Fig. 2: Characterization of quantitative light-field microscopy (QLFM). | Nature Communications

Fig. 2: Characterization of quantitative light-field microscopy (QLFM).

From: Computational optical sectioning with an incoherent multiscale scattering model for light-field microscopy

Fig. 2

a Illustrations of the SBR characterization experiment. We fabricated the scattering phantom with the mixture of 0.025% 2-μm fluorescence beads, 1% intralipid, and 1% agarose in a petri dish. Then we imaged the sample with a 40×/1.0NA water-immersion objective (Obj) at different penetration depths with WFM and LFM. b SBR curves of the fluorescence beads at different penetration depths in the intralipid-based tissue-mimicking phantom obtained by WFM, traditional LFM, QLFM with the multiscale model only, and QLFM with the multiscale scattering model. Data are presented as means ± SD. We chose 10 fluorescence beads with the highest fitting degrees for each block covering ~40 μm. QLFM shows ~20 dB improvement in the SBR over WFM, indicating an improved penetration depth in deep tissue. c Comparisons among the experimental PSF, the ideal PSF without aberrations, and the calibrated PSF of several selected angular components marked in the inset under the ×40/1.0 NA water-immersion objective. The calibrated wavefront of the system aberration estimated by our iterative phase-retrieval algorithm is shown on the left. d Lateral and axial resolution at different axial planes in traditional LFM and QLFM with 1 and 5 axially scanned steps, which is estimated by the FWHM of subdiffraction-limited fluorescence beads. Bar graphs are represented as means ± SD. For each block covering 10 μm, we chose 10 fluorescence beads with the highest fitting degrees for statistical analysis (one-way ANOVA with Tukey’s multiple comparisons test; the significance threshold was set at α = 0.05). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns not significant. e Reconstructed orthogonal slices of 0.5-μm-diameter fluorescence beads located at depths of 5 μm and 40 μm away from the focal plane by traditional LFM, QLFM with and without axial scanning of 5 steps. Cross-section profiles along the marked lines were plotted in the insets. Source data are provided as a Source data file. Scale bar: 3 μm.

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