Antoine Hubert, Mattéo Dommanget-Kott, Sébastien Wolf, Thomas Panier, G. Debrégeas, Volker Bormuth
We describe a 3D holographic two-photon optogenetics setup based on acousto-optic deflectors combined with a custom-made compact light-sheet microscope setup. The system allows performing 3D-targeted perturbations of neuronal activity while simultaneously imaging the whole brain neuronal response over large field-of-view. This random-access holographic light patterning method offers high-speed sequential activation of dense neuronal assemblies spread over large areas while maintaining constant lateral and axial resolution. Here, we report on a systematic characterization of the stimulated neurons response and perform functional imaging on zebrafish (Danio Rerio) larvae while stimulating single neurons.
{"title":"Random-access two-photon holographic optogenetic stimulation combined with brain-wide functional light-sheet imaging in larval zebrafish","authors":"Antoine Hubert, Mattéo Dommanget-Kott, Sébastien Wolf, Thomas Panier, G. Debrégeas, Volker Bormuth","doi":"10.1117/12.2671030","DOIUrl":"https://doi.org/10.1117/12.2671030","url":null,"abstract":"We describe a 3D holographic two-photon optogenetics setup based on acousto-optic deflectors combined with a custom-made compact light-sheet microscope setup. The system allows performing 3D-targeted perturbations of neuronal activity while simultaneously imaging the whole brain neuronal response over large field-of-view. This random-access holographic light patterning method offers high-speed sequential activation of dense neuronal assemblies spread over large areas while maintaining constant lateral and axial resolution. Here, we report on a systematic characterization of the stimulated neurons response and perform functional imaging on zebrafish (Danio Rerio) larvae while stimulating single neurons.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"70 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130606899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
J. Ferrer Ortas, P. Mahou, S. Escot, C. Stringari, Nicolas B. David, Laure Bally-Cuif, Nicolas Dray, Michel Négrerie, Willy Supatto, E. Beaurepaire
We present color third-order sum-frequency generation (color TSFG) microscopy, a multiphoton imaging strategy based on the simultaneous detection of several third-order coherent signals produced by two synchronized femtosecond pulse trains. We demonstrate that it can be used to obtain red blood cell (RBC)-specific label-free contrast in live zebrafish and is a promising tool for probing RBC oxygenation.
{"title":"Color TSFG microscopy of red blood cells and oxygenation","authors":"J. Ferrer Ortas, P. Mahou, S. Escot, C. Stringari, Nicolas B. David, Laure Bally-Cuif, Nicolas Dray, Michel Négrerie, Willy Supatto, E. Beaurepaire","doi":"10.1117/12.2670229","DOIUrl":"https://doi.org/10.1117/12.2670229","url":null,"abstract":"We present color third-order sum-frequency generation (color TSFG) microscopy, a multiphoton imaging strategy based on the simultaneous detection of several third-order coherent signals produced by two synchronized femtosecond pulse trains. We demonstrate that it can be used to obtain red blood cell (RBC)-specific label-free contrast in live zebrafish and is a promising tool for probing RBC oxygenation.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"815 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131551183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Optical imaging is a non-invasive technique that is able to monitor hemodynamic and metabolic responses during neurosurgery. However, a robust quantification is complicated to perform. To overcome this issue, phantoms that mimic biological tissues are required for the development of imaging systems in order to reach a true standardization. In this work, we explore the possibility to use a combined liquid blood phantom with cytochrome contained yeast to evaluate the reliability of hyperspectral imaging to measure oxygenation and metabolic changes. This phantom can be used to verify the reliability of intraoperative optical setups before moving on to clinical application.
{"title":"Evaluation of hyperspectral imaging measurements of changes in hemoglobin oxygenation and oxidation of cytochrome-c-oxidase using a liquid blood phantom","authors":"C. Caredda, F. Lange, I. Tachtsidis, B. Montcel","doi":"10.1117/12.2670746","DOIUrl":"https://doi.org/10.1117/12.2670746","url":null,"abstract":"Optical imaging is a non-invasive technique that is able to monitor hemodynamic and metabolic responses during neurosurgery. However, a robust quantification is complicated to perform. To overcome this issue, phantoms that mimic biological tissues are required for the development of imaging systems in order to reach a true standardization. In this work, we explore the possibility to use a combined liquid blood phantom with cytochrome contained yeast to evaluate the reliability of hyperspectral imaging to measure oxygenation and metabolic changes. This phantom can be used to verify the reliability of intraoperative optical setups before moving on to clinical application.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"18 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124278686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Berfin Arli, O. Dinc, Merve Türker-Burhan, S. Tozburun
The potential of laser-induced thermal therapy can be reassessed in treating abnormal mucosal tissues with advances in fiber optics, diode laser technology, and optical imaging modalities. In this context, studies optimizing a large parameter matrix (e.g., laser power, surface scanning speed, beam diameter, and irradiation duration) may be of interest. This study presents an artificial intelligence algorithm utilizing a generative adversarial network that predicts dark-field microscopy images from bright-field images of H&E-stained esophageal specimens. The calculated structural similarity index measurement between ground truth and the predicted dark-field image reaches an average of 74%. Also, the mean squared error is 0.7%.
{"title":"Predicting dark-field images of H and E-stained esophageal specimens","authors":"Berfin Arli, O. Dinc, Merve Türker-Burhan, S. Tozburun","doi":"10.1117/12.2672202","DOIUrl":"https://doi.org/10.1117/12.2672202","url":null,"abstract":"The potential of laser-induced thermal therapy can be reassessed in treating abnormal mucosal tissues with advances in fiber optics, diode laser technology, and optical imaging modalities. In this context, studies optimizing a large parameter matrix (e.g., laser power, surface scanning speed, beam diameter, and irradiation duration) may be of interest. This study presents an artificial intelligence algorithm utilizing a generative adversarial network that predicts dark-field microscopy images from bright-field images of H&E-stained esophageal specimens. The calculated structural similarity index measurement between ground truth and the predicted dark-field image reaches an average of 74%. Also, the mean squared error is 0.7%.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"5 2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130684347","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Demián Augusto Vera, H. García, G. Baez, M. Waks-Serra, N. Carbone, D. Iriarte, J. Pomarico
In this work we introduce an analytical way of computing the photon measurement density functions in multilayered flat and spherical media. Comparisons with Monte Carlo simulations in the particular case of two-layered media show very good agreement (differences below 10%), with the additional advantage that the time taken by the theoretical calculations is several orders of magnitude (more than 6) lower than the corresponding Monte Carlo calculations.
{"title":"Analytical photon measurement density functions in flat and spherical layered media","authors":"Demián Augusto Vera, H. García, G. Baez, M. Waks-Serra, N. Carbone, D. Iriarte, J. Pomarico","doi":"10.1117/12.2667099","DOIUrl":"https://doi.org/10.1117/12.2667099","url":null,"abstract":"In this work we introduce an analytical way of computing the photon measurement density functions in multilayered flat and spherical media. Comparisons with Monte Carlo simulations in the particular case of two-layered media show very good agreement (differences below 10%), with the additional advantage that the time taken by the theoretical calculations is several orders of magnitude (more than 6) lower than the corresponding Monte Carlo calculations.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128141394","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ankit Butola, Sebastián Acuña, D. H. Hansen, Krishna Agarwal
Structured illumination microscopy (SIM) is a most popular super-resolution technique used in cell biology and bio-imaging. Here, we present a novel approach to realize multiscale super-resolution SIM by swapping the non-linearity between instrumentation and reconstruction algorithm to achieve super-resolution. Our goal is to overcome two conventional limitations of SIM i.e., fixed resolution and the need of precise knowledge of illumination pattern. The optical system encodes higher order frequencies of the sample by projecting PSF-modulated binary patterns for illuminating the sample plane, which do not have clean Fourier peaks conventionally used in SIM. These patterns fold high frequency content of sample into the measurements in an obfuscated manner, which are de-obfuscated using multiple signal classification algorithm. Our approach eliminates the need of clean peaks in the illumination pattern, which have multiple advantages i.e., simple instrumentation and the flexibility of using different collection lenses. The reconstruction algorithm used in the proposed work does not require known illumination. Finally, we reduce the sensitivity of reconstruction algorithm to the signal to background ratio. Here, we acquired patterned illumination images of the same sample using different collection objective lenses, and obtained diffraction limited as well as super-resolved images, supporting 4 different resolution in the same system through SIM. Our experimental results with multiple collection objective lens show wider applicability of the proposed system at signal to background ration as small as <3.
{"title":"Multiscale resolution in structured illumination microscopy","authors":"Ankit Butola, Sebastián Acuña, D. H. Hansen, Krishna Agarwal","doi":"10.1117/12.2671708","DOIUrl":"https://doi.org/10.1117/12.2671708","url":null,"abstract":"Structured illumination microscopy (SIM) is a most popular super-resolution technique used in cell biology and bio-imaging. Here, we present a novel approach to realize multiscale super-resolution SIM by swapping the non-linearity between instrumentation and reconstruction algorithm to achieve super-resolution. Our goal is to overcome two conventional limitations of SIM i.e., fixed resolution and the need of precise knowledge of illumination pattern. The optical system encodes higher order frequencies of the sample by projecting PSF-modulated binary patterns for illuminating the sample plane, which do not have clean Fourier peaks conventionally used in SIM. These patterns fold high frequency content of sample into the measurements in an obfuscated manner, which are de-obfuscated using multiple signal classification algorithm. Our approach eliminates the need of clean peaks in the illumination pattern, which have multiple advantages i.e., simple instrumentation and the flexibility of using different collection lenses. The reconstruction algorithm used in the proposed work does not require known illumination. Finally, we reduce the sensitivity of reconstruction algorithm to the signal to background ratio. Here, we acquired patterned illumination images of the same sample using different collection objective lenses, and obtained diffraction limited as well as super-resolved images, supporting 4 different resolution in the same system through SIM. Our experimental results with multiple collection objective lens show wider applicability of the proposed system at signal to background ration as small as <3.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"59 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130473287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Demián Augusto Vera, H. García, N. Carbone, M. Waks-Serra, D. Iriarte, J. Pomarico
In this work we derive general equations for the analytical calculation of photon mean partial pathlengths (MPPLs) in turbid media with an arbitrary number of layers. Comparisons with their Monte Carlo (MC) counterpart show excellent agreement. These quantities can now be used to retrieve haemoglobin concentrations changes in cerebral blood in real-time and with minimal computing requirements.
{"title":"Theoretical computation of photon mean partial pathlengths in multilayered turbid media","authors":"Demián Augusto Vera, H. García, N. Carbone, M. Waks-Serra, D. Iriarte, J. Pomarico","doi":"10.1117/12.2670453","DOIUrl":"https://doi.org/10.1117/12.2670453","url":null,"abstract":"In this work we derive general equations for the analytical calculation of photon mean partial pathlengths (MPPLs) in turbid media with an arbitrary number of layers. Comparisons with their Monte Carlo (MC) counterpart show excellent agreement. These quantities can now be used to retrieve haemoglobin concentrations changes in cerebral blood in real-time and with minimal computing requirements.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"13 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127557158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K. Tomita, S. Makita, N. Fukutake, R. Morishita, I. Abd El-Sadek, P. Mukherjee, Antonia Lichtenegger, Y. Yasuno
A new formulation of Optical Coherence Tomography (OCT) is presented. This formulation can represent a meaningful OCT image and speckles as two independent mathematical entities. In addition, by using the same formulation strategy, a new imaging modality to generate a spatially differential image, similar to differential interferometric contrast microscopy is demonstrated.
{"title":"New formulation of OCT for analytical signal-to-speckle separation and volumetric differential contrast imaging","authors":"K. Tomita, S. Makita, N. Fukutake, R. Morishita, I. Abd El-Sadek, P. Mukherjee, Antonia Lichtenegger, Y. Yasuno","doi":"10.1117/12.2670518","DOIUrl":"https://doi.org/10.1117/12.2670518","url":null,"abstract":"A new formulation of Optical Coherence Tomography (OCT) is presented. This formulation can represent a meaningful OCT image and speckles as two independent mathematical entities. In addition, by using the same formulation strategy, a new imaging modality to generate a spatially differential image, similar to differential interferometric contrast microscopy is demonstrated.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"12632 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130074078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In this paper, we present a Wavefront Division (WD) Fourier domain Swept Source Optical Coherence Tomography (SS-OCT) configuration. The presented WD-SS-OCT configuration has two key features that are discussed in this contribution. First, there is the possibility of variable splitting and flexible adjustment of the splitting ratio between the sample and the reference. This enables an adaptation to differing reflective and scattering features of various samples. The second feature is related to the increased throughput of the interferometer design. The use of polarization optics makes it possible to almost eliminate back-reflection losses completely to the source (up to 50% of the intensity in standard amplitude division-based OCT). WD-OCT configuration has its challenges compared to conventional OCT systems based on amplitude splitting. Nevertheless, the proposed design has also specific options and flexibilities that are worth to be shared with the OCT community.
{"title":"Wavefront division swept source OCT","authors":"B. Heise, Gopika Venugopal, I. Zorin, A. Prylepa","doi":"10.1117/12.2670763","DOIUrl":"https://doi.org/10.1117/12.2670763","url":null,"abstract":"In this paper, we present a Wavefront Division (WD) Fourier domain Swept Source Optical Coherence Tomography (SS-OCT) configuration. The presented WD-SS-OCT configuration has two key features that are discussed in this contribution. First, there is the possibility of variable splitting and flexible adjustment of the splitting ratio between the sample and the reference. This enables an adaptation to differing reflective and scattering features of various samples. The second feature is related to the increased throughput of the interferometer design. The use of polarization optics makes it possible to almost eliminate back-reflection losses completely to the source (up to 50% of the intensity in standard amplitude division-based OCT). WD-OCT configuration has its challenges compared to conventional OCT systems based on amplitude splitting. Nevertheless, the proposed design has also specific options and flexibilities that are worth to be shared with the OCT community.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"68 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127115943","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
S. Tomczewski, P. Węgrzyn, D. Borycki, K. Liżewski, M. Wielgo, A. Curatolo, M. Wojtkowski
We use Spatio-Temporal Optical Coherence Tomography (STOC-T) to record flicker ORG signals. The chirped frequency stimulus (5 Hz to 45 Hz) provides a more convenient way of measuring photoreceptors' frequency characteristics. Our method detected significant differences in response amplitudes in two measured subjects when comparing responses to different stimulus amplitudes as well as when comparing responses to rectangular and sinusoidal stimulation.
{"title":"Chirped frequency flicker optoretinography (fORG) with STOC-T","authors":"S. Tomczewski, P. Węgrzyn, D. Borycki, K. Liżewski, M. Wielgo, A. Curatolo, M. Wojtkowski","doi":"10.1117/12.2670862","DOIUrl":"https://doi.org/10.1117/12.2670862","url":null,"abstract":"We use Spatio-Temporal Optical Coherence Tomography (STOC-T) to record flicker ORG signals. The chirped frequency stimulus (5 Hz to 45 Hz) provides a more convenient way of measuring photoreceptors' frequency characteristics. Our method detected significant differences in response amplitudes in two measured subjects when comparing responses to different stimulus amplitudes as well as when comparing responses to rectangular and sinusoidal stimulation.","PeriodicalId":278089,"journal":{"name":"European Conference on Biomedical Optics","volume":"13 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127143081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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