3D data. This java applet displays three orthogonal slices of the data set which are dynamically updated by user mouse clicks or keystrokes.

scholarly journals The second axillary in Hymenoptera

2014 ◽  
Author(s):  
István Mikó ◽  
Andy R Deans
Keyword(s):  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
3D Visualization ◽  
Laser Scanning Microscopy ◽  
Morphological Data ◽  
Confocal Laser ◽  
Wing Membrane ◽  
Future Directions ◽  
Wing Base ◽  
Visualization Techniques

The wing base of basal hymenopterans (Insecta) have never been properly described perhaps due to the difficulties of its visualization and understanding the 3D relationships between wing base components. Novel 3D visualization techniques such as microCT and Confocal Laser Scanning Microscopy (CLSM) allow us to provide easily digestible morphological data. The wing base of four basal Hymenoptera and 10 apocritan species have been imaged with CLSM and dissected under a stereomicroscope. The second axillary is composed of two sclerites (on on the dorsal wing membrane and one on the ventral in Macroxyela, Xyela and Athalia whereas it is represented by a single sclerite traversing the wing in other Hymenoptera. Consequences related to this observation as well are drawn and future directions in Hymenoptera wing base studies are provided.

Confocal laser scanning microscopy and Raman imagery of the late Neoproterozoic Chichkan microbiota of South Kazakhstan

2010 ◽  
Vol 84 (3) ◽  
pp. 402-416 ◽  
Author(s):  
J. William Schopf ◽  
Anatoliy B. Kudryavtsev ◽  
Vladimir N. Sergeev
Keyword(s):  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Three Dimensions ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
Depth Study ◽  
Late Neoproterozoic

Precambrian microbiotas, such as that permineralized in bedded and stromatolitic cherts of the late Neoproterozoic, 750- to 800-Ma-old, Chichkan Formation of South Kazakhstan, have traditionally been studied by optical microscopy only. Such studies, however, are incapable of documenting accurately either the three-dimensional morphology of such fossils or their chemical composition and that of their embedding minerals. As shown here by analyses of fossils of the Chichkan Lagerstätte, the solution to these long-standing problems is provided by two techniques recently introduced to paleontology: confocal laser scanning microscopy (CLSM) and Raman imagery. The two techniques are used together to characterize, in situ and at micron-scale resolution, the cellular and organismal morphology of the thin section-embedded organic-walled Chichkan fossils. In addition, Raman imagery is used to analyze the molecular-structural composition of the carbonaceous fossils and of their embedding mineral matrix, identify the composition of intracellular inclusions, and quantitatively assess the geochemical maturity of the Chichkan organic matter.CLSM and Raman imagery are both broadly applicable to the study of fossils, whether megascopic or microscopic and regardless of mode of preservation, and both are non-intrusive and non-destructive, factors that permit their use for analyses of archived specimens. They are especially useful for the study of microscopic fossils, as is demonstrated in this first in-depth study of diverse taxa of a single Precambrian microbiota for which they provide information in three dimensions at high spatial resolution about their organismal morphology, cellular anatomy, kerogenous composition, mode of preservation, and taphonomy and fidelity of preservation.

scholarly journals Morphological aspects of self-repair of lesions caused by internal growth stresses in stems of Aristolochia macrophylla and Aristolochia ringens

2010 ◽  
Vol 277 (1691) ◽  
pp. 2113-2120 ◽  
Author(s):  
Sebastian Busch ◽  
Robin Seidel ◽  
Olga Speck ◽  
Thomas Speck
Keyword(s):  
Cell Wall ◽  
Laser Scanning ◽  
Early Stage ◽  
Morphological Changes ◽  
Secondary Growth ◽  
Laser Scanning Microscopy ◽  
Morphological Data ◽  
Confocal Laser ◽  
Growth Stresses ◽  
Self Repair

This study reveals in detail the mechanism of self-repair during secondary growth in the vines Aristolochia macrophylla and Aristolochia ringens based on morphological data. For a comprehensive understanding of the underlying mechanisms during the self-repair of lesions in the sclerenchymatous cylinder of the stem, which are caused by internal growth stresses, a classification of morphological changes in the cells involved in the repair process is required. In an early stage of self-repair, we observed morphological changes as a mere extension of the turgescent cortex cells surrounding the lesion, whereby the cell wall extends locally through visco-elastic/plastic deformation without observable cell wall synthesis. Later stages involve typical cell growth and cell division. Several successive phases of self-repair were investigated by light microscopy of stained samples and confocal laser-scanning microscopy in fluorescence mode. The results indicate that A. macrophylla and A. ringens respond to lesions caused by internal growth stresses with a sophisticated self-repair mechanism comprising several phases of different repair modes.

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