Science Advances

Supplementary Materials

This PDF file includes:

  • fig. S1. Microfabrication steps of the electrocage chip for live-cell rotation.
  • fig. S2. Determination of the optimal voltage range of the electric field used to rotate live cells.
  • fig. S3. Cell rotation speed as a function of electric field frequency applied to the microelectrodes of the electrocage.
  • fig. S4. A live K562 cell incubated with 200-nm fluorescent polystyrene beads.
  • fig. S5. Photobleaching kinetics of the Hoechst 33342 dye in a live K562 cell.
  • fig. S6. Confocal micrographs showing enlarged mitochondrial structures in J774A.1 cells after treatment with 8-bromo-cAMP, a mitochondrial fission inhibitor.
  • fig. S7. A device used to simulate electric field conditions in the electrocage during cell rotation on bulk cell samples for assessing potential stress levels introduced by the electric field.
  • fig. S8. Testing potential cell stress caused by the exposure to high-frequency electric fields used in the electrocage for cell rotation using the device shown in
  • fig. S9. Assessing potential changes in cellular morphology as a result of exposure to high-frequency electric fields via imaging.
  • fig. S10. Imaging system setup and implementation.
  • fig. S11. A diagram of the overall LCCT system design and control.
  • fig. S12. Piezo scanning, control, and synchronization time diagrams (green, objective scanning voltage; red, triggering pulse; and yellow, acquisition exposure control waveform).
  • fig. S13. Comparison of the reconstruction of the various registration methods.
  • fig. S14. Computational workflow used in the modified SIRT and blind deconvolution SIRT methods of volumetric image reconstruction.
  • fig. S15. Performance demonstration of four different volumetric reconstruction approaches used in the study.
  • fig. S16. The principle of sinogram generation.
  • fig. S17. Examples of detector and slice sinograms generated from simulated
    projection images of two beads moving in circular, distortion-free trajectories.
  • fig. S18. Description of the GeoFit algorithm computational pipeline, which estimates and corrects in-plane projection perturbations—Lateral shift and in-plane orientation changes of the rotation axis.
  • fig. S19. Correction of in-plane perturbations of rotation using the GeoFit algorithm.
  • fig. S20. Slice of a volumetric image reconstructed using raw/uncorrected data (left) and after correction using the GeoFit algorithm (right).
  • fig. S21. Pipeline description of the FixPP algorithm to correct out-of-plane perturbations.
  • fig. S22. Reconstruction of simulated Shepp-Logan data with 10° axis elevation using the FixPP algorithm.
  • Legends for movies S1 to S17

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Other Supplementary Material for this manuscript includes the following:

  • movie S1 (.avi format). Raw PP images of a live human myelogenous leukemia (K562 cell line) cell rotating in the electrocage.
  • movie S2 (.avi format). Raw PP images of a live K562 cell with internalized 200-nm fluorescent beads rotating in the electrocage.
  • movie S3 (.mpg format). Reconstructed 3D volumetric image (surface rendering) of the K562 cell shown in movie S2.
  • movie S4 (.mpg format). Comparison between the confocal and LCCT imaging modalities.
  • movie S5 (.mpg format). Surface rendering of a reconstructed 3D volumetric image of a live K562 cell with stained nucleus (blue-green) and mitochondria (red-yellow).
  • movie S6 (.mpg format). Nuclear feature segmentation of a reconstructed 3D volumetric image of a live K562 cell.
  • movie S7 (.mpg format). Mitochondrial feature segmentation of a reconstructed 3D volumetric image of the same cell as shown in movie S4.
  • movie S8 (.mpg format). Overlay of nuclear (green) and mitochondrial (red) feature segmentation results and their corresponding MIP renderings shown in movies S4 and S5.
  • movie S9 (.mpg format). MIP and separate slices of the reconstructed 3D volumetric images of mitochondria in the same cell shown in movies S5 to S8.
  • movie S10 (.mpg format). Mitochondrial fluorescence intensity and segmentation results using the Niblack local threshold approach.
  • movie S11 (.mpg format). 3D view of mitochondrial segmentation overlaid with fluorescence intensity (both surface renderings).
  • movie S12 (.mpg format). A representative example of mitochondrial segmentation in 3D illustrated as a small ROI from movie S11.
  • movie S13 (.mpg format). 3D rendering and Z-stack of the mitochondrial network in a fixed K562 cell imaged using confocal microscopy.
  • movie S14 (.mpg format). Mitochondrial fluorescence intensity and segmentation results of the cell shown in movie S13.
  • movie S15 (.mpg format). Fluorescence intensity and segmentation results of the nucleoli in a live K562 cell.
  • movie S16 (.mpg format). Maximum intensity renderings of reconstructed 3D images of an untreated (left) and treated (right) J774A.1 cell line (mouse macrophage cell) with the mitochondrial fission inhibitor 8-bromo-cAMP.
  • movie S17 (.mpg format). Surface rendering of a reconstructed 3D image of a live J774A.1 cell.

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