Axially resolved volumetric two-photon microscopy with an extended field of view using depth localization under mirrored Airy beams.

It is a great challenge in two-photon microscopy (2PM) to have a high volumetric imaging speed without sacrificing the spatial and temporal resolution in three dimensions (3D). The structure in 2PM images could be reconstructed with better spatial and temporal resolution by the proper choice of the data processing algorithm. Here, we propose a method to reconstruct 3D volume from 2D projections imaged by mirrored Airy beams. We verified that our approach can achieve high accuracy in 3D localization over a large axial range and is applicable to continuous and dense sample. The effective field of view after reconstruction is expanded. It is a promising technique for rapid volumetric 2PM with axial localization at high resolution.

Resolution enhancement in an extended depth of field for volumetric two-photon microscopy.

The resolution enhancement over the extended depth of field (DOF) in the volumetric two-photon microscopy (TPM) is demonstrated by utilizing multiple orders of Bessel beams. Here the conventional method of switching laser modes (SLAM) in 2D is introduced to 3D, denoted as the volumetric SLAM (V-SLAM). The equivalent scanning beam in the TPM is a thin needle-like beam, which is generated from the subtraction between the needle-like 0th-order and the straw-like 1st-order Bessel beams. Compared with the 0th-order Bessel beam, the lateral resolution of the V-SLAM is increased by 28.6% and maintains over the axial depth of 56 µm. The V-SLAM performance is evaluated by employing fluorescent beads and a mouse brain slice. The V-SLAM approach provides a promising solution to improve the lateral resolutions for fast volumetric imaging on sparsely distributed samples.

Volumetric two-photon microscopy with a non-diffracting Airy beam.

We demonstrate a volumetric two-photon microscopy (TPM) using the non-diffracting Airy beam as illumination. Direct mapping of the imaging trajectory shows that the Airy beam extends the axial imaging range around six times longer than a traditional Gaussian beam does along the propagation direction, while maintaining a comparable lateral width. Benefiting from its non-diffracting nature, the TPM with Airy beam illumination is able not only to capture a volumetric image within a single frame, but also to acquire image structures behind a strongly scattered medium. The volumetric specimen is mapped layer by layer under Gaussian mode, while the three-dimensional structure is projected to a single two-dimensional image under Airy mode, leading to a significantly increased acquisition speed. The performance of the TPM is evaluated employing a phantom of agarose gel imbedding fluorescent beads as well as a mouse brain slice. Finally, we showcase the penetration ability of the developed Airy TPM by imaging through a scattering environment.

The pattern of cortical dysfunction in a mouse model of a schizophrenia-related microdeletion.

We used a mouse model of the schizophrenia-predisposing 22q11.2 microdeletion to evaluate how this genetic lesion affects cortical neural circuits at the synaptic, cellular, and molecular levels. Guided by cognitive deficits, we demonstrated that mutant mice display robust deficits in high-frequency synaptic transmission and short-term plasticity (synaptic depression and potentiation), as well as alterations in long-term plasticity and dendritic spine stability. Apart from previously reported reduction in dendritic complexity of layer 5 pyramidal neurons, altered synaptic plasticity occurs in the context of relatively circumscribed and often subtle cytoarchitectural changes in neuronal density and inhibitory neuron numbers. We confirmed the pronounced DiGeorge critical region 8 (Dgcr8)-dependent deficits in primary micro-RNA processing and identified additional changes in gene expression and RNA splicing that may underlie the effects of this mutation. Reduction in Dgcr8 levels appears to be a major driver of altered short-term synaptic plasticity in prefrontal cortex and working memory but not of long-term plasticity and cytoarchitecture. Our findings inform the cortical synaptic and neuronal mechanisms of working memory impairment in the context of psychiatric disorders. They also provide insight into the link between micro-RNA dysregulation and genetic liability to schizophrenia and cognitive dysfunction.

The effect of transcranial direct current and magnetic stimulation on fear extinction and return of fear: A meta-analysis and systematic review.

BACKGROUND: We conducted a meta-analysis and qualitative review on the randomized controlled trials investigating the effects of transcranial direct current stimulation and transcranial magnetic stimulation on fear extinction and the return of fear in non-primate animals and humans. METHODS: The meta-analysis was conducted by searching PubMed, Web of science, PsycINFO, and Cochrane Library and extracting fear response in the active and sham groups in the randomized controlled trials. The pooled effect size was quantified by Hedges' g using a three-level meta-analytic model in R. RESULTS: We identified 18 articles on the tDCS effect and 5 articles on the TMS effect, with 466 animal subjects and 621 human subjects. Our findings show that tDCS of the prefrontal cortex significantly inhibit fear retrieval in animal models (Hedges' g = -0.50). In human studies, TMS targeting the dorsolateral/ventromedial prefrontal cortex has an inhibiting effect on the return of fear (Hedges' g = -0.24). LIMITATIONS: The limited number of studies and the heterogeneous designs of the selected studies made cross-study and cross-species comparison difficult. CONCLUSIONS: Our findings shed light on the optimal non-invasive brain stimulation protocols for targeting the neural circuitry of threat extinction in humans.

High-contrast, fast chemical imaging by coherent Raman scattering using a self-synchronized two-colour fibre laser.

Coherent Raman scattering (CRS) microscopy is widely recognized as a powerful tool for tackling biomedical problems based on its chemically specific label-free contrast, high spatial and spectral resolution, and high sensitivity. However, the clinical translation of CRS imaging technologies has long been hindered by traditional solid-state lasers with environmentally sensitive operations and large footprints. Ultrafast fibre lasers can potentially overcome these shortcomings but have not yet been fully exploited for CRS imaging, as previous implementations have suffered from high intensity noise, a narrow tuning range and low power, resulting in low image qualities and slow imaging speeds. Here, we present a novel high-power self-synchronized two-colour pulsed fibre laser that achieves excellent performance in terms of intensity stability (improved by 50 dB), timing jitter (24.3 fs), average power fluctuation (<0.5%), modulation depth (>20 dB) and pulse width variation (<1.8%) over an extended wavenumber range (2700-3550 cm-1). The versatility of the laser source enables, for the first time, high-contrast, fast CRS imaging without complicated noise reduction via balanced detection schemes. These capabilities are demonstrated in this work by imaging a wide range of species such as living human cells and mouse arterial tissues and performing multimodal nonlinear imaging of mouse tail, kidney and brain tissue sections by utilizing second-harmonic generation and two-photon excited fluorescence, which provides multiple optical contrast mechanisms simultaneously and maximizes the gathered information content for biological visualization and medical diagnosis. This work also establishes a general scenario for remodelling existing lasers into synchronized two-colour lasers and thus promotes a wider popularization and application of CRS imaging technologies.

Compact fs ytterbium fiber laser at 1010 nm for biomedical applications.

Ytterbium-doped fiber lasers (YDFLs) working in the near-infrared (NIR) spectral window and capable of high-power operation are popular in recent years. They have been broadly used in a variety of scientific and industrial research areas, including light bullet generation, optical frequency comb formation, materials fabrication, free-space laser communication, and biomedical diagnostics as well. The growing interest in YDFLs has also been cultivated for the generation of high-power femtosecond (fs) pulses. Unfortunately, the operating wavelengths of fs YDFLs have mostly been confined to two spectral bands, i.e., 970-980 nm through the three-level energy transition and 1030-1100 nm through the quasi three-level energy transition, leading to a spectral gap (990-1020 nm) in between, which is attributed to an intrinsically weak gain in this wavelength range. Here we demonstrate a high-power mode-locked fs YDFL operating at 1010 nm, which is accomplished in a compact and cost-effective package. It exhibits superior performance in terms of both short-term and long-term stability, i.e., <0.3% (peak intensity over 2.4 µs) and <4.0% (average power over 24 hours), respectively. To illustrate the practical applications, it is subsequently employed as a versatile fs laser for high-quality nonlinear imaging of biological samples, including two-photon excited fluorescence microscopy of mouse kidney and brain sections, as well as polarization-sensitive second-harmonic generation microscopy of potato starch granules and mouse tail muscle. It is anticipated that these efforts will largely extend the capability of fs YDFLs which is continuously tunable over 970-1100 nm wavelength range for wideband hyperspectral operations, serving as a promising complement to the gold-standard Ti:sapphire fs lasers.