Fig. 1. System configuration. The top two insets are corresponding enlarged views of Sections 1 and 2, respectively. The right two insets are corresponding enlarged views of detection arrays of sCMOS1 and sCMOS2, respectively. The scanning coordinate system (F and S represent the fast axis and the slow axis, respectively) and imaging coordinate system are displayed.

Fig. 2. Image processing flow. (a) Pixel reassignment of X-LC and Y-LC results. (b) Merged pseudo-color strip image of imaging results by the X-LC (magenta) and Y-LC (green). (c) Registration results of three ROIs in (b). (d) Fusion result of the De2dir. The insets in (c) and (d) are the intensity profiles along the corresponding color lines. NCC, normalized cross correlation (green values in the top left corner). FWHM, full width at half maximum (blue values; in μm).

Fig. 3. Simulated results of a spoke-like sample. (a), (b) Raw images and corresponding spectra of the X-LC and Y-LC, respectively. (c), (d) Deconvolved images and corresponding spectra of the X-LC and Y-LC, respectively. (e) Fusion result and its spectrum of the cLIM. Region size, 4.7  μm×4.7  μm; pixel size, 13.5 nm.

Fig. 4. Comparison of different dual-direction fusion algorithms by imaging BPAE cells. (a) Pseudo-color-merged image of the X-LC (magenta) and Y-LC (green). Blue and white arrowheads indicate the differences between the signals contained in these two imaging results. (b) Fusion results by the Min_I,Max_I,Max_F, Prod, and De2dir. Dynamic ranges for displaying images are shown by the bars at the bottom right corners. (c) Corresponding enlarged views in (b). Intensity profiles along the colored lines are also shown at the bottom, including the maximum and minimum intensities, their ratios, and FWHM values. The yellow hollow and solid arrowheads indicate the signal distributions of the same microfilament in the different fusion algorithms. The scale bars in (a) and (c) are 50 μm and 2 μm, respectively.

Fig. 5. Lateral resolution measurement of the cLIM by imaging fluorescent beads. (a) Typical images of a fluorescent bead by the X-LC, Y-LC, and cLIM and the corresponding lateral resolution measurement by Gaussian fitting analyses. The statistical lateral resolution values are labeled (n=10). (b) Asymmetry of lateral resolution of the X-LC, Y-LC, and cLIM. Scanning pixel size, 0.077 μm.

Fig. 6. Imaging performance of the cLIM compared with LC and CM by imaging a 100-μm-thick β-actin-eGFP mouse kidney slice. (a) Imaging results of the LC, CM, and cLIM with numerically normalized maximum intensity value. (b) Enlarged views of the yellow box in (a) imaged by LC, CM, and cLIM. (c) Corresponding axial cuts imaged by LC, CM, and cLIM through the yellow dashed line in (b). (d) Enlarged views of the blue box in (b) imaged by LC, CM, and cLIM. (e) Intensity profiles along corresponding-colored lines in (d). (f) Grayscale histograms of the region of the yellow box in (a) imaged by the LC, CM, and cLIM. Mean intensities of the background are labeled in each figure. S, signal; B, background. Scale bars in (a)–(d) are 1 mm, 20 μm, 10 μm, and 2 μm, respectively.

Fig. 7. Imaging results of a 100-μm-thick β-actin-eGFP mouse tongue slice. (a) Structure tensor distribution of the cLIM imaging result. The insets display four types of intrinsic muscles of the mouse tongue. (b) Gradients along the X and Y directions in Region 1 indicated by the blue box in (a) of the corresponding imaging results using the X-LC, Y-LC, and cLIM. (c) Enlarged views of the corresponding imaging results of Region 2 indicated by the yellow rectangle in (a) using the X-LC, Y-LC, and cLIM. (d) Enlarged views of myofibril branching (yellow arrowheads) indicated by the blue rectangle in (c). (e) Enlarged views of sarcomeres indicated by the brown rectangle in (c). (f) Intensity profiles along corresponding-colored lines in (e) show the typical structures of the sarcomeres, including I-band, A-band, and Z-disk. SLM, superior longitudinal muscle; TM, transverse muscle; VM, vertical muscle; ILM, inferior longitudinal muscle.