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Abstract
Imaging specimens over large scales and with a sub-micron resolution is instrumental to biomedical research. Yet, the number of pixels to form such an image usually exceeds the number of pixels provided by conventional cameras. Although most microscopes are equipped with a motorized stage to displace the specimen and acquire the image tile-by-tile, we propose an alternative strategy that does not require to move any part in the sample plane. We propose to add a scanning mechanism in the detection unit of the microscope to collect sequentially different sub-areas of the field of view. Our approach, called remote scanning, is compatible with all camera-based microscopes. We evaluate the performances in both wide-field microscopy and full-field optical coherence tomography and we show that a field of view of 2.2 × 2.2 mm2 with a 1.1 μm resolution can be acquired. We finally demonstrate that the method is especially suited to image motion-sensitive samples and large biological samples such as millimetric engineered tissues.
Highlights
Deciphering biological processes often relies on the dual ability to perform in toto threedimensional imaging of the sample, which can be of macroscopic size, and to reach a sub-cellular (i.e. ∼ μm) resolution
We all experienced that a high magnification microscope objective (MO) provides a small field of view (FOV) at high lateral resolution, while a low magnification MO would expand the FOV but with a degraded resolution
We report on a simple method to significantly increase the FOV of a camera-based microscope without moving the sample in the focal plane of the MO
Summary
Deciphering biological processes often relies on the dual ability to perform in toto threedimensional imaging of the sample, which can be of macroscopic size, and to reach a sub-cellular (i.e. ∼ μm) resolution. The spatial bandwidth product (SBP) characterizes the throughput of an objective. It is defined as the number of pixels necessary to capture the full FOV at Nyquist sampling [1,2,3]. A standard X10 with a 1.1 μm resolution and a theoretical 2.65 × 2.65 mm FOV has an SBP of 21 megapixels. This number already exceeds the typical number of pixels on a camera, commonly between 1 and 4 megapixels, meaning that detectors are the limiting factor and that most of the information captured and transmitted by a MO is discarded
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