TomoFlow: Tomographic Imaging Two-Photon Flow Cytometry

点击翻译按钮获取中文摘要

Multiphoton microscopy allows deep optical penetration making it the method of choice for optical imaging in tissue. In order to understand the molecular and cellular origin of disease, it is crucial to sense and visualize sub-cellular units and cell-to-cell interactions at high temporal and spatial resolution and at relevant scales. In addition, for a robust and reliable statistic it is necessary to study a large, heterogenous population of cells. This can be optimally addressed by imaging flow cytometry (IFC), employing microscopic imaging of cells in microfluidic flow. Here, we propose a high throughput IFC technique with three-dimensional (3D) acquisition for the study of cell populations and organoids by employing high speed Two-Photon microscopy and volumetric IFC. The high penetration of Two-Photon microscopy and the volumetric acquisition lead to a tomographic imaging acquisition. Thus, we coin this new technology platform TomoFlow for tomographic flow imaging. The high-throughput is enabled by the recently introduced spectro-temporal laser imaging by diffractive excitation (SLIDE) technique providing imaging rates of 4.000 frames per second. For TomoFlow, we will develop new microfluidic flow chips to enable tomographic imaging at high three-dimensional resolution (< 1µm3). We will further apply the TomoFlow technology to the detection of tumor cells in flow, organoid and tissue fragment imaging suspended in whole, unfiltered blood as well as pharmacological studies on organoids with high-resolution at the cellular and sub-cellular level and high throughput for rich statistics. This research endeavor has the potential of providing a new biomedical diagnostic tool for the study of the cellular origin of disease and the development of targeted, personalized therapies and medicine on the cellular and molecular level.