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Holograms carrying both amplitude and phase are acquired at multiple illumination angles, and the inverse problem is solved to produce a 3D RI map. This method is called optical diffraction tomography. It is the optical counterpart of CT reconstruction.
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The illumination is low-power visible light. There is no fluorescence excitation and nothing to photobleach, so the photodamage burden in long time-lapse experiments is structurally lower than with fluorescence imaging.
The achievable duration is not unlimited. It depends on cell type and culture conditions, and we recommend confirming it against your own protocol in a demo or pilot experiment.
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Lateral resolution is at the diffraction-limited scale. Axial resolution is lower than lateral because of the missing cone, a physical property of optical diffraction tomography.
Resolution and imaging depth differ by model. Refer to the specification sheet of each HT-X1 series imaging system, or contact us for model-specific figures.
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Multiple scattering increases with sample thickness and density, and this limits reconstruction in every optical tomography method. For spheroids and organoids, the outcome depends on their size and density, so it has to be measured rather than predicted.
If you are working with thick samples, contact us to arrange a feasibility test with your own sample.
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Holotomography complements fluorescence rather than replacing it. Fluorescence gives molecular specificity. Holotomography covers the cases where labeling is not possible or not desirable: long-term tracking, measuring the baseline state before any label is introduced, primary cells and stem cells, and experiments where you want to keep fluorescence channels free for other targets.
In practice the two are used together. HT-X1 series imaging systems support correlative imaging, which gives quantitative structural measurements and molecular specificity on the same cell.
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Three points distinguish the approach:
- 3D quantitative tomography versus 2D phase. A 2D quantitative phase image integrates optical path difference along the beam, so thickness and density are mixed in one value. A 3D RI tomogram separates them.
- Workflow completeness. Environmental control for live cells, multi-well acquisition, long unattended time-lapse, and analysis software are provided as one product family.
- Published work. Peer-reviewed publications using Tomocube systems are listed on our Publications page, and references for your specific application are available on request.
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Multi-well automated acquisition is supported. Actual throughput depends on how the experiment is designed: fields of view per well, z-range, and time interval. Share your screen design with us and we will calculate the expected acquisition time.
Holotomography data supports label-free phenotypic profiling across conditions.
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All Tomocube products are for Research Use Only. They are not sold for diagnostic or therapeutic use. Research involving clinical specimens must proceed under your institution's own approvals, including IRB review where applicable.