Fiber photometry has become increasingly popular among neuroscientists as a convenient tool for the recording of genetically defined neuronal population in behaving animals. Here, we report the development of the multi-channel fiber photometry system to simultaneously monitor neural activities in several brain areas of an animal or in different animals. In this system, a galvano-mirror modulates and cyclically couples the excitation light to individual multimode optical fiber bundles. A single photodetector collects excited light and the configuration of fiber bundle assembly and the scanner determines the total channel number. We demonstrated that the system exhibited negligible crosstalk between channels and optical signals could be sampled simultaneously with a sample rate of at least 100 Hz for each channel, which is sufficient for recording calcium signals. Using this system, we successfully recorded GCaMP6 fluorescent signals from the bilateral barrel cortices of a head-restrained mouse in a dual-channel mode, and the orbitofrontal cortices of multiple freely moving mice in a triple-channel mode. The multi-channel fiber photometry system would be a valuable tool for simultaneous recordings of population activities in different brain areas of a given animal and different interacting individuals.

In vivo optical spatio-temporal imaging of the tumor microenvironment is useful to explain how tumor immunotherapies work. However, the lack of fluorescent antigens with strong immunogenicity makes it difficult to study the dynamics of how tumors are eliminated by any given immune response. Here, we develop an effective fluorescent model antigen based on the tetrameric far-red fluorescent protein KatushkaS158A (tfRFP), which elicits both humoral and cellular immunity. We use this fluorescent antigen to visualize the dynamic behavior of immunocytes as they attack and selectively eliminate tfRFP-expressing tumors in vivo; swarms of immunocytes rush toward tumors with high motility, clusters of immunocytes form quickly, and numerous antigen-antibody complexes in the form of tfRFP⁺ microparticles are generated in the tumor areas and ingested by macrophages in the tumor microenvironment. Therefore, tfRFP, as both a model antigen and fluorescent reporter, is a useful tool to visualize specific immune responses in vivo.

The ability to remember and to navigate to safe places is necessary for survival. Place navigation is known to involve medial entorhinal cortex (MEC)-hippocampal connections. However, learning-dependent changes in neuronal activity in the distinct circuits remain unknown. Here, by using optic fiber photometry in freely behaving mice, we discovered the experience-dependent induction of a persistent-task-associated (PTA) activity. This PTA activity critically depends on learned visual cues and builds up selectively in the MEC layer II-dentate gyrus, but not in the MEC layer III-CA1 pathway, and its optogenetic suppression disrupts navigation to the target location. The findings suggest that the visual system, the MEC layer II, and the dentate gyrus are essential hubs of a memory circuit for visually guided navigation.

We extend classical light ghost imaging to the area of fluorescence imaging and propose a new fluorescence imaging method. For the first time, we demonstrate both theoretically and experimentally that fluorescence ghost imaging can be realized with pseudothermal light. Important factors influencing the visibility and resolution of the images are discussed to improve the quality of the fluorescence ghost imaging. We hope that this work may pave the road for ghost imaging to biomedical applications.

We extend the subtractive imaging method to label-free second harmonic generation (SHG) microscopy to enhance the spatial resolution and contrast. This method is based on the intensity difference between two images obtained with circularly polarized Gaussian and doughnut-shaped beams, respectively. By characterizing the intensity and polarization distributions of the two focused beams, we verify the feasibility of the subtractive imaging method in polarization dependent SHG microscopy. The resolution and contrast enhancement in different biological samples is demonstrated. This work will open a new avenue for the applications of SHG microscopy in biomedical research.

Endoscopic imaging is the main method for detecting gastrointestinal diseases, which adversely affect human health. White light endoscopy (WLE) was the first method used for endoscopic examination and is still the preliminary step in the detection of gastrointestinal diseases during clinical examination. However, it cannot accurately diagnose gastrointestinal diseases owing to its poor correlation with histopathological diagnosis. In recent years, many advanced endoscopic methods have emerged to improve the detection accuracy by endoscopy. Chromoendoscopy (CE) enhances the contrast between normal and diseased tissues using biocompatible dye agents. Narrow band imaging (NBI) can improve the contrast between capillaries and submucosal vessels by changing the light source acting on the tissue using special filters to realize the visualization of the vascular structure. Flexible spectral imaging color enhancement (FICE) technique uses the reflectance spectrum estimation technique to obtain individual spectral images and reconstructs an enhanced image of the mucosal surface using three selected spectral images. The i-Scan technology takes advantage of the different reflective properties of normal and diseased tissues to obtain images, and enhances image contrast through post-processing algorithms. These abovementioned methods can be used to detect gastrointestinal diseases by observing the macroscopic structure of the digestive tract mucosa, but the ability of early cancer detection is limited with low resolution. However, based on the principle of confocal imaging, probe-based confocal laser endomicroscopy (pCLE) can enable cellular visualization with high-performance probes, which can present cellular morphology that is highly consistent with that shown by biopsy to provide the possibility of early detection of cancer. Other endoscopic imaging techniques including endoscopic optical coherence tomography (EOCT) and photoacoustic endoscopy (PAE), are also promising for diagnosing gastrointestinal diseases. This review focuses on these technologies and aims to provide an overview of different technologies and their clinical applicability.