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学术报告:在体高时空分辨荧光成像

发布时间:2021-05-20    点击数:

报告人:陈良怡北京大学

邀请人:屈军乐教授

时 间:2021年5月23日上午9:30

地 点:粤海校区光电所多功能厅

报告摘要:

Here we will present three pieces of high-resolution fluorescence microscopy methods we invented for live sample imaging.The first one is for in vivo imaging, which is a series of fast, high-resolution, miniaturized two-photon microscope (FHIRM-TPM), which can be used to resolve single spine in freely-behaving animals and achieved volumetric or mulitplane imaging over an axial distance of 180 μm with an interplane switch time of less than 1.5 ms. Further, we engineered the headpiece for repeated mounting and dismounting, and demonstrated its robustness by recording neuronal activities from the same brain region over a time frame of several weeks.

The second method is for live cell long-term super-resolution (SR) imaging. We have developed a deconvolution algorithm for structured illumination microscopy based on Hessian matrixes (Hessian-SIM). It uses the continuity of biological structures in multiple dimensions as a priori knowledge to guide image reconstruction and attains artifact-minimized SR images with less than 10% of the photon dose used by conventional SIM while substantially outperforming current algorithms at low signal intensities. Its high sensitivity allows the use of sub-millisecond excitation pulses followed by dark recovery times to reduce photobleaching of fluorescent proteins, enabling hour-long time-lapse SR imaging in live cells.

The third technology is a dual-mode SR microscopy for highlighting molecules as well as a holistic view of related interacting organelles in live cells. It is a combination of two-dimensional Hessian-SIM with label-free three-dimensional optical diffraction tomography (ODT), term SR fluorescence-assisted diffraction computational tomography (SR-FACT). The ODT module is capable of resolving mitochondria, lipid droplets, the nuclear membrane, chromosomes, the tubular endoplasmic reticulum and lysosomes. These works demonstrate the unique capabilities of SR-FACT, which suggest its wide applicability in cell biology in general.

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