Quantitative Light Imaging Laboratory, Department of Electrical and Computer Engineering, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Department of Bioengineering, Micro and Nanotechnology Lab, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Sci Rep. 2017 Mar 24;7:44034. doi: 10.1038/srep44034.
We present a new approach for retrieving halo-free phase contrast microscopy (hfPC) images by upgrading the conventional PC microscope with an external interferometric module, which generates sufficient data for reversing the halo artifact. Acquiring four independent intensity images, our approach first measures haloed phase maps of the sample. We solve for the halo-free sample transmission function by using a physical model of the image formation under partial spatial coherence. Using this halo-free sample transmission, we can numerically generate artifact-free PC images. Furthermore, this transmission can be further used to obtain quantitative information about the sample, e.g., the thickness with known refractive indices, dry mass of live cells during their cycles. We tested our hfPC method on various control samples, e.g., beads, pillars and validated its potential for biological investigation by imaging live HeLa cells, red blood cells, and neurons.
我们提出了一种新的方法来获取无晕 halo-free 相衬显微镜（hfPC）图像，通过在传统的 PC 显微镜上升级一个外部干涉模块，该模块可以生成足够的数据来反转晕环artifact。通过获取四个独立的强度图像，我们的方法首先测量样品的晕环相位图。我们使用部分空间相干下的图像形成的物理模型来求解无晕环样品的传输函数。使用这个无晕环样品的传输函数，我们可以在数字上生成无晕环artifact 的 PC 图像。此外，这个传输函数还可以进一步用于获取关于样品的定量信息，例如，已知折射率的厚度，细胞周期中活细胞的干燥质量。我们在各种对照样品上测试了我们的 hfPC 方法，例如珠子、柱子，并通过对活 HeLa 细胞、红细胞和神经元进行成像来验证其用于生物学研究的潜力。
