.
Significance:
Differential interference contrast (DIC) microscopes allow noninvasive
in vivo
observation of transparent microstructures in tissue without the use of fluorescent dyes or genetic modification. We show how to modify a DIC microscope to measure the sample phase distribution accurately and in real-time even deep inside sample tissue.
Aim:
Our aim is to improve the DIC microscope’s phase measurement to remove the phase bias that occurs in the presence of strong scattering.
Approach:
A quarter-wave plate was added in front of the polarization camera, allowing a modified phase calculation to incorporate all four polarization orientation angles (0 deg, 45 deg, 90 deg, and 135 deg) captured simultaneously by the polarization camera, followed by deconvolution.
Results:
We confirm that the proposed method reduces phase measurement error in the presence of scattering and demonstrate the method using
in vivo
imaging of a beating heart inside a medaka egg and the whole-body blood circulation in a young medaka fish.
Conclusions:
Modifying a polarization-camera DIC microscope with a quarter-wave plate allows users to image deep inside samples without phase bias due to scattering effects.