Abstract
Light sheet fluorescence microscopy (LSFM) is a powerful tool for investigating model organisms including zebrafish. However, due to scattering and refractive index variations within the sample, the resulting image often suffers from low contrast. Structured illumination (SI) has been combined with scanned LSFM to remove out-of-focus and scattered light using square-law detection. Here, we demonstrate that the combination of LSFM with linear reconstruction SI can further increase resolution and contrast in the vertical and axial directions compared to the widely adopted root-mean square reconstruction method while using the same input images. We apply this approach to imaging neural activity in 7-day postfertilization zebrafish larvae. We imaged two-dimensional sections of the zebrafish central nervous system in two colors at an effective frame rate of 7 frames per second.
Highlights
A fundamental problem in biomedical science is understanding the three-dimensional (3-D) structure of the brain and other organs at the cellular level
We evaluate the performance of our digitally scanned light sheet microscopy (DSLM) system comparing root mean square (RMS)-Structured illumination (SI) and linear reconstruction structured illumination (LR-SI) with different parameters, imaging fluorescent bead phantoms and thick biological samples and compare the contrast enhancement
3 Results and Discussion In Light sheet fluorescence microscopy (LSFM), the thickness of the sheet is directly related to the optical sectioning performance of the system
Summary
A fundamental problem in biomedical science is understanding the three-dimensional (3-D) structure of the brain and other organs at the cellular level. Distinct from the widely used epi-illumination widefield (WF) microscope or laser scanning confocal microscope, the planar illumination scheme of the LSFM allows researchers to study biological samples in three dimensions with significantly less light exposure and drastically lower illumination intensity,[4] while still allowing for improved optical sectioning. What makes this possible is the sheet-like illumination of the sample at the focal plane of the detection objective lens, which greatly suppresses out-of-focus light. Because of its WF detection scheme, LSFM has a much higher temporal resolution over a large field-of-view (FoV) compared with laser point scanning techniques, such as confocal microscopy[5] and two-photon microscopy.[6]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
