TL;DR
4Dia is an ImageJ/Fiji macro tool that automates the alignment of 3D live microscopy images over time, addressing challenges of misalignment due to biological and technical factors in long-term imaging.
Contribution
The paper introduces 4Dia, a versatile, user-friendly macro for automated 4D image alignment applicable to various tissue samples and imaging conditions.
Findings
Successfully aligns large 3D timelapse datasets
Compatible with any tissue sample and imaging setup
Improves accuracy of long-term live imaging analysis
Abstract
Recent advances in microscopy enable three-dimensional live imaging at a high resolution. Long-term live imaging of a multicellular organism requires immobilization of the organism under stable physiological conditions. Despite proper immobilization, challenges remain within live imaging data analysis due to other intrinsic and extrinsic dynamics, which can result in misalignments of an image series over time. 4Dia, an ImageJ/Fiji macro script, aligns 3D timelapse images through Z-stacks as well as over time using any user selected channel. 4Dia can be used for essentially any tissue sample with no limit on the size of Z-stack or the number of timepoints.
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