Journal of Biomedical Optics, Journal Year: 2024, Volume and Issue: 29(03)
Published: March 21, 2024
The reprojection setup typical of oblique plane microscopy (OPM) limits the effective aperture imaging system, and therefore its efficiency resolution. Large system is only possible through use custom specialized optics. A full-aperture OPM made with off shelf components would both improve performance method encourage widespread adoption.
Language: Английский
Nature Methods, Journal Year: 2025, Volume and Issue: unknown
Published: Feb. 12, 2025
Language: Английский
Citations
1
Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)
Published: March 29, 2024
Abstract Projection imaging accelerates volumetric interrogation in fluorescence microscopy, but for multi-cellular samples, the resulting images may lack contrast, as many structures and haze are summed up. Here, we demonstrate rapid pro jective light-sheet with p arameter s election (props) of depth, position viewing angle. This allows us to selectively image different sub-volumes a sample, rapidly switch between them exclude background fluorescence. Here power props by functional within distinct regions zebrafish brain, monitoring calcium firing inside muscle cells moving Drosophila larvae, super-resolution selected cell layers, optically unwrapping curved surface embryo. We anticipate that will accelerate interrogation, ranging from subcellular mesoscopic scales.
Language: Английский
Citations
6
Optica, Journal Year: 2023, Volume and Issue: 10(11), P. 1571 - 1571
Published: Oct. 18, 2023
Understanding the intricate interplay and inter-connectivity of biological processes across an entire organism is important in various fields biology, including cardiovascular research, neuroscience, developmental biology. Here, we present a mesoscopic oblique plane microscope (OPM) that enables whole imaging with high speed while revealing fine details such as endothelial nuclei. A microprism underneath sample enhances axial resolution optical sectioning through total internal reflection light sheet. Through rapid refocusing sheet, depth extended up to threefold keeping constant. Using low magnification objectives large field view, realize over volume 3.7mm×1.5mm×1mm ∼2.3µm lateral ∼9.2µm resolution. Applying OPM, demonstrate vivo toto zebrafish vasculature its nuclei, blood flow dynamics at 12 Hz acquisition rate, resulting quantitative map organism.
Language: Английский
Citations
12
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
Citations
0
Biomedical Optics Express, Journal Year: 2024, Volume and Issue: 15(8), P. 4498 - 4498
Published: June 28, 2024
Adaptive optics (AO) can restore diffraction-limited performance when imaging beyond superficial cell layers in vivo and vitro , as such, is of interest for advanced 3D microscopy methods such light-sheet fluorescence (LSFM). In a typical LSFM system, the illumination detection paths are separate subject to different optical aberrations. To achieve optimal microscope performance, it necessary sense correct these aberrations both light paths, resulting complex system. Here, we show that an oblique plane (OPM), type with single primary objective lens, same deformable mirror detection. Besides reducing complexity, AO OPM also restores relative alignment focal plane, projection mode stabilize improve wavefront correction sensorless format. We demonstrate on fluorescent nanospheres by vasculature cancer cells zebrafish embryos embedded glass capillary, restoring diffraction limited resolution improving signal strength twofold.
Language: Английский
Citations
3
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown
Published: March 2, 2024
Abstract Many biological processes span multiple time and length scales, including developmental cancer metastasis. While light-sheet fluorescence microscopy (LSFM) has become a fast efficient method for imaging of organisms, cells sub-cellular dynamics, simultaneous observations across these scales have remained challenging. Moreover, continuous high-resolution inside living organisms mostly been limited to few hours as regions interest quickly move out view due sample movement growth. Here, we present self-driving, multi-resolution microscope platform controlled by custom Python-based software, observe quantify dynamics entire in vitro vivo over imaging. We apply the study processes, invasion metastasis, provide quantitative multi-scale analysis immune-cancer cell interactions zebrafish xenografts.
Language: Английский
Citations
1
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown
Published: March 22, 2024
Abstract Adaptive optics (AO) can restore diffraction limited performance when imaging beyond superficial cell layers in vivo and vitro , as such is of interest for advanced 3D microscopy methods light-sheet fluorescence (LSFM). In a typical LSFM system, the illumination detection paths are separate subject to different optical aberrations. To achieve optimal microscope performance, it necessary sense correct these aberrations both light paths, resulting complex system. Here, we show that an oblique plane (OPM), type with single primary objective lens, same deformable mirror detection. Besides reducing complexity, AO OPM also restores relative alignment focal plane, projection mode stabilize improve wavefront correction sensorless format. We demonstrate on fluorescent nanospheres by vasculature cancer cells zebrafish embryos embedded glass capillary, restoring resolution improving signal strength twofold.
Language: Английский
Citations
0
Journal of Biomedical Optics, Journal Year: 2024, Volume and Issue: 29(08)
Published: Aug. 8, 2024
The technique of remote focusing (RF) has attracted considerable attention among microscopists due to its ability quickly adjust focus across different planes, thus facilitating quicker volumetric imaging. However, the difficulty in changing objectives align with a matching objective setting while upholding key requirements remains challenge.
Language: Английский
Citations
0
Biomedical Optics Express, Journal Year: 2024, Volume and Issue: 15(12), P. 6715 - 6715
Published: Sept. 12, 2024
Rapid three-dimensional imaging over extended fields of view (FOVs) is crucial to the study organism-wide systems and biological processes
Language: Английский
Citations
0
npj Imaging, Journal Year: 2024, Volume and Issue: 2(1)
Published: Nov. 21, 2024
Single-molecule localization microscopy has revealed cellular architectures and molecular dynamics beyond the diffraction limit of light. However, imaging thick samples presents challenges from increased fluorescence background. Light sheet illumination, which utilizes a plane light for optical sectioning, is effective in reducing background, photobleaching, photodamage. Here, we present principles single-molecule followed by an introduction to geometries their applications. Finally, discuss illumination approaches high- super-resolution biological structures dynamics.
Language: Английский
Citations
0