Synthesis, preclinical evaluation and pilot clinical translation of [68Ga]Ga-PMD22, a novel nanobody PET probe targeting CLDN18.2 of gastrointestinal cancer.

PURPOSE: Claudin18.2 (CLDN18.2) is a novel target for diagnosis and therapy of gastrointestinal cancer. This study aimed to evaluate the safety and feasibility of a novel CLDN18.2-targeted nanobody, PMD22, labeled with gallium-68 ([68Ga]Ga), for detecting CLDN18.2 expression in patients with gastrointestinal cancer using PET/CT imaging. METHODS: [68Ga]Ga-PMD22 was synthesized based on the nanobody, and its cell binding properties were assayed. Preclinical pharmacokinetics were determined in CLDN18.2-positive xenografts using microPET/CT. Effective dosimetry of [68Ga]Ga-PMD22 was evaluated in 5 gastrointestinal cancer patients, and PET/CT imaging of [68Ga]Ga-PMD22 and [18F]FDG were performed head-to-head in 16 gastrointestinal cancer patients. Pathological tissues were obtained for CLDN18.2 immunohistochemical (IHC) staining and comparative analysis with PET/CT findings. RESULTS: Cell binding assay showed that [68Ga]Ga-PMD22 had a higher binding ability to AGSCLDN18.2 and BGC823CLDN18.2 cells than to AGS and BGC823 cells (p < 0.001). MicroPET/CT images showed that [68Ga]Ga-PMD22 rapidly accumulated in AGSCLDN18.2 and BGC823CLDN18.2 tumors, and high contrast tumor to background imaging was clearly observed. In the pilot study, the effective dose of [68Ga]Ga-PMD22 was 1.68E-02 ± 1.45E-02 mSv/MBq, and the CLDN18.2 IHC staining result was highly correlated with the SUVmax/BKGstomach of [68Ga]Ga-PMD22 (rs = 0.848, p < 0.01). CONCLUSION: A novel [68Ga]Ga-labeled nanobody probe targeting CLDN18.2, [68Ga]Ga-PMD22, was established and preliminarily proved to be safe and effective in revealing CLDN18.2-positive gastrointestinal cancer, providing a basis for the clinical translation of the agent. CLINICAL TRIAL REGISTRATION: This study was registered on the ClinicalTrials.gov (NCT05937919).

DiffDomain enables identification of structurally reorganized topologically associating domains.

Topologically associating domains (TADs) are critical structural units in three-dimensional genome organization of mammalian genome. Dynamic reorganizations of TADs between health and disease states are associated with essential genome functions. However, computational methods for identifying reorganized TADs are still in the early stages of development. Here, we present DiffDomain, an algorithm leveraging high-dimensional random matrix theory to identify structurally reorganized TADs using high-throughput chromosome conformation capture (Hi-C) contact maps. Method comparison using multiple real Hi-C datasets reveals that DiffDomain outperforms alternative methods for false positive rates, true positive rates, and identifying a new subtype of reorganized TADs. Applying DiffDomain to Hi-C data from different cell types and disease states demonstrates its biological relevance. Identified reorganized TADs are associated with structural variations and epigenomic changes such as changes in CTCF binding sites. By applying to a single-cell Hi-C data from mouse neuronal development, DiffDomain can identify reorganized TADs between cell types with reasonable reproducibility using pseudo-bulk Hi-C data from as few as 100 cells per condition. Moreover, DiffDomain reveals differential cell-to-population variability and heterogeneous cell-to-cell variability in TADs. Therefore, DiffDomain is a statistically sound method for better comparative analysis of TADs using both Hi-C and single-cell Hi-C data.

Parallel synthetic aperture transport-of-intensity diffraction tomography with annular illumination.

Transport-of-intensity diffraction tomography (TIDT) is a recently developed label-free computational microscopy technique that retrieves high-resolution three-dimensional (3D) refractive index (RI) distribution of biological specimens from 3D intensity-only measurements. However, the non-interferometric synthetic aperture in TIDT is generally achieved sequentially through the acquisition of a large number of through-focus intensity stacks captured at different illumination angles, resulting in a very cumbersome and redundant data acquisition process. To this end, we present a parallel implementation of a synthetic aperture in TIDT (PSA-TIDT) with annular illumination. We found that the matched annular illumination provides a mirror-symmetric 3D optical transfer function, indicating the analyticity in the upper half-plane of the complex phase function, which allows for recovery of the 3D RI from a single intensity stack. We experimentally validated PSA-TIDT by conducting high-resolution tomographic imaging of various unlabeled biological samples, including human breast cancer cell lines (MCF-7), human hepatocyte carcinoma cell lines (HepG2), Henrietta Lacks (HeLa) cells, and red blood cells (RBCs).

Transport of intensity diffraction tomography with non-interferometric synthetic aperture for three-dimensional label-free microscopy.

We present a new label-free three-dimensional (3D) microscopy technique, termed transport of intensity diffraction tomography with non-interferometric synthetic aperture (TIDT-NSA). Without resorting to interferometric detection, TIDT-NSA retrieves the 3D refractive index (RI) distribution of biological specimens from 3D intensity-only measurements at various illumination angles, allowing incoherent-diffraction-limited quantitative 3D phase-contrast imaging. The unique combination of z-scanning the sample with illumination angle diversity in TIDT-NSA provides strong defocus phase contrast and better optical sectioning capabilities suitable for high-resolution tomography of thick biological samples. Based on an off-the-shelf bright-field microscope with a programmable light-emitting-diode (LED) illumination source, TIDT-NSA achieves an imaging resolution of 206 nm laterally and 520 nm axially with a high-NA oil immersion objective. We validate the 3D RI tomographic imaging performance on various unlabeled fixed and live samples, including human breast cancer cell lines MCF-7, human hepatocyte carcinoma cell lines HepG2, mouse macrophage cell lines RAW 264.7, Caenorhabditis elegans (C. elegans), and live Henrietta Lacks (HeLa) cells. These results establish TIDT-NSA as a new non-interferometric approach to optical diffraction tomography and 3D label-free microscopy, permitting quantitative characterization of cell morphology and time-dependent subcellular changes for widespread biological and medical applications.

Single-exposure 3D label-free microscopy based on color-multiplexed intensity diffraction tomography.

We present a 3D label-free refractive index (RI) imaging technique based on single-exposure intensity diffraction tomography (sIDT) using a color-multiplexed illumination scheme. In our method, the chromatic light-emitting diodes (LEDs) corresponding R/G/B channels in an annular programmable ring provide oblique illumination geometry that precisely matches the objective's numerical aperture. A color intensity image encoding the scattering field of the specimen from different directions is captured, and monochromatic intensity images concerning three color channels are separated and then used to recover the 3D RI distribution of the object following the process of IDT. In addition, the axial chromatic dispersion of focal lengths at different wavelengths introduced by the chromatic aberration of the objective lens and the spatial position misalignment of the ring LED source in the imaging system's transfer functions modeling are both corrected to significantly reduce the artifacts in the slice-based deconvolution procedure for the reconstruction of 3D RI distribution. Experimental results on MCF-7, Spirulina algae, and living Caenorhabditis elegans samples demonstrate the reliable performance of the sIDT method in label-free, high-throughput, and real-time (∼24 fps) 3D volumetric biological imaging applications.

Fully controlled photonic spin in highly confined optical field.

As an intrinsic attribute of light, the spin angular momentum (SAM) of photons has aroused considerable attention because of the fascinating properties emerging from light-matter interactions. We show that a diffraction-limited focal field with a steerable photonic spin structure in three dimensions can be produced under a 4π microscopic system. This is achieved by focusing two counter-propagating configurable vector beams produced in the coherent superposition of three different beams with x-polarization, y-polarization, and radial-polarization. By altering the amplitude factors of these resultant beams, the ratios between the three mutually orthogonal polarized components can be freely tuned within the focal plane, thereby allowing dynamic control over the spin orientation and ellipticity of the tightly focused optical field. The results demonstrated in this paper may find applications in spin-controlled nanophotonics.

Passively Q-switched Nd-doped fiber laser based on PbS/CdS core/shell quantum dots as a saturable absorber.

In our work, PbS/CdS core/shell quantum dots with an absorption peak of 1043 nm were successfully employed as a modulator for achieving a passively Q-switched Nd-doped fiber laser. The saturation intensity and modulation depth of the film-type modulator were 7.6 MW/cm2 and 4.1%, respectively. Due to the protection of the CdS shell, the PbS core exhibited good photo-chemical stability, which led to the generation of stable passively Q-switched operation. The maximum average output power was 7.88 mW with a minimum pulse width of 235.7 ns. To our knowledge, this was the first demonstration focusing on the combination of quantum dot and Nd-doped fiber laser; in addition, 235.7 ns was the narrowest pulse width of a passively Q-switched Nd-doped fiber laser. Our results highlighted the excellent nonlinear absorption properties of PbS/CdS core/shell quantum dots and give significant guidance for future optical applications of quantum dots and demonstrations of pulsed Nd-doped fiber lasers.

Redistributing the energy flow of a tightly focused radially polarized optical field by designing phase masks.

Redistributing the transverse energy flow in the focal plane of a tightly focused radially polarized optical field is described. We develop from theory a generalized analytical model for calculating the distributions of the electromagnetic field and the Poynting vector for a tightly focused radially polarized laser beam superposed with an optical vortex. We further explore the redistribution of the energy flow by designing phase masks, including traditional and annular vortex phase masks. Flexible control of the transverse energy flow rings is obtained with these phase masks. They provide a simple solution to transport absorptive particles along certain paths and therefore might be help in optical tweezer manipulations.

Focusing properties of arbitrary optical fields combining spiral phase and cylindrically symmetric state of polarization.

The tight focusing properties of optical fields combining a spiral phase and cylindrically symmetric state of polarization are presented. First, we theoretically analyze the mathematical characterization, Stokes parameters, and Poincaré sphere representations of arbitrary cylindrical vector (CV) vortex beams. Then, based on the vector diffraction theory, we derive and build an integrated analytical model to calculate the electromagnetic field and Poynting vector distributions of the input CV vortex beams. The calculations reveal that a generalized CV vortex beam can generate a sharper focal spot than that of a radially polarized (RP) plane beam in the focal plane. Besides, the focal size decrease accompanies its elongation along the optical axis. Hence, it seems that there is a trade-off between the transverse and axial resolutions. In addition, under the precondition that the absolute values between polarization order and topological charge are equal, a higher-order CV vortex can also achieve a smaller focal size than an RP plane beam. Further, the intensity for the sidelobe admits a significant suppression. To give a deep understanding of the peculiar focusing properties, the magnetic field and Poynting vector distributions are also demonstrated in detail. These properties may be helpful in applications such as optical trapping and manipulation of particles and superresolution microscopy imaging.

Optical cage generated by azimuthal- and radial-variant vector beams.

We propose a method to generate an optical cage using azimuthal- and radial-variant vector beams in a high numerical aperture optical system. A new kind of vector beam that has azimuthal- and radial-variant polarization states is proposed and demonstrated theoretically. Then, an integrated analytical model to calculate the electromagnetic field and Poynting vector distributions of the input azimuthal- and radial-variant vector beams is derived and built based on the vector diffraction theory of Richards and Wolf. From calculations, a full polarization-controlled optical cage is obtained by simply tailoring the radial index of the polarization, the uniformity U of which is up to 0.7748, and the cleanness C is zero. Additionally, a perfect optical cage can be achieved with U=1, and C=0 by introducing an amplitude modulation; its magnetic field and energy flow are also demonstrated in detail. Such optical cages may be helpful in applications such as optical trapping and high-resolution imaging.

Focus shaping by tailoring arbitrary hybrid polarization states that have a combination of orthogonal linear polarization bases.

We report a focus shaping method by tailoring hybrid states of polarization of arbitrary polarized beams that have a combination of orthogonal linear polarization bases. Such hybridly polarized beams comprising linear, elliptical, and circular polarizations in the beam cross section, have completely different optical properties compared to the scalar and locally linear-polarized vector beams. We demonstrate that, apart from the orientation of the local polarization state, another two degrees of freedom including the local ellipticity and the handedness in the beam cross section can be used in focus shaping. Square-shaped patterns, multiple foci, three-dimensional optical cages, optical needles, and channels can be obtained due to the increased control without any additional phase or amplitude modulations.

A new test of multivariate nonlinear causality.

The multivariate nonlinear Granger causality developed by Bai et al. (2010) (Mathematics and Computers in simulation. 2010; 81: 5-17) plays an important role in detecting the dynamic interrelationships between two groups of variables. Following the idea of Hiemstra-Jones (HJ) test proposed by Hiemstra and Jones (1994) (Journal of Finance. 1994; 49(5): 1639-1664), they attempt to establish a central limit theorem (CLT) of their test statistic by applying the asymptotical property of multivariate U-statistic. However, Bai et al. (2016) (2016; arXiv: 1701.03992) revisit the HJ test and find that the test statistic given by HJ is NOT a function of U-statistics which implies that the CLT neither proposed by Hiemstra and Jones (1994) nor the one extended by Bai et al. (2010) is valid for statistical inference. In this paper, we re-estimate the probabilities and reestablish the CLT of the new test statistic. Numerical simulation shows that our new estimates are consistent and our new test performs decent size and power.