# Three-Dimension Resolution Enhanced Microscopy Based on Parallel Detection
## 3 Experimental Results
**(D)** The negative focus image at the focal plane. **(E)** The 3D negative focus pattern. (F) The local enlarge image of the single nanoparticle indicated in the dash frame of **(D)**. **(G)** The cross-section along the z axis of the same nanoparticle depicted in **(F)**. **(H,I)** are the images before Zernike correction corresponding to **(F,G)**. Figure 6: The comparison among 100 nm nanoparticle test results in CLSM, PRISM, and 3D-PRFED (\(\gamma\) = 0.7). **(A)** CLSM image at the focal plane. **(B)** PRISM image at the focal plane, **(C)** 3D-PRFED image at the focal plane. **(D–F)** are the cross-section view of a single nanoparticle in the dashed box region of **(A–C)**, respectively. **(G)** The transverse normalized profile along the white line in **(A–C)**. **(H)** The axial normalized profile along the dashed line in **(D–F)**. Scanning parameter: 6 μs/pixel, 30 nm transverse pixel size, 50 nm axial pixel size. In order to test the performance of 3D-PRFED applied in biological imaging, we imaged a slice of commercial nuclear pore complexes (NPCs) samples (Abberior Cells 3C NPC-START RED), as shown in Figure 7. The primary function of NPCs is as the key regulator of molecular traffic between the cytoplasm and the nucleus, and meanwhile this structure participates in a considerably broader range of cellular activities on both sides of the nuclear envelope. NPCs, among the largest proteinaceous assemblies in the cell, are made up of multiple copies of \(\sim\)30 different proteins called nucleoporins (Nups). The NPC in vertebrate consists of a \(\sim\)125 nm diameter core structure, which includes eight spokes in a radially symmetrical arrangement [24]. In the axial direction, 47 layers of NPC with 50 nm separation between consecutive layers have been acquired. Figure 7A-C are the results of CLSM, PRISM, and 3D-PRFED at 350 nm, which suggest that 3D-PRFED for the NPC imaging can obviously improve the resolution comparing with CLSM and PRISM. The normalized line profile in Figure 7G from Figure 7D-F, which are the local images come from the dash boxes of Figure 7A-C, suggest that two NPC structure can be obviously observed in 3D-PRFED but not in CLSM and PRISM. Figure 7H-J represent the cross-section view extracted along the white dashed line depicted in Figure 6A-C, which shows that the axial resolution can also be obviously improved in 3D-PRFED comparing with two other ways. As shown in Figure 7K which is the normalized line profile along the green line in Figure 7H-J, two NPC structures in different layers can be distinctly observed in 3D-PRFED but not in CLSM and PRISM. The NPC imaging experiments show that the excellent performance of 3D-PRFED can be well applied to biological imaging and other research projects. Figure 7: Nuclear pore complex results in CLSM, PRISM, and 3D-PRFED (\(\gamma\) = 0.7). (**A**) The CLSM image of this NPC sample at 350 nm plane. (**B**) The PRISM image of this NPC sample at 350 nm plane. (**C**) The 3D-PRFED image of this NPC sample at 350 nm plane. (**D–F**) are the local enlarged images in the dashed boxes of (**A–C**) respectively. (**G**) The normalized line profile along the green line in (**D–F**). (**H–J**) are the cross-section views along the dashed line in (**A–C**). (**K**) The line profile along the green line in (**H–J**). Scanning parameter: 6 μs/pixel, 30 nm transverse pixel size, 50 nm axial pixel size.