Abstract
We demonstrate experimentally a scanning confocal microscopy technique based on digital holography. The method relies on digital holographic recording of the scanned spot. The data collected in this way contains all the necessary information to digitally produce three-dimensional images. Several methods to treat the data are presented. Examples of reflection and transmission images of epithelial cells and mouse brain tissue are shown.
Highlights
Confocal microscopy is a well established optical imaging method because of its threedimensional sectioning capability [1, 2]
The original proposal by Minsky relied on linear scattering and absorption of the sample as the contrast mechanism, confocal microscopy has found more widespread use in fluorescence imaging
Several techniques have been proposed to improve the image quality by reducing the scattering of the tissue by matching the refractive index with specific clearing agents [3, 4] or through adaptive optics, which can improve the quality of the probing beam and correct for sample-induced distortion [5, 6]
Summary
Confocal microscopy is a well established optical imaging method because of its threedimensional sectioning capability [1, 2]. The original proposal by Minsky relied on linear scattering and absorption of the sample as the contrast mechanism, confocal microscopy has found more widespread use in fluorescence imaging. The advantage of fluorescence confocal imaging is that fluorophores can label specific biological targets and that it provides a strong contrast mechanisms that does not rely on scattering. Several techniques have been proposed to improve the image quality by reducing the scattering of the tissue by matching the refractive index with specific clearing agents [3, 4] or through adaptive optics, which can improve the quality of the probing beam and correct for sample-induced distortion [5, 6].
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