Light-sheet fluorescent microscopy: fundamentals, developments and applications

Kafian, Hosein; Mozaffari-Jovin, Sina; Bagheri, Mohammad; Mousavi Shaegh, Seyed Ali

doi:10.1088/1402-4896/acd7ae

Cited by 4 publications

(3 citation statements)

References 156 publications

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“…Moreover, some organic molecules in biological samples can be decomposed by long-term laser point-by-point scanning, causing phototoxicity. These drawbacks can be addressed using light sheet fluorescence microscopy (discussed hereafter), which provides high spatio-temporal resolution while measuring samples within the dimension from submicron to centimeters in size [ 65 ].…”

Section: Optical Imaging Detection Techniquesmentioning

confidence: 99%

On-Chip Photonic Detection Techniques for Non-Invasive In Situ Characterizations at the Microfluidic Scale

Kurdadze,

Lamadie,

Nehme

et al. 2024

Sensors

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Microfluidics has emerged as a robust technology for diverse applications, ranging from bio-medical diagnostics to chemical analysis. Among the different characterization techniques that can be used to analyze samples at the microfluidic scale, the coupling of photonic detection techniques and on-chip configurations is particularly advantageous due to its non-invasive nature, which permits sensitive, real-time, high throughput, and rapid analyses, taking advantage of the microfluidic special environments and reduced sample volumes. Putting a special emphasis on integrated detection schemes, this review article explores the most relevant advances in the on-chip implementation of UV–vis, near-infrared, terahertz, and X-ray-based techniques for different characterizations, ranging from punctual spectroscopic or scattering-based measurements to different types of mapping/imaging. The principles of the techniques and their interest are discussed through their application to different systems.

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Section: Optical Imaging Detection Techniquesmentioning

confidence: 99%

On-Chip Photonic Detection Techniques for Non-Invasive In Situ Characterizations at the Microfluidic Scale

Kurdadze,

Lamadie,

Nehme

et al. 2024

Sensors

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“…Here, we will demonstrate the potential of DD in the context of Light-Sheet Fluorescence Microscopy (LSFM), which necessitates the incorporation of a very complex optical model into the optimization process. LSFM has become an indispensable imaging tool in the life science community due to its high acquisition speed and optical sectioning capabilities [26][27][28][29][30][31]. In LSFM, the illumination path is separated and orthogonal to the detection path.…”

Section: Introductionmentioning

confidence: 99%

“…LSFM image quality depends on various factors. Lateral resolution is influenced by fluorescence wavelength and detection objective numerical aperture (NA), while axial resolution and signalto-noise ratio are broadly influenced by the illumination pattern shape and detection objective NA [31,34]. As a result, the image quality of LSFM is not uniform across the Field of View (FOV), often resulting in less-than-ideal datasets.…”

Section: Introductionmentioning

confidence: 99%

Enhancing Light-Sheet Fluorescence Microscopy Illumination Beams through Deep Design Optimization

Li,

Rai,

Cai

et al. 2023

Preprint

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Light sheet fluorescence microscopy (LSFM) provides the benefit of optical sectioning coupled with rapid acquisition times for imaging of tissue-cleared specimen. This allows for high-resolution 3D imaging of large tissue volumes. Inherently to LSFM, the quality of the imaging heavily relies on the characteristics of the illumination beam, with the notion that the illumination beam only illuminates a thin section that is being imaged. Therefore, substantial efforts are dedicated to identifying slender, non-diffracting beam profiles that can yield uniform and high-contrast images. An ongoing debate concerns the employment of the most optimal illumination beam; Gaussian, Bessel, Airy patterns and/or others. Comparisons among different beam profiles is challenging as their optimization objective is often different. Given that our large imaging datasets (∼0.5TB images per sample) is already analyzed using deep learning models, we envisioned a different approach to this problem by hypothesizing that we can tailor the illumination beam to boost the deep learning models performance. We achieve this by integrating the physical LSFM illumination model after passing through a variable phase mask into the training of a cell detection network. Here we report that the joint optimization continuously updates the phase mask, improving the image quality for better cell detection. Our method’s efficacy is demonstrated through both simulations and experiments, revealing substantial enhancements in imaging quality compared to traditional Gaussian light sheet. We offer valuable insights for designing microscopy systems through a computational approach that exhibits significant potential for advancing optics design that relies on deep learning models for analysis of imaging datasets.

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Expansion and Light‐Sheet Microscopy for Nanoscale 3D Imaging

Pesce,

Ricci,

Sportelli

et al. 2024

Small Methods

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Expansion Microscopy (ExM) and Light‐Sheet Fluorescence Microscopy (LSFM) are forefront imaging techniques that enable high‐resolution visualization of biological specimens. ExM enhances nanoscale investigation using conventional fluorescence microscopes, while LSFM offers rapid, minimally invasive imaging over large volumes. This review explores the joint advancements of ExM and LSFM, focusing on the excellent performance of the integrated modality obtained from the combination of the two, which is refer to as ExLSFM. In doing so, the chemical processes required for ExM, the tailored optical setup of LSFM for examining expanded samples, and the adjustments in sample preparation for accurate data collection are emphasized. It is delve into various specimen types studied using this integrated method and assess its potential for future applications. The goal of this literature review is to enrich the comprehension of ExM and LSFM, encouraging their wider use and ongoing development, looking forward to the upcoming challenges, and anticipating innovations in these imaging techniques.

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Light-sheet fluorescent microscopy: fundamentals, developments and applications

Cited by 4 publications

References 156 publications

On-Chip Photonic Detection Techniques for Non-Invasive In Situ Characterizations at the Microfluidic Scale

On-Chip Photonic Detection Techniques for Non-Invasive In Situ Characterizations at the Microfluidic Scale

Enhancing Light-Sheet Fluorescence Microscopy Illumination Beams through Deep Design Optimization

Expansion and Light‐Sheet Microscopy for Nanoscale 3D Imaging

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