Optical Sectioning Microscopy Through Single-Shot Lightfield Protocol

Sánchez-Ortiga, Emilio; Scrofani, Gabriele; Saavedra, Genaro; Martı́nez-Corral, Manuel

doi:10.1109/access.2020.2966323

Cited by 19 publications

(10 citation statements)

References 36 publications

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“…To reconstruct the sample volume, we used the shift and multiply (S&M) method [ 22 ], which is capable of providing optical sectioning in real time. Furthermore, this algorithm avoids the background noise typically provided by deconvolution methods.…”

Section: Resultsmentioning

confidence: 99%

The Lightfield Microscope Eyepiece

Incardona

Tolosa

Scrofani

et al. 2021

Sensors

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Lightfield microscopy has raised growing interest in the last few years. Its ability to get three-dimensional information about the sample in a single shot makes it suitable for many applications in which time resolution is fundamental. In this paper we present a novel device, which is capable of converting any conventional microscope into a lightfield microscope. Based on the Fourier integral microscope concept, we designed the lightfield microscope eyepiece. This is coupled to the eyepiece port, to let the user exploit all the host microscope’s components (objective turret, illumination systems, translation stage, etc.) and get a 3D reconstruction of the sample. After the optical design, a proof-of-concept device was built with off-the-shelf optomechanical components. Here, its optical performances are demonstrated, which show good matching with the theoretical ones. Then, the pictures of different samples taken with the lightfield eyepiece are shown, along with the corresponding reconstructions. We demonstrated the functioning of the lightfield eyepiece and lay the foundation for the development of a commercial device that works with any microscope.

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Section: Resultsmentioning

confidence: 99%

The Lightfield Microscope Eyepiece

Incardona

Tolosa

Scrofani

et al. 2021

Sensors

Self Cite

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“…In Fig. 2 an example of the results that can be achieved with the application of the new optical-sectioning algorithm above mentioned 7 is shown. Following the scheme shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The integral image of a 3D sample consisting of a number of fluorescent beads of sizes ranging for 10 to 100 µm in water was obtained. A set of 25 refocused depth planes was obtained after the application of the conventional refocusing (CR) algorithm 3 , the deconvolution-based refocusing 6 (DBR) scheme, and the optical-sectioning refocusing 7 (OSR) technique. After the application of a maximum intensity projection algorithm, three 3D renders were obtained.…”

Section: Resultsmentioning

confidence: 99%

“…This algorithm, however, had the drawback of requiring long calculation time, and of providing important background noise typically inherent to deconvolution procedures. Recently, a new protocol for obtaining optically-sectioned 3D images in real time has been reported 7 . This procedure overcomes the drawbacks described above, and it is well suited for the case of fluorescent, sparse, 3D microscopic samples.…”

Section: Optical-sectioning Imaging By Fimicmentioning

confidence: 99%

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Fourier-domain lightfield microscopy: a new paradigm in 3D microscopy

Sánchez-Ortiga

Scrofani

Martı́nez-Corral

et al. 2020

Biomedical Imaging and Sensing Conference 2020

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Recently, integral (also known as lightfield or plenoptic) imaging concept has been applied successfully to microscopy. The main advantage of lightfield microscopy when compared with conventional 3D imaging techniques is that it offers the possibility of capturing the 3D information of the sample after a single shot. However, integral microscopy is now facing many challenges, like improving the resolution and depth of field of the reconstructed specimens or the development and optimization of specially-adapted reconstruction algorithms. This contribution is devoted to review a new paradigm in lightfield microscopy, namely, the Fourier-domain integral microscope (FiMic), that improves the capabilities of the technique, and to present recent advances and applications of this new architecture.

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“…Existing, optimized, algorithms [23,24] can be utilized to estimate the location of the centre of each foci with a precision much finer than its diffraction-limited width. It is well established that information captured within different microlenses can be combined to localise objects in 3D [25,26]. We demonstrate that the temporal sparsity of SMLM, which limits the probability of overlap between diffraction-limited images of distinct emitters, makes it an extremely attractive technique to combine with light field microscopy.…”

Section: Introductionmentioning

confidence: 94%

Single molecule light field microscopy

Sims¹,

Rehman²,

Lenz³

et al. 2020

Biophotonics and Biomedical Microscopy

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We introduce single molecule light field microscopy (SMLFM), a novel 3D single molecule localization technique that is capable of up to 20 nm lateral and axial precision across a 6 µm depth of field. SMLFM can be readily implemented by installing a refractive microlens array into the conjugate back focal plane of any widefield single molecule localization system. We demonstrate that 3D localization can be performed by post-processing 2D localization data generated by common, widely-used, algorithms. In this work we benchmark the performance of SMLFM and finally showcase its capabilities by imaging fluorescently labeled membranes of fixed eukaryotic cells below the diffraction limit.

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Optical Sectioning Microscopy Through Single-Shot Lightfield Protocol

Cited by 19 publications

References 36 publications

The Lightfield Microscope Eyepiece

The Lightfield Microscope Eyepiece

Fourier-domain lightfield microscopy: a new paradigm in 3D microscopy

Single molecule light field microscopy

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