Opt Lett. 2022 May 15;47(10):2514-2517. doi: 10.1364/OL.456705.
Multi-color fluorescence microscopy presents highly detailed biological samples interactively. However, current multi-color methods suffer from an intricate optical setup, complicated image analysis, or a long acquisition time. To address these issues, here we develop a simple multi-color method based on a customized colorimetry camera to enable the detection of multiple structures from single-shot acquisition. The unfiltered channel (W pixels) and color channels (R, G, B, and NIR pixels) in this customized camera simultaneously provide a broad detection wavelength range and high detection sensitivity. We built a simple optical setup by replacing the monochrome camera in a basic fluorescence microscopy system with a colorimetry camera, and developed effective image analysis procedures to reconstruct a multi-color image from a single frame of a raw image. We demonstrated single-shot four-color wide-field fluorescence imaging on fixed cos-7 cells with < 5% cross talk, which is comparable to the best reported values. Our method greatly simplifies both the optical system and image analysis in the widely used method of multi-color fluorescence microscopy, thus offering an effective and easy way to study multiple objects at the same time.
多色荧光显微镜呈现出高度详细的生物样本，具有交互性。然而，目前的多色方法存在复杂的光学设置、复杂的图像分析或较长的采集时间等问题。为了解决这些问题，我们开发了一种基于定制比色相机的简单多色方法，能够从单次采集检测多个结构。该定制相机中的未滤光通道（W 像素）和彩色通道（R、G、B 和近红外像素）同时提供了广泛的检测波长范围和高检测灵敏度。我们通过用比色相机替换基本荧光显微镜系统中的单色相机，构建了一个简单的光学设置，并开发了有效的图像分析程序，从原始图像的单个帧重建多色图像。我们在固定的 cos-7 细胞上演示了单次四色宽场荧光成像，串扰小于 5%，与报道的最佳值相当。我们的方法大大简化了多色荧光显微镜广泛使用的方法中的光学系统和图像分析，从而为同时研究多个物体提供了一种有效且简单的方法。
