Significance: Despite recent developments in microscopy, temporal aliasing can arise when imaging dynamic samples. Modern sampling frameworks, such as generalized sampling, mitigate aliasing but require measurement of temporally overlapping and potentially negative-valued inner products. Conventional cameras cannot collect these directly as they operate sequentially and are only sensitive to light intensity. Aim: We aim to mitigate aliasing in microscopy of dynamic monochrome samples by implementing generalized sampling via the use of a color camera and modulated color illumination. Approach: We solve the overlap problem by spectrally multiplexing the acquisitions and using (positive) B-spline segments as projection kernels. Reconstruction involves spectral unmixing and inverse filtering. We implemented this method using a color LED illuminator. We evaluated its performance by imaging a rotating grid and its applicability by imaging the beating zebrafish embryo heart in transmission and light-sheet microscopes. Results: Compared to stroboscopic imaging, our method mitigates aliasing with performance improving as the projection order increases. The approach can be implemented in conventional microscopes but is limited by the number of available LED colors and camera channels. Conclusions: Generalized sampling can be implemented via color modulation in microscopy to mitigate temporal aliasing. The simple hardware requirements could make it applicable to other optical imaging modalities.

中文翻译：

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通过使用时间调制彩色照明和标准 RGB 相机在动态生物样品的光学显微镜中减少混叠

意义：尽管最近在显微镜下取得了进展，但在对动态样本进行成像时可能会出现时间混叠。现代采样框架，例如广义采样，可以减轻混叠，但需要测量时间重叠和潜在的负值内积。传统相机无法直接收集这些，因为它们按顺序运行，并且仅对光强度敏感。目标：我们的目标是通过使用彩色相机和调制彩色照明实现广义采样来减轻动态单色样本显微镜中的混叠。方法：我们通过光谱复用采集并使用（正）B 样条段作为投影内核来解决重叠问题。重建涉及频谱解混和逆滤波。我们使用彩色 LED 照明器实现了这种方法。我们通过对旋转网格进行成像来评估其性能，并通过在透射和光片显微镜中对跳动的斑马鱼胚胎心脏进行成像来评估其适用性。结果：与频闪成像相比，我们的方法随着投影顺序的增加，性能提高，从而减轻了混叠。该方法可以在传统显微镜中实施，但受到可用 LED 颜色和相机通道数量的限制。结论：可以通过显微镜中的颜色调制实现广义采样，以减轻时间混叠。简单的硬件要求可以使其适用于其他光学成像模式。与频闪成像相比，我们的方法减轻了混叠，随着投影顺序的增加，性能也随之提高。该方法可以在传统显微镜中实施，但受到可用 LED 颜色和相机通道数量的限制。结论：可以通过显微镜中的颜色调制实现广义采样，以减轻时间混叠。简单的硬件要求可以使其适用于其他光学成像模式。与频闪成像相比，我们的方法减轻了混叠，随着投影顺序的增加，性能也随之提高。该方法可以在传统显微镜中实施，但受到可用 LED 颜色和相机通道数量的限制。结论：可以通过显微镜中的颜色调制实现广义采样，以减轻时间混叠。简单的硬件要求可以使其适用于其他光学成像模式。