New evidence of the Hoyle-like structure in 16O.

An experiment of 12C(16O,16O → 4α)12C was performed at a beam energy of 96 MeV. A large number of 4-α events were recorded in coincidence and with full particle identification (PID). This was made possible by employing a series of silicon-strip-based telescopes that provided excellent position and energy resolutions. Four narrow resonances just above the 15.1 MeV state were firmly identified in the α + 12C(7.65 MeV; Hoyle state) decay channel. Combined with the theoretical predictions, these resonant states provide new evidence for the predicted possible Hoyle-like structure in 16O above the 4-α separation threshold. Some very high-lying 4-α resonant states have also been observed and need to be further investigated.

Resolution and uniformity improvement of parallel confocal microscopy based on microlens arrays and a spatial light modulator.

In traditional fluorescence microscopy, it is hard to achieve a large uniform imaging field with high resolution. In this manuscript, we developed a confocal fluorescence microscope combining the microlens array with spatial light modulator to address this issue. In our system, a multi-spot array generated by a spatial light modulator passes through the microlens array to form an optical probe array. Then multi-spot adaptive pixel-reassignment method for image scanning microscopy (MAPR-ISM) will be introduced in this parallelized imaging to improve spatial resolution. To generate a uniform image, we employ an optimized double weighted Gerchberg-Saxton algorithm (ODWGS) using signal feedback from the camera. We have built a prototype system with a FOV of 3.5 mm × 3.5 mm illuminated by 2500 confocal points. The system provides a lateral resolution of ∼0.82 µm with ∼1.6 times resolution enhancement after ISM processing. And the nonuniformity across the whole imaging field is 3%. Experimental results of fluorescent beads, mouse brain slices and melanoma slices are presented to validate the applicability and effectiveness of our system.

Multicolor high-resolution whole-brain imaging for acquiring and comparing the brain-wide distributions of type-specific and projection-specific neurons with anatomical annotation in the same brain.

Visualizing the relationships and interactions among different biological components in the whole brain is crucial to our understanding of brain structures and functions. However, an automatic multicolor whole-brain imaging technique is still lacking. Here, we developed a multicolor wide-field large-volume tomography (multicolor WVT) to simultaneously acquire fluorescent signals in blue, green, and red channels in the whole brain. To facilitate the segmentation of brain regions and anatomical annotation, we used 4', 6-diamidino-2-phenylindole (DAPI) to provide cytoarchitecture through real-time counterstaining. We optimized the imaging planes and modes of three channels to overcome the axial chromatic aberration of the illumination path and avoid the crosstalk from DAPI to the green channel without the modification of system configuration. We also developed an automatic contour recognition algorithm based on DAPI-staining cytoarchitecture to shorten data acquisition time and reduce data redundancy. To demonstrate the potential of our system in deciphering the relationship of the multiple components of neural circuits, we acquired and quantified the brain-wide distributions of cholinergic neurons and input of ventral Caudoputamen (CP) with the anatomical annotation in the same brain. We further identified the cholinergic type of upstream neurons projecting to CP through the triple-color collocated analysis and quantified its proportions in the two brain-wide distributions. Both accounted for 0.22%, implying CP might be modulated by non-cholinergic neurons. Our method provides a new research tool for studying the different biological components in the same organ and potentially facilitates the understanding of the processing mechanism of neural circuits and other biological activities.

Single-scan HiLo with line-illumination strategy for optical section imaging of thick tissues.

Optical sectioning has been widely employed for inhibiting out-of-focus backgrounds in three-dimensional (3D) imaging of biological samples. However, point scanning imaging or multiple acquisitions for wide-field optical sectioning in epi-illumination microscopy remains time-consuming for large-scale imaging. In this paper, we propose a single-scan optical sectioning method based on the hybrid illumination (HiLo) algorithm with a line-illumination strategy. Our method combines HiLo background inhibition with confocal slit detection. It thereby offers a higher optical sectioning capability than wide-field HiLo and line-confocal imaging without extra modulation and multiple data acquisition. To demonstrate the optical-sectioning capability of our system, we imaged a thin fluorescent plane and different fluorescence-labeled mouse tissue. Our method shows an excellent background inhibition in thick tissue and thus potentially provides an alternative tool for 3D imaging of large-scale biological tissue.