Science Advances

Supplementary Materials

This PDF file includes:

  • Cell culture and treatment
  • BAT preparation
  • TPEF data acquisition
  • Segmentation algorithm for BAT image analysis
  • Calculation of heterogeneity index
  • fig. S1. Schematic of integrated analysis processing.
  • fig. S2. Raw data set for HFK cells under metabolic pathways of glycolysis or glutaminolysis, corresponding to representative images shown in Fig. 2.
  • fig. S3. Raw data set for HL-1 cells under chemical uncoupling, corresponding to representative images shown in Fig. 3.
  • fig. S4. Schematic of image segmentation of cytoplasm and lipids by taking into account both FAD fluorescence intensity and NADH bound fraction.
  • fig. S5. Ex vivo and in vivo raw data set for BAT under the treatment of cold activation, corresponding to representative images shown in Fig. 4.
  • fig. S6. Raw data set for C2C12 cells at various stages of β-oxidation, corresponding to representative images shown in Fig. 5 (A to F).
  • fig. S7. Fluorescence images of C2C12 cells with MitoTracker Green FM staining under different fatty acids supplement.
  • fig. S8. Raw data set for MSCs during lipogenesis, corresponding to representative images shown in Fig. 5 (G to L).
  • fig. S9. Optical readouts of C2C12 myoblasts in response to hypoxia, glucose starvation, or CCCP-induced chemical uncoupling.
  • fig. S10. Fluorescence images of C2C12 cells stained with tetramethylrhodamine ethylester (20 nM) under various treatments.
  • fig. S11. Classification of metabolic pathways using only one or two of the three optical metrics.
  • table S1. Individual heterogeneity index for each optical metric under different perturbation.
  • Legends for movies S1 to S3
  • List of abbreviations
  • References (98–100)

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Other Supplementary Material for this manuscript includes the following:

  • movie S1 (.avi format). Dynamic redox ratio maps of HFKs treated by normal media.
  • movie S2 (.avi format). Dynamic redox ratio maps of HFKs treated by hypoxia media.
  • movie S3 (.avi format). Dynamic redox ratio maps of HFKs treated by no-glucose media.

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