Elimination of imaging artifacts in second harmonic generation microscopy using interferometry

Pinsard, Maxime; Schmeltz, Margaux; Kolk, Jarno van der; Patten, Shunmoogum A.; Ibrahim, Heide; Ramunno, Lora; Schanne-Klein, Marie-Claire; Légaré, François

doi:10.1364/boe.10.003938

Cited by 7 publications

(5 citation statements)

References 51 publications

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“…In the 4-view MVDA, a striping effect is visible. This striping has the direction of the z-axis of each individual view and is due to the incomplete removal of the PSF elongation after deconvolution caused by strong local constructive interference effects observed in such tissues 28 , 29 . The addition of more views minimizes this effect, and individual fibrils become better visible (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Multiview deconvolution approximation multiphoton microscopy of tissues and zebrafish larvae

Kapsokalyvas

Rosas

Janssen

et al. 2021

Sci Rep

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Imaging in three dimensions is necessary for thick tissues and small organisms. This is possible with tomographic optical microscopy techniques such as confocal, multiphoton and light sheet microscopy. All these techniques suffer from anisotropic resolution and limited penetration depth. In the past, Multiview microscopy—imaging the sample from different angles followed by 3D image reconstruction—was developed to address this issue for light sheet microscopy based on fluorescence signal. In this study we applied this methodology to accomplish Multiview imaging with multiphoton microscopy based on fluorescence and additionally second harmonic signal from myosin and collagen. It was shown that isotropic resolution was achieved, the entirety of the sample was visualized, and interference artifacts were suppressed allowing clear visualization of collagen fibrils and myofibrils. This method can be applied to any scanning microscopy technique without microscope modifications. It can be used for imaging tissue and whole mount small organisms such as heart tissue, and zebrafish larva in 3D, label-free or stained, with at least threefold axial resolution improvement which can be significant for the accurate quantification of small 3D structures.

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Section: Resultsmentioning

confidence: 99%

“…5 ). Interferometric second harmonic generation microscopy 28 , 29 has been proposed to eliminate interference artifacts with very promising results. Indeed, such artifacts are completely suppressed offering a clear view of the inherent SHG signal, although this technique is very sensitive, it is limited to thin tissue sections.…”

Section: Discussionmentioning

confidence: 99%

Multiview deconvolution approximation multiphoton microscopy of tissues and zebrafish larvae

Kapsokalyvas

Rosas

Janssen

et al. 2021

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Interferometric second harmonic generation microscopy 25,26 has been proposed to eliminate interference artifacts with very promising results. Indeed, such artifacts are completely suppressed offering a clear view of the inherent SHG signal, although since this technique is very sensitive it is limited to thin tissue sections.…”

Section: Discussionmentioning

confidence: 99%

“…In the 4-view MVD, a striping effect is visible. This striping has the direction of the z-axis of each individual view and is due to the incomplete removal of the PSF elongation after deconvolution caused by strong local constructive interference effects observed in such tissues 25,26 . The addition of more views minimizes this effect, and individual brils become better visible (Fig.…”

Section: Multiview Deconvolution Of Collagen Shg With 4 Views and 8 Vmentioning

confidence: 99%

Multiview Deconvolution Two-photon Laser Scanning Microscopy

Kapsokalyvas

Rosas

Janssen

et al. 2020

Preprint

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Imaging in three dimensions is necessary for thick tissues and small organisms. This is possible with tomographic optical microscopy techniques such as confocal, two-photon and light sheet microscopy. All these techniques suffer from anisotropic resolution and limited penetration depth. In the past, Multiview microscopy - imaging the sample from different angles followed by 3D image reconstruction - was developed to address this issue for light sheet microscopy based on fluorescence signal. In this study we applied this methodology to accomplish Multiview imaging with two-photon microscopy based on fluorescence and additionally second harmonic signal from myosin and collagen. It was shown that isotropic resolution was achieved, the entirety of the sample was visualized, and interference artifacts were suppressed allowing clear visualization of collagen fibrils and myofibrils. This method can be applied to any scanning microscopy technique without microscope modifications. It can be used for imaging tissue and whole mount small organisms such as heart tissue, and zebrafish larva in 3D, label-free or stained, with at least 3-fold axial resolution improvement which can be significant for the accurate quantification of small 3D structures.

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“…With the effect of the background noise of the detection elements or pixelation of the sampling signal, the experimental lateral resolution may be larger than the theoretical value in the actual system. [ 43 ] In our system, the lateral SHG resolution is approximately 400 nm.…”

Section: Instrumentationmentioning

confidence: 99%

High‐Throughput Scanning Second‐Harmonic‐Generation Microscopy for Polar Materials

et al. 2023

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The Materials Genome Initiative aims to discover, develop, manufacture, and deploy advanced materials at twice the speed of conventional approaches. To achieve this, high‐throughput characterization is essential for the rapid screening of candidate materials. In this study, a high‐throughput scanning second‐harmonic‐generation microscope with automatic partitioning, accurate positioning, and fast scanning is developed that can rapidly probe and screen polar materials. Using this technique, typical ferroelectrics, including periodically poled lithium niobate crystals and PbZr0.2Ti0.8O3 (PZT) thin films are first investigated, whereby the microscopic domain structures are clearly revealed. This technique is then applied to a compositional‐gradient (100−x)%BaTiO3−x%SrTiO3 film and a thickness‐gradient PZT film to demonstrate its high‐throughput capabilities. Since the second‐harmonic‐generation signal is correlated with the macroscopic remnant polarization over the probed region determined by the laser spot, it is free of artifacts arising from leakage current and electrostatic interference, while materials’ symmetries and domain structures must be carefully considered in the data analysis. It is believed that this work can help promote the high‐throughput development of polar materials and contribute to the Materials Genome Initiative.

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Elimination of imaging artifacts in second harmonic generation microscopy using interferometry

Cited by 7 publications

References 51 publications

Multiview deconvolution approximation multiphoton microscopy of tissues and zebrafish larvae

Multiview deconvolution approximation multiphoton microscopy of tissues and zebrafish larvae

Multiview Deconvolution Two-photon Laser Scanning Microscopy

High‐Throughput Scanning Second‐Harmonic‐Generation Microscopy for Polar Materials

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