Abstract
We present a quantitative phase-contrast confocal microscope (QPCCM) by combining a line-scanning confocal system with digital holography (DH). This combination can merge the merits of these two different imaging modalities. High-contrast intensity images with low coherent noise, and the optical sectioning capability are made available due to the confocality. Phase profiles of the samples become accessible thanks to DH. QPCCM is able to quantitatively measure the phase variations of optical sections of the opaque samples and has the potential to take high-quality intensity and phase images of non-opaque samples such as many biological samples. Because each line scan is recorded by a hologram that may contain the optical aberrations of the system, it opens avenues for a variety of numerical aberration compensation methods and development of full digital adaptive optics confocal system to emulate current hardware-based adaptive optics system for biomedical imaging, especially ophthalmic imaging. Preliminary experiments with a microscope objective of NA 0.65 and 40 × on opaque samples are presented to demonstrate this idea. The measured lateral and axial resolutions of the intensity images from the current system are ~0.64μm and ~2.70μm respectively. The noise level of the phase profile by QPCCM is ~2.4nm which is better than the result by DH.
Highlights
Point-scanning confocal microscopy was originated by M
To demonstrate the basic process of the confocal image reconstructions, a negative 1951 United States Air Force (USAF) resolution target is used as the sample
In subsection 3.1, we have demonstrated that the intensity image of quantitative phase-contrast confocal microscope (QPCCM) is better than the wide-field coherent image in terms of the contrast and coherent noise
Summary
Point-scanning confocal microscopy was originated by M. To speed up image acquisition and simplify the optical system, line-scanning confocal systems have been proposed and tested in industrial inspection, imaging of human tissues, and ophthalmology [6,7,8,9]. Similar to the point-scanning confocal system, the linescanning confocal system is unable to get the quantitative phase information of the optical field that is of great interest in industrial inspection and biomedical imaging. The first effort of combining the confocality with off-axis DH was made in 2012 [16], and confocal phase maps of biological cells were reported in a follow-up paper in 2013 by the same group [17] In these original proposals, a point-scanning system was adopted.
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