ZFIN ID: ZDB-PUB-191102-37
Dual-beam confocal light-sheet microscopy via flexible acousto-optic deflector
Gavryusev, V., Sancataldo, G., Ricci, P., Montalbano, A., Fornetto, C., Turrini, L., Laurino, A., Pesce, L., de Vito, G., Tiso, N., Vanzi, F., Silvestri, L., Pavone, F.S.
Date: 2019
Source: Journal of Biomedical Optics   24: 1-6 (Journal)
Registered Authors: Fornetto, Chiara, Tiso, Natascia, Turrini, Lapo, Vanzi, Francesco
Keywords: acousto-optic deflector, confocal detection, digital scanned laser light-sheet fluorescence microscopy, high contrast, high-throughput microscopy, light-sheet microscopy, mouse brain, zebrafish brain
MeSH Terms:
  • Animals
  • Brain/diagnostic imaging
  • Equipment Design
  • High-Throughput Screening Assays/methods
  • Image Processing, Computer-Assisted/methods*
  • Larva/cytology
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Confocal/instrumentation
  • Microscopy, Confocal/methods*
  • Microscopy, Fluorescence/instrumentation
  • Microscopy, Fluorescence/methods*
  • Zebrafish
PubMed: 31674164 Full text @ J. Biomed. Opt.
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 acousto-optic 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.