Halo-free Phase Contrast Microscopy.
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.
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.
Tan H
TH
Nguyen
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.
Mikhail
M
Kandel
Department of Bioengineering, Micro and Nanotechnology Lab, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Haadi M
HM
Shakir
Department of Bioengineering, Micro and Nanotechnology Lab, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Catherine
C
Best-Popescu
Munroe-Meyer Institute, University of Nebraska Medical Center (UNMC), Omaha, Nebraska 68198, USA.
Jyothi
J
Arikkath
Computational Imaging Group, Department of Electrical and Computer Engineering, Coordinated Science Lab, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Minh N
MN
Do
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.
Gabriel
G
Popescu
D016477:Artifacts; D004912:Erythrocytes / Q000166:cytology; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008858:Microscopy, Phase-Contrast / Q000295:instrumentation / Q000379:methods*; D008962:Models, Theoretical; D009474:Neurons / Q000166:cytology; D012031:Refractometry