Confocal-Based Method for Quantification of Diffusion Kinetics in Microwell Plates and Its Application for Identifying a Rapid Mixing Method for High-Content/Throughput Screening

Song, Ok Ryul; Kim, Tae Hee; Perrodon, Xavier; Lee, Changbok; Jeon, Hee Kyoung; Seghiri, Zahir; Kwon, Ho Jeong; Cechetto, Jonathan; Christophe, Thierry

doi:10.1177/1087057109357787

Cited by 2 publications

(2 citation statements)

References 7 publications

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“…To ensure this, homogeneous exposure of the cells to chemicals during treatment is essential and must consider partitioning effects and diffusion kinetics within the medium. Adhering to recommended practices, such as those by Song et al ( Song et al, 2010 ), and integrating liquid handling systems in the procedure can support this. Nonetheless, comprehensive standard operating procedures for chemical and genetic perturbation of cells are still elusive and remain a challenge due to diverse technical equipment and protocols.…”

Section: Potential Limitations and Challenges Of Working With Cp Datamentioning

confidence: 95%

Unleashing the potential of cell painting assays for compound activities and hazards prediction

Odje,

Meijer,

von Coburg

et al. 2024

Front. Toxicol.

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The cell painting (CP) assay has emerged as a potent imaging-based high-throughput phenotypic profiling (HTPP) tool that provides comprehensive input data for in silico prediction of compound activities and potential hazards in drug discovery and toxicology. CP enables the rapid, multiplexed investigation of various molecular mechanisms for thousands of compounds at the single-cell level. The resulting large volumes of image data provide great opportunities but also pose challenges to image and data analysis routines as well as property prediction models. This review addresses the integration of CP-based phenotypic data together with or in substitute of structural information from compounds into machine (ML) and deep learning (DL) models to predict compound activities for various human-relevant disease endpoints and to identify the underlying modes-of-action (MoA) while avoiding unnecessary animal testing. The successful application of CP in combination with powerful ML/DL models promises further advances in understanding compound responses of cells guiding therapeutic development and risk assessment. Therefore, this review highlights the importance of unlocking the potential of CP assays when combined with molecular fingerprints for compound evaluation and discusses the current challenges that are associated with this approach.

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Section: Potential Limitations and Challenges Of Working With Cp Datamentioning

confidence: 95%

Unleashing the potential of cell painting assays for compound activities and hazards prediction

Odje,

Meijer,

von Coburg

et al. 2024

Front. Toxicol.

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“…To achieve this, assay miniaturization in the micro-titer plates and manipulation of library of molecules are needed to optimize the process of conservation, dispensing and mixing 13 . Furthermore, it is recommended to use an automated robotic platform, including dispensers, in order to accurately and reproducibly control assay conditions in the micro-titer plates.…”

Section: Discussionmentioning

confidence: 99%

A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening

Queval

Song

Delorme

et al. 2014

JoVE

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Despite the availability of therapy and vaccine, tuberculosis (TB) remains one of the most deadly and widespread bacterial infections in the world. Since several decades, the sudden burst of multi-and extensively-drug resistant strains is a serious threat for the control of tuberculosis. Therefore, it is essential to identify new targets and pathways critical for the causative agent of the tuberculosis, Mycobacterium tuberculosis (Mtb) and to search for novel chemicals that could become TB drugs. One approach is to set up methods suitable for the genetic and chemical screens of large scale libraries enabling the search of a needle in a haystack. To this end, we developed a phenotypic assay relying on the detection of fluorescently labeled Mtb within fluorescently labeled host cells using automated confocal microscopy. This in vitro assay allows an image based quantification of the colonization process of Mtb into the host and was optimized for the 384-well microplate format, which is proper for screens of siRNA-, chemical compound-or Mtb mutant-libraries. The images are then processed for multiparametric analysis, which provides read out inferring on the pathogenesis of Mtb within host cells. Video LinkThe video component of this article can be found at

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Confocal-Based Method for Quantification of Diffusion Kinetics in Microwell Plates and Its Application for Identifying a Rapid Mixing Method for High-Content/Throughput Screening

Cited by 2 publications

References 7 publications

Unleashing the potential of cell painting assays for compound activities and hazards prediction

Unleashing the potential of cell painting assays for compound activities and hazards prediction

A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening

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