All confocal images that were used in the paper were taken with a Zeiss LSM 780 microscope, using 63x objective and 1.6x zoom (1024x1024). Each Z section was 0.37 um apart and each image consists of 20-40 z sections.
Open a confocal image in Imaris.
Create a new Surface.
Choose "Skip automatic creation, edit manually".
Choose "Source Channel" (e.g., Channel 1 - Green).
Choose "Smooth Detail" – for most of the images in the paper, 0.165 um was used.
Choose "Thresholding" – for most of the images in the paper, "Absolute Intensity" was used and manually adjusted.
Filter using "Number of Voxels" if needed.
Finish all creation steps.
Return to step 2 for the next channel if needed.
Choose different colors for each surface created if needed.
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How to cite:Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
- Chung, S and Andrew, D(2020). 3D reconstruction of SGs. Bio-protocol Preprint. bio-protocol.org/prep358.
- Chung, S., Kim, S. and Andrew, D. J.(2017). Uncoupling apical constriction from tissue invagination. eLife. DOI: 10.7554/eLife.22235
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