A dual-modality fluorescence correlation microscope with single molecule sensitivity for the study of soft and biological systems

我们建议建立一个显微镜，允许以非常高的帧速率获取无背景荧光图像，同时仍然保持低噪声和高灵敏度。这种显微镜将集成两种不同的方法来抑制失焦背景，每种方法在不同的情况下都很有用：1）光片照明，以深入细胞或组织内成像，2）凸透镜诱导的软介质限制。该仪器将允许将单粒子和相关技术应用于软系统和生物系统的研究。具体应用将包括复杂介质中的异常扩散研究，以及早期苍蝇胚胎中形态发生素的时空运输和活动。通过推动目前在这些系统中可以观察到的动力学极限，我们将获得尚未观察到的现象。建立一个国家的最先进的仪器的过程将使学员在我的小组有机会获得第一手和深入的知识，在生物光子学，促进他们的就业前景。

We propose to build a microscope allowing to acquire background-free fluorescence images at a very high frame rate, while still maintaining low noise and high sensitivity. This microscope will integrate two different methods for suppressing out-of-focus background, each useful in different situations: 1) lightsheet illumination to image deep within cells or tissues and 2) convex lens-induced confinement for soft media. This instrument will allow applying single particle and correlation techniques to the study of soft and biological systems. Specific applications will include the study of anomalous diffusion in complex media, and the spatio-temporal trafficking and activity of morphogens in the early fly embryo. By pushing the limit of what is currently possible to observe in these systems in terms of dynamics, we will gain access to yet unobserved phenomena. The process of building a state-of-the-art instrument will give trainees in my group the opportunity to gain a first-hand and in-depth knowledge in biophotonics, furthering their employment perspectives.