Multifocal structured illumination fluorescence microscopy with large field-of-view and high spatio-temporal resolution

Light-sheet microscopy has become the preferred method for long-term imaging of large living samples because of its low photo-invasiveness and good optical sectioning capabilities. Unfortunately, refraction and scattering often pose obstacles to light-sheet propagation and limit imaging depth. This is typically addressed by imaging multiple complementary views to obtain high and uniform image quality throughout the sample. However, multi-view imaging often requires complex multi-objective configurations that complicate sample mounting, or sample rotation that decreases imaging speed. Recent developments in single-objective light-sheet microscopy have shown that it is possible to achieve high spatio-temporal resolution with a single objective for both illumination and detection. Here we describe a single-objective light-sheet microscope that achieves: (i) high-resolution and large field-of-view imaging via a custom remote focusing objective; (ii) simpler design and ergonomics by remote placement of coverslips; (iii) fast volumetric imaging by means of light-sheet stabilised stage scanning – a novel scanning modality that extends the imaging volume without compromising imaging speed nor quality; (iv) multi-view imaging by means of dual orthogonal light-sheet illumination. Finally, we demonstrate the speed, field of view and resolution of our novel instrument by imaging zebrafish tail development.

Download Full-text

Aberration measurement and correction on a large field of view in fluorescence microscopy

Biomedical Optics Express ◽

10.1364/boe.441810 ◽

2021 ◽

Author(s):

Tommaso Furieri ◽

Daniele Ancora ◽

Gianmaria Calisesi ◽

stefano morara ◽

Andrea Bassi ◽

...

Keyword(s):

Fluorescence Microscopy ◽

Field Of View ◽

Large Field

Download Full-text

Resolution Enhancement in Phase Microscopy: a Review

10.20944/preprints201803.0273.v1 ◽

2018 ◽

Author(s):

Juanjuan Zheng ◽

Vicente Micó ◽

Peng Gao

Keyword(s):

Degrees Of Freedom ◽

Resolution Enhancement ◽

Field Of View ◽

Large Field ◽

Phase Imaging ◽

Structured Illumination ◽

Refractive Index Distribution ◽

Phase Microscopy ◽

Index Distribution ◽

Pros And Cons

Quantitative phase microscopy (QPM), a technique combining phase imaging and microscopy, enables visualization of the 3-D topography in reflective samples as well as the inner structure or refractive index distribution of transparent and translucent samples. However, as in conventional optical microscopy, QPM provides either a large field of view (FOV) or a high resolution but not both. Many approaches such as oblique illumination, structured illumination and speckle illumination have been proposed to improve the spatial resolution of phase microscopy by restricting other degrees of freedom (mostly time). Therefore, the space bandwidth product (SBP) of QPM becomes enlarged. This paper aims to provide an up-to-date review on the resolution enhancement approaches of QPM, discussing the pros and cons of each technique as well as the confusion on resolution definition claim on QPM and other coherent microscopy.

Download Full-text

Dual-phone illumination-imaging system for high resolution and large field of view multi-modal microscopy

Lab on a Chip ◽

10.1039/c8lc00995c ◽

2019 ◽

Vol 19 (5) ◽

pp. 825-836 ◽

Cited By ~ 5

Author(s):

Sara Kheireddine ◽

Ayyappasamy Sudalaiyadum Perumal ◽

Zachary J. Smith ◽

Dan V. Nicolau ◽

Sebastian Wachsmann-Hogiu

Keyword(s):

High Resolution ◽

Fluorescence Microscopy ◽

Mobile Phone ◽

Imaging System ◽

Dark Field ◽

Field Of View ◽

Large Field ◽

Bright Field

Bright-field, dark-field, Rheinberg, fluorescence microscopy on a mobile phone with phone screen illumination.

Download Full-text

Instrument concepts for the observation of prominences with future ground-based telescopes

Proceedings of the International Astronomical Union ◽

10.1017/s1743921313011216 ◽

2013 ◽

Vol 8 (S300) ◽

pp. 370-375 ◽

Cited By ~ 1

Author(s):

A. López Ariste

Keyword(s):

Image Quality ◽

Magnetic Fields ◽

Atmospheric Turbulence ◽

Temporal Resolution ◽

Field Of View ◽

Large Field ◽

Solar Observations ◽

New Generation

AbstractThe observation of prominences with ground-based telescopes suffers from poor image quality due to atmospheric turbulence when compared with space-borne instruments which, for solar observations, are of similar apertures. To make ground-based instruments competitive, they should rely on spectropolarimetry and the measurement of prominence magnetic fields, a task which no foreseable space instrument will perform. But spectropolarimetry alone does not suffice, and we argue that future instrumentation should combine it with imaging in a large field of view and good temporal resolution. We place numbers on those requirements and give examples of instrumental accomplishments already at work today that forecast a new generation of instruments for the observation of prominences from ground-based telescopes.

Download Full-text

Detection of cerebral tauopathy in P301L mice using high-resolution large-field multifocal illumination fluorescence microscopy

10.1101/2020.05.18.101188 ◽

2020 ◽

Author(s):

Ruiqing Ni ◽

Zhenyue Chen ◽

Juan A. Gerez ◽

Gloria Shi ◽

Quanyu Zhou ◽

...

Keyword(s):

High Resolution ◽

Fluorescence Microscopy ◽

Field Of View ◽

Large Field ◽

Depth Of Focus ◽

Microscopy Technique ◽

Microscopy Techniques ◽

Small Field Of View

AbstractCurrent intravital microscopy techniques visualize tauopathy with high-resolution, but have a small field-of-view and depth-of-focus. Herein, we report a transcranial detection of tauopathy over the entire cortex of P301L tauopathy mice using large-field multifocal illumination (LMI) fluorescence microscopy technique and luminescent conjugated oligothiophenes. In vitro assays revealed that fluorescent ligand h-FTAA is optimal for in vivo tau imaging, which was confirmed by observing elevated probe retention in the cortex of P301L mice compared to non-transgenic littermates. Immunohistochemical staining further verified the specificity of h-FTAA to detect tauopathy in P301L mice. The new imaging platform can be leveraged in pre-clinical mechanistic studies of tau spreading and clearance as well as longitudinal monitoring of tau targeting therapeutics.

Download Full-text