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RESEARCH PRODUCT
Dual-beam confocal light-sheet microscopy via flexible acousto-optic deflector
Luca PesceChiara FornettoLapo TurriniGiuseppe VitoFrancesco S. PavoneLudovico SilvestriPietro RicciNatascia TisoGiuseppe SancataldoAnnunziatina LaurinoFrancesco VanziAlberto MontalbanoVladislav Gavryusevsubject
Image formationPaperMaterials scienceImage qualityConfocalBiomedical Engineeringacousto-optic deflector; confocal detection; digital scanned laser light-sheet fluorescence microscopy; high contrast; high-throughput microscopy; light-sheet microscopy; mouse brain; zebrafish brainconfocal detection01 natural scienceslaw.invention010309 opticsBiomaterialsMiceacousto-optic deflectorOpticslaw0103 physical sciencesMicroscopyImage Processing Computer-AssistedAnimalsZebrafishhigh-throughput microscopyconfocal light-sheet microscopyMicroscopyMicroscopy Confocalbusiness.industryhigh contrastRolling shutterBrainEquipment DesignLaserFrame ratezebrafish brainAtomic and Molecular Physics and OpticsElectronic Optical and Magnetic MaterialsHigh-Throughput Screening AssaysMice Inbred C57BLdigital scanned laser light-sheet fluorescence microscopyMicroscopy FluorescenceLight sheet fluorescence microscopyLarvamouse brainbusinesslight-sheet microscopydescription
Confocal detection in digital scanned laser light-sheet fluorescence microscopy (DSLM) has been established as a gold standard method to improve image quality. The selective line detection of a complementary metal-oxide-semiconductor camera (CMOS) working in rolling shutter mode allows the rejection of out-of-focus and scattered light, thus reducing background signal during image formation. Most modern CMOS have two rolling shutters, but usually only a single illuminating beam is used, halving the maximum obtainable frame rate. We report on the capability to recover the full image acquisition rate via dual confocal DSLM by using an acoustooptic deflector. Such a simple solution enables us to independently generate, control and synchronize two beams with the two rolling slits on the camera. We show that the doubling of the imaging speed does not affect the confocal detection high contrast. (C) The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2019-05-13 | Journal of Biomedical Optics |