Dual infrared visible confocal microscopy for imaging inter-cellular communication

细胞间信号转导是理解生物过程的分子基础的核心，包括 营养吸收，植物和动物对刺激的反应，疾病开始，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 可以报告细胞外化学交流的生物传感器，我们已经通过这种传感器开发出 在我们获奖的前3年，我们使用了大量近红外纳米传感器对细胞外化学分析物进行成像。 2019年末，新的商用摄像机硬件首次面市，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。我们需要一台设备 能够建立多光子显微镜以实现同时的近红外荧光的补充 利用我们的纳米传感器对细胞外排进行成像，以及对细胞转录进行成像的新能力 以及伴随细胞因子信号的可见荧光成像的形态变化。在前3个月 多年来，我们已经开发了许多用于细胞外化学分析的近红外纳米传感器 包括多巴胺、神经肽催产素、细胞因子血管内皮生长因子、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 可见光和近红外感光摄像头，这将使我们能够同时成像细胞内和细胞外 实时改变细胞信号的基础。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的变化(新的R35目标和技术补充) 将为如何刺激单个或少数单个细胞的细胞因子分泌提供一个新的范例 通过趋化因子和细胞因子，这构成了目前在 基于生物标记物的诊断。 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