Active Remote Focus Stabilization in Oblique Plane Microscopy

Biomedical Optics Express, Journal Year: 2025, Volume and Issue: 16(3), P. 1216 - 1216

Published: Feb. 17, 2025

Light-sheet fluorescence microscopy (LSFM) has demonstrated great potential in the life sciences owing to its efficient volumetric imaging capabilities. For long-term imaging, light-sheet typically needs be stabilized detection focal plane for best results. Current stabilization methods rely on emission from sample, which may interrupt scientific and add sample photobleaching. Here, we show that oblique microscopes (OPM), a subset of LSFM where single primary objective is used illumination detection, can achieved without expending fluorescence. Our method achieves ∼21 nm axial precision maintains well within depth focus system hour-long acquisition runs lab environment would otherwise detune system. We demonstrate subcellular actin skeleton melanoma cancer cells with OPM.

Language: Английский

Single Objective Light Sheet Microscopy allows high-resolutioninvivo brain imaging of Drosophila.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 7, 2024

Abstract In vivo imaging of dynamic sub-cellular brain structures in Drosophila melanogaster is key to understanding several phenomena neuroscience. However, a trade-off between spatial resolution, speed, photodamage, and setup complexity limits its implementation. Here, we designed built single objective light sheet microscope, customized for adult flies optimized maximum resolution. Unlike multi-objective setups, the microscope uses at fly head interface, facilitating sample mounting inspection, reducing invasiveness. contrast two-photon microscopies, light-sheet configuration with visible excitation offers reduced phototoxicity. We demonstrate membrane, mitochondria, dense-core vesicles axonal projections small lateral ventral neurons. The achieved resolution was 380 500 nm within field view 70×50×12 μm 3 . This unique combination easy mounting, high low-phototoxicity illumination paves way new studies living flies.

Language: Английский

Active Remote Focus Stabilization in Oblique Plane Microscopy

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 2, 2024

Light-sheet fluorescence microscopy (LSFM) has demonstrated great potential in the life sciences owing to its efficient volumetric imaging capabilities. For long term imaging, light-sheet typically needs be stabilized detection focal plane for best results. Current stabilization methods rely on emission from sample, which may interrupt scientific and add sample photobleaching. Here we show that oblique microscopes (OPM), a subset of LSFM where single primary objective is used illumination detection, can achieved without expending fluorescence. Our method achieves ~43nm axial precision maintains well within depth focus system hour

Language: Английский