Ultrashort pulse, multispectral non-linear optical microscopy (NLOM) is developed and used to image, simultaneously, a mixed population of cells expressing different fluorescent protein mutants in a 3D tissue model of angiogenesis. Broadband, sub-10-fs pulses are used to excite multiple fluorescent proteins and generate second harmonic in collagen. A 16-channel multispectral detector is used to delineate the multiple non-linear optical signals, pixel by pixel, in NLOM. The ability to image multiple fluorescent protein mutants and collagen, enables serial measurements of cell-cell and cell-matrix interactions in our 3D tissue model and characterization of fundamental processes in angiogenic morphogenesis.

中文翻译：

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超短脉冲多光谱非线性光学显微镜

超短脉冲、多光谱非线性光学显微镜 (NLOM) 被开发并用于同时对血管生成的 3D 组织模型中表达不同荧光蛋白突变体的混合细胞群进行成像。宽带、亚 10-fs 脉冲用于激发多种荧光蛋白并在胶原蛋白中产生二次谐波。一个 16 通道的多光谱检测器用于在 NLOM 中逐个像素地描绘多个非线性光信号。能够对多种荧光蛋白突变体和胶原蛋白进行成像，可以在我们的 3D 组织模型中连续测量细胞-细胞和细胞-基质相互作用，并表征血管生成形态发生的基本过程。