In this work, a practical guide for the design of a Fourier lightfield microscope is reported. The fundamentals of the Fourier lightfield are presented and condensed on a set of contour plots from which the user can select the design values of the spatial resolution, the field of view, and the depth of field, as function of the specifications of the hardware of the host microscope. This work guides the reader to select the parameters of the infinity-corrected microscope objective, the optical relay lenses, the aperture stop, the microlens array, and the digital camera. A user-friendly graphic calculator is included to ease the design, even to those who are not familiar with the lightfield technology. The guide is aimed to simplify the design process of a Fourier lightfield microscope, which sometimes could be a daunting task, and in this way, to invite the widespread use of this technology. An example of a design and experimental results on imaging different types of samples is also presented.
In this work, the design, building, and testing of the most portable, easy-to-build, robust, handheld, and cost-effective Fourier Lightfield Microscope (FLMic) to date is reported. The FLMic is built by means of a surveillance camera lens and additional off-the-shelf optical elements, resulting in a cost-effective FLMic exhibiting all the regular sought features in lightfield microscopy, such as refocusing and gathering 3D information of samples by means of a single-shot approach. The proposed FLMic features reduced dimensions and light weight, which, combined with its low cost, turn the presented FLMic into a strong candidate for in-field application where 3D imaging capabilities are pursued. The use of cost-effective optical elements has a relatively low impact on the optical performance, regarding the figures dictated by the theory, while its price can be at least 100 times lower than that of a regular FLMic. The system operability is tested in both bright-field and fluorescent modes by imaging a resolution target, a honeybee wing, and a knot of dyed cotton fibers.
The resolution limit achievable with an optical system is a fundamental piece of information when characterizing its performance, mainly in case of microscopy imaging. Usually this information is given in the form of a distance, often expressed in microns, or in the form of a cutoff spatial frequency, often expressed in line pairs per mm. In modern imaging systems, where the final image is collected by pixelated digital cameras, the resolution limit is determined by the performance of both, the optical systems and the digital sensor. Usually, one of these factors is considered to be prevalent over the other for estimating the spatial resolution, leading to the global performance of the imaging system ruled by either the classical Abbe resolution limit, based on physical diffraction, or by the Nyquist resolution limit, based on the digital sensor features. This estimation fails significantly to predict the global performance of opto-digital imaging systems, like 3D microscopes, where none of the factors is negligible. In that case, which indeed is the most common, neither the Abbe formula nor the Nyquist formula provide by themselves a reliable prediction for the resolution limit. This is a serious drawback since systems designers often use those formulae as design input parameters. Aiming to overcome this lack, a simple mathematical expression obtained by finely articulating the Abbe and Nyquist formulas, to easily predict the spatial resolution limit of opto-digital imaging systems, is proposed here. The derived expression is tested experimentally, and shows to be valid in a broad range of opto-digital combinations.
We report a miniature Fourier Lightfield Microscope, based on a cost-effective reversed surveillance camera lens and a contact field stop. The miniscope captures efficiently 3D information, in fluorescence or brightfield mode, in one single shot.
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