Automated high-throughput light-sheet fluorescence microscopy of larval zebrafish

Logan, Savannah L.; Dudley, Christopher; Baker, Ryan; Taormina, Michael J.; Hay, Edouard A.; Parthasarathy, Raghuveer

doi:10.1371/journal.pone.0198705

Cited by 21 publications

(11 citation statements)

References 34 publications

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“… 127 This is an area that will grow in the coming years, as it is now being explored for drug discovery and precision medicine. 128 However, there are major limitations of 3D cell culture that still need to be overcome, namely, of: (i) scalability to multi-well microplates, (ii) automation, (iii) need of appropriate imaging systems for the visualization of 3D structures in an high throughput fashion (e.g., automated high-throughput light-sheet fluorescence microscopy that was only launched in 2018), 129 (iv) compatibility of systems (from NP library synthesis and liquid handling equipment to automated screening and analysis), and (v) reproducibility.…”

Section: Future Perspectivesmentioning

confidence: 99%

High-throughput screening of nanoparticles in drug delivery

et al. 2021

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The use of pharmacologically active compounds to manage and treat diseases is of utmost relevance in clinical practice. It is well recognized that spatial-temporal control over the delivery of these biomolecules will greatly impact their pharmacokinetic profile and ultimately their therapeutic effect. Nanoparticles (NPs) prepared from different materials have been tested successfully in the clinic for the delivery of several biomolecules including non-coding RNAs (siRNA and miRNA) and mRNAs. Indeed, the recent success of mRNA vaccines is in part due to progress in the delivery systems (NP based) that have been developed for many years. In most cases, the identification of the best formulation was done by testing a small number of novel formulations or by modification of pre-existing ones. Unfortunately, this is a low throughput and time-consuming process that hinders the identification of formulations with the highest potential. Alternatively, high-throughput combinatorial design of NP libraries may allow the rapid identification of formulations with the required release and cell/tissue targeting profile for a given application. Combinatorial approaches offer several advantages over conventional methods since they allow the incorporation of multiple components with varied chemical properties into materials, such as polymers or lipid-like materials, that will subsequently form NPs by self-assembly or chemical conjugation processes. The current review highlights the impact of high-throughput in the development of more efficient drug delivery systems with enhanced targeting and release kinetics. It also describes the current challenges in this research area as well as future directions.

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Section: Future Perspectivesmentioning

confidence: 99%

High-throughput screening of nanoparticles in drug delivery

et al. 2021

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“…This is especially important in the case of highly scattering samples such as spheroid, embryos, and larvae, in which it might be interesting to visualize specific structures or organs. In order to image fluorescent neutrophils in live zebrafish larvae, a fluidic circuit was used in which the samples were stopped in the detection field of view (FoV) 25 . Still, no rotation control of the specimen was implemented, and the use of glass capillaries would inevitably generate refractive index mismatches in both the illumination and detection paths, limiting also the resolution power.…”

Section: Introductionmentioning

confidence: 99%

Modular multimodal platform for classical and high throughput light sheet microscopy

Bernardello

Gualda

Loza-Álvarez

2022

Sci Rep

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Light-sheet fluorescence microscopy (LSFM) has become an important tool for biological and biomedical research. Although several illumination and detection strategies have been developed, the sample mounting still represents a cumbersome procedure as this is highly dependent on the type of sample and often this might be time consuming. This prevents the use of LSFM in other promising applications in which a fast and straightforward sample-mounting procedure and imaging are essential. These include the high-throughput research fields, e.g. in drug screenings and toxicology studies. Here we present a new imaging paradigm for LSFM, which exploits modularity to offer multimodal imaging and straightforward sample mounting strategy, enhancing the flexibility and throughput of the system. We describe its implementation in which the sample can be imaged either as in any classical configuration, as it flows through the light-sheet using a fluidic approach, or a combination of both. We also evaluate its ability to image a variety of samples, from zebrafish embryos and larvae to 3D complex cell cultures.

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“…Bright-field microscopy with a high-speed camera is the most common tool for measuring the blood velocity; however, the image contrast has remained low due to the transparent nature of red blood cells (RBCs). 5 , 6 Fluorescence microscopy has been introduced for a better image contrast with fluorescent labeling; 7 – 9 however, it requires a transgenic modified zebrafish for imaging, which is phototoxic and time-consuming.…”

Section: Introductionmentioning

confidence: 99%