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首页> 外文会议>Conference on multiphoton microscopy in the biomedical sciences XVI >In vivo imaging flow cytometry based on laser scanning two-photon microscopy at kHz cross-sectional frame rate
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In vivo imaging flow cytometry based on laser scanning two-photon microscopy at kHz cross-sectional frame rate

机译：基于激光扫描的kHz横截面帧速率的激光扫描二光子显微镜的体内成像流式细胞仪

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摘要

In vivo flow cytometry has found numerous applications in biology and pharmacology. However, conventional cytometry does not provide the detailed morphological information that is needed to fully determine the phenotype of individual circulating cells. Imaging cytometry, capable of visualizing the morphology and dynamics of the circulating cells at high spatiotemporal resolution, is highly desired. Current wide-field based image cytometers are limited in the imaging depth and provide only two-dimensional resolution. For deep tissue imaging, laser scanning two-photon fluorescence microscopy (TPM) is widely adopted. However, for applications in flow cytometry, the axial scanning speed of current TPMs is inadequate to provide high-speed cross-sectional imaging of vasculature. We have integrated an optical phase-locked ultrasound lens into a standard TPM and achieved microsecond-scale axial scanning. With a galvo scanner for transverse scanning, we achieved kHz cross-sectional frame rate. Here we report its applications for in vivo deformability cytometry and in vivo imaging flow cytometry, and demonstrate the capability of imaging dynamical morphologies of flowing cells, distinguishing cells and cellular clusters, and simultaneously quantifying different cell populations based on their fluorescent labels.
机译：体内流式细胞仪在生物学和药理学中发现了许多应用。然而，常规的细胞学不提供完全确定单个循环细胞表型所需的详细的形态学信息。非常需要成像细胞术，能够以高时的血流分辨率可视化循环细胞的形态和动力学。基于宽场的图像细胞计是有限的成像深度，并且仅提供二维分辨率。对于深组织成像，广泛采用激光扫描双光子荧光显微镜（TPM）。然而，对于流式细胞术中的应用，电流TPM的轴向扫描速度不足以提供脉管系统的高速横截面成像。我们已经将光学锁相超声镜片集成为标准TPM并实现了微秒级轴向扫描。使用Galvo扫描仪进行横向扫描，我们实现了KHz横截面帧速率。在这里，我们将其用于体内可变形性细胞术和体内成像流式细胞术中的应用，并证明了流动细胞的成像动态形态，区分细胞和细胞簇的能力，并同时量化了基于其荧光标记的不同细胞群。

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