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

我们建议建造一种显微镜，允许以非常高的帧速率获取无背景的荧光图像，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有用：1)光片照明，以成像细胞或组织内部的深处；2)凸透镜诱导的软性介质的限制。该仪器将允许将单粒子和关联技术应用于软系统和生物系统的研究。具体的应用将包括研究复杂介质中的异常扩散，以及早期苍蝇胚胎中形态原的时空迁移和活性。通过在动力学方面突破目前在这些系统中可以观察到的极限，我们将获得尚未观察到的现象。建造最先进的仪器的过程将使我团队中的实习生有机会获得生物光子学方面的第一手和深入的知识，进一步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.