Fig. 1. (a) The schematic of the PM3 technique. LED port, light-emitting diode output port; Laser port, 532 nm CW laser output port; SLM, spatial light modulator; Block, an aperture; CCD, charge-coupled device. (b) The schematic of the working principle for PM3, whereby the scattered or emitted fluorescence light from the out-of-focus position is collimated and deflected by the modulated phase (represented by the green curved line). Images from multiple depths are then focused onto different areas of the CCD camera. (c) The phase pattern used in the experiment to capture the images from three different depths. (d) Enlarged view of the image outlined by a red dashed rectangle in (c). The section with a red solid rectangle represents the phase pattern used for the scattered or emitted fluorescence light from depth z1, while the green and blue sections correspond to the depths z2 and z3, respectively. (e) The schematic of the images from depths z1,z2,z3 on the CCD camera. (f) The phase pattern used to capture images from 10 different depths. (g) Enlarged view of the image outlined by a red dashed rectangle in (f). (h) The schematic of the CCD plane with the images from 10 different depths. (i) The fit axial intensity profiles of a 1 μm polymer bead measured with 10 foci. (j) The axial position of each focal plane.

Fig. 2. (a) Schematic of wide-range volumetric imaging for the Brownian motion of PS beads in water by PM3. The gray long rectangle represents the water channel, and the blue rectangle indicates the imaging volume consisting of 10 focal planes generated by PM3. When the beads move along the channel, the imaging volume of PM3 also dynamically shifts along the z-axis to capture the 3D trajectory of PS beads in water over a considerable distance. (b) The beads image captured at each focal plane (Δz=18  μm) within the imaging volume. Scalebar: 20 μm. (c) Snapshot images depicting the 3D distribution of beads at different axial positions in the imaging volume. The size of the volume is 78  μm×78  μm×162  μm. (d), (e) The trajectory of the 2 PS beads [white dotted rectangle in (c)] observed both in the water channel in (a) and within the imaging volume. (f)–(h) Dynamic fluorescence 3D images of 6 μm fluorescent beads in water. Scalebar: 50 μm. Image volume: 208  μm×208  μm×200  μm. 67 ms acquisition time for obtaining one 3D volume (15 Hz).

Fig. 3. (a) The enlarged phase pattern used to capture images from three different tissue depths with the same imaging interval. The region enclosed by a yellow dashed rectangle signifies the modulation phase for the scattered light from z=30  μm. (b)–(d) The snapshot images of blood flows (yellow dashed line) and red blood cell (red dashed ellipse) at depths of 0 μm, 30 μm, and 60 μm, respectively. Scale bar: 20 μm. (e) The enlarged phase pattern employed to obtain images from three different depths with varying imaging intervals. The area surrounded by a yellow dashed rectangle indicates the modulation phase for the scattered light from z=10  μm. (f)–(h) The snapshot images of blood flows (yellow dashed line) and red blood cells (red dashed ellipse) captured by PM3 at depths of 0 μm, 10 μm, and 60 μm, respectively. Scale bar: 20 μm.

Fig. 4. (a)–(d) Segment images for 10, 30, 60 pies, and an undivided pupil, respectively. (e)–(h) Lateral spot images corresponding to the segment images in (a)–(d) accordingly. (i)–(l) Axial spot images corresponding to the images in (a)–(d) accordingly. (m), (n) Lateral and axial intensity distributions related to different segment schemes in (a)–(d), respectively.

Fig. 5. Snapshot image of blood flow in zebrafish trunk obtained by PM3, where three sub-windows represent the images from three depths with the same imaging interval.

Fig. 6. Snapshot image of blood flow in the same zebrafish trunk captured by PM3, in which three sub-windows represent the images from three depths with different imaging intervals.

Fig. 7. Snapshot 3D image of Brownian motion of fluorescent beads in water.