Dual infrared visible confocal microscopy for imaging inter-cellular communication

细胞间信号传导对于理解生物过程的分子基础至关重要 营养吸收、植物和动物对刺激的反应、疾病发生、神经生物学和认知， 免疫反应和细胞通讯。我们的 MIRA R35 专注于开发纳米级近红外 我们已经开发了可以报告细胞外化学通讯的生物传感器 在我们获奖的前 3 年里，我们使用了许多近红外纳米传感器来对细胞外化学分析物进行成像。 2019 年底，新的商用相机硬件上市，首次实现了并发 使用同一探测器对可见光和近红外波长进行成像。我们正在请求一个设备 补充以构建多光子显微镜以实现并发近红外荧光 使用我们的纳米传感器对细胞流出进行成像，并具有细胞转录成像的新功能 以及伴随细胞因子信号传导的可见荧光成像的形态变化。在前 3 获奖多年以来，我们开发了许多用于细胞外化学分析物的近红外纳米传感器 包括多巴胺、神经肽催产素、细胞因子 VEGF、葡萄糖和过氧化氢。目的 我们的设备补充要求是利用新的荧光成像技术，特别是 可见光和近红外敏感相机，这将使我们能够对细胞内和细胞外进行成像 实时细胞信号传导的变化。分泌细胞因子的直接细胞成像（之前的 R35 目标） 以及单细胞转录和形态学的变化（新的 R35 目标及技术补充） 将为如何刺激单个或少数单个细胞的细胞因子分泌谱提供新的范例 由趋化因子和细胞因子组成，构成目前使用的细胞因子分泌谱的基础 基于生物标记的诊断。 兰德里摘要 AB-1

Intercellular signaling is central to understanding the molecular basis of biological processes ranging from nutrient uptake, response to stimulus in plants and animals, disease inception, neurobiology and cognition, immune responses, and cellular communication. Our MIRA R35 centers on developing nm-scale near-infrared biosensors that can report on extracellular chemical communication, through which we have already developed numerous near-infrared nanosensors to image extracellular chemical analytes in the first 3 years of our award. In late 2019, new commercial camera hardware became available that – for the first time – enables concurrent imaging of visible and near-infrared wavelengths with the same detector. We are requesting an equipment supplement to enable building a multiphoton microscope to enable concurrent near-infrared fluorescence imaging of cellular efflux with our nanosensors together with a new capability in imaging cellular transcriptional and morphological changes with visible fluorescence imaging that accompany cytokine signaling. In the first 3 years of our award, we have developed numerous near-infrared nanosensors for extracellular chemical analytes including dopamine, neuropeptide oxytocin, cytokine VEGF, glucose, and hydrogen peroxide. The purpose of our equipment supplement request is to take advantage of new fluorescence imaging technology, specifically a visible and near-infrared sensitive camera, that will enable us to image both intracellular and extracellular changes that underlie cell signaling in real-time. Direct cellular imaging of secreted cytokines (prior R35 goal) together with single-cell changes in transcription and morphology (new R35 goal with technology supplement) will provide a new paradigm on how cytokine secretion profiles from single or few individual cells are stimulated by chemokines and cytokines, which forms the basis of the cytokine secretion profiles currently used in biomarker-based diagnostics. Landry ABSTRACT AB-1