![]() The two last-mentioned techniques are particularly promising for resolving cellular migration and tissue rearrangements during quick morphogenetic events. High temporal and spatial resolutions can be achieved with lattice light-sheet microscopy, where a combination of ultrathin light sheets and structured illumination are used. For example, SCAPE (swept confocally-aligned planar excitation) microscopy offers more control over the viewing angle of the sample, while SVIM (selective volume illumination microscopy) dramatically increases acquisition times by dilating the light-sheet, at the expense of spatial resolution. Based on light-sheet illumination, novel and improved imaging techniques are continuously being developed. LSFM has been used to generate 3D time-lapse recordings of the complete embryonic morphogenesis of multiple model organisms. Besides confocal fluorescence microscopy, light-sheet fluorescence microscopy (LSFM) has become one of the preferred techniques for three-dimensional imaging of biological samples, owing to its fast acquisition times, excellent signal-to-noise ratios, high spatial resolutions, and low phototoxicity and photobleaching levels. Three-dimensional systems-such as model organisms, spheroids or organoids-are preferable, as they maintain physiological cell structures, neighborhood interactions, or mechanical extracellular properties, which have been recognized to play a role in regulating collective cellular migration. Studies of collective cell migration on 2D cell cultures only partially reflect the physiology and architecture of in vivo tissues. This phenomenon is generally known as collective cell migration, and it plays important roles in developmental processes, such as gastrulation or neural crest migration, as well as in wound closure and cancer invasion. In addition, our software includes functions to visualize the 3D vector fields in Paraview.Ĭellular migration in multi-cellular organisms often involves tissues or groups of cells that maintain stable or transient cell-cell contacts to preserve tissue integrity, sustain spatial patterning, or to enable the relocation of non-motile cells. Post-processing options include filtering and averaging of the resulting vector fields, extraction of velocity, divergence and collectiveness maps, simulation of pseudo-trajectories, and unit conversion. Currently, quickPIV offers efficient 2D and 3D PIV analyses featuring zero-normalized and normalized squared error cross-correlations, sub-pixel/voxel approximation, and multi-pass. The presented software addresses the need for a fast and open-source 3D PIV package in biological research. We show normalized squared error cross-correlation to be especially accurate in detecting translations in non-segmentable biological image data. Additionally, by applying quickPIV to three data sets of the embryogenesis of Tribolium castaneum, we obtained vector fields that recapitulate the migration movements of gastrulation, both in nuclear and actin-labeled embryos. The accuracy evaluation of our software on synthetic data agrees with the expected accuracies described in the literature. Our software is also faster than the fastest 2D PIV package in openPIV, written in C++. ![]() ![]() ![]() QuickPIV is three times faster than the Python implementation hosted in openPIV, both in 2D and 3D. Our software is focused on optimizing CPU performance and ensuring the robustness of the PIV analyses on biological data. This paper presents the implementation of an efficient 3D PIV package using the Julia programming language-quickPIV. ![]() Particle image velocimetry (PIV) is a robust and segmentation-free technique that is suitable for quantifying collective cellular migration on data sets with different labeling schemes. Dynamic 3D data sets of developing organisms allow for time-resolved quantitative analyses of morphogenetic changes in three dimensions, but require efficient and automatable analysis pipelines to tackle the resulting Terabytes of image data. The technical development of imaging techniques in life sciences has enabled the three-dimensional recording of living samples at increasing temporal resolutions. ![]()
0 Comments
Leave a Reply. |