Stephenson School of Biomedical Engineering, University of Oklahoma, Norman, Oklahoma, USA.
School of Electrical and Computer Engineering, University of Oklahoma, Norman, Oklahoma, USA.
J Biophotonics. 2023 May;16(5):e202200303. doi: 10.1002/jbio.202200303. Epub 2023 Jan 10.
This study aims to develop a high throughput Fourier ptychographic microscopy (FPM) technique based on symmetric illumination and a color detector, which is able to accelerate image acquisition by up to 12 times. As an emerging technology, the efficiency of FPM is limited by its data acquisition process, especially for color microscope image reconstruction. To overcome this, we built an FPM prototype equipped with a color camera and a 4×/0.13 NA objective lens. During the image acquisition, two symmetric LEDs illuminate the sample simultaneously using white light, which doubles the light intensity and reduces the total captured raw patterns by half. A standard USAF 1951 resolution target was used to measure the system's modulation transfer function (MTF) curve, and the H&E-stained ovarian cancer samples were then imaged to assess the feature qualities depicted on the reconstructed images. The results showed that the measured MTF curves of red, green, and blue channels are generally comparable to the corresponding curves generated by conventional FPM, while symmetric illumination FPM preserves more tissue details, which is superior to the results captured by conventional 20×/0.4 NA objective lens. This investigation initially verified the feasibility of symmetric illumination based color FPM.
本研究旨在开发一种基于对称照明和彩色探测器的高通量傅里叶叠层显微镜（FPM）技术，该技术能够将图像采集速度提高 12 倍。作为一种新兴技术，FPM 的效率受到其数据采集过程的限制，特别是对于彩色显微镜图像重建。为了克服这一问题，我们构建了一个配备彩色相机和 4×/0.13 NA 物镜的 FPM 原型。在图像采集过程中，两个对称的 LED 同时用白光照射样本，从而将光强度提高一倍，并将总采集的原始图案减少一半。我们使用标准的 USAF 1951 分辨率测试标靶来测量系统的调制传递函数（MTF）曲线，然后对 H&E 染色的卵巢癌细胞样本进行成像，以评估重建图像上所描绘的特征质量。结果表明，红色、绿色和蓝色通道的测量 MTF 曲线通常与传统 FPM 生成的相应曲线相当，而对称照明 FPM 保留了更多的组织细节，优于传统 20×/0.4 NA 物镜捕获的结果。本研究初步验证了基于对称照明的彩色 FPM 的可行性。
