Automated imaging of cellular spheroids with selective plane illumination microscopy on a chip (Conference Presentation)

2017 ◽  
Author(s):  
Petra Paiè ◽  
Andrea Bassi ◽  
Francesca Bragheri ◽  
Roberto Osellame
Keyword(s):  
Cellular Spheroids ◽  
Automated Imaging ◽  
Selective Plane Illumination Microscopy

scholarly journals Live-Cell Imaging of Caspase Activation for High-Content Screening

2009 ◽  
Vol 14 (8) ◽  
pp. 956-969 ◽  
Author(s):  
Christophe Antczak ◽  
Toshimitsu Takagi ◽  
Christina N. Ramirez ◽  
Constantin Radu ◽  
Hakim Djaballah
Keyword(s):  
Cell Death ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Exposure Period ◽  
Live Cells ◽  
Assay Method ◽  
Caspase Activation ◽  
Fluorogenic Substrate ◽  
Automated Imaging ◽  
First Time

Caspases are central to the execution of programmed cell death, and their activation constitutes the biochemical hallmark of apoptosis. In this article, the authors report the successful adaptation of a high-content assay method using the DEVDNucView488™ fluorogenic substrate, and for the first time, they show caspase activation in live cells induced by either drugs or siRNA. The fluorogenic substrate was found to be nontoxic over an exposure period of several days, during which the authors demonstrate automated imaging and quantification of caspase activation of the same cell population as a function of time. Overexpression of the antiapoptotic protein Bcl-XL, alone or in combination with the inhibitor Z-VAD-FMK, attenuated caspase activation in HeLa cells exposed to doxorubicin, etoposide, or cell death siRNA. This method was further validated against 2 well-characterized NSCLC cell lines reported to be sensitive (H3255) or refractory (H2030) to erlotinib, where the authors show a differential time-dependent activation was observed for H3255 and no significant changes in H2030, consistent with their respective chemosensitivity profile. In summary, the results demonstrate the feasibility of using this newly adapted and validated high-content assay to screen chemical or RNAi libraries for the identification of previously uncovered enhancers and suppressors of the apoptotic machinery in live cells. ( Journal of Biomolecular Screening 2009:956-969)

scholarly journals Development of a Kinetic Assay for Late Endosome Movement

2014 ◽  
Vol 19 (7) ◽  
pp. 1070-1078 ◽  
Author(s):  
Milan Esner ◽  
Felix Meyenhofer ◽  
Michael Kuhn ◽  
Melissa Thomas ◽  
Yannis Kalaidzidis ◽  
...  
Keyword(s):  
Kinetic Parameters ◽  
Membrane Integrity ◽  
Living Cells ◽  
Mechanisms Of Action ◽  
Added Value ◽  
Kinetic Assay ◽  
Late Endosomes ◽  
Automated Imaging ◽  
Phenotypic Clustering ◽  
Large Compound

Automated imaging screens are performed mostly on fixed and stained samples to simplify the workflow and increase throughput. Some processes, such as the movement of cells and organelles or measuring membrane integrity and potential, can be measured only in living cells. Developing such assays to screen large compound or RNAi collections is challenging in many respects. Here, we develop a live-cell high-content assay for tracking endocytic organelles in medium throughput. We evaluate the added value of measuring kinetic parameters compared with measuring static parameters solely. We screened 2000 compounds in U-2 OS cells expressing Lamp1-GFP to label late endosomes. All hits have phenotypes in both static and kinetic parameters. However, we show that the kinetic parameters enable better discrimination of the mechanisms of action. Most of the compounds cause a decrease of motility of endosomes, but we identify several compounds that increase endosomal motility. In summary, we show that kinetic data help to better discriminate phenotypes and thereby obtain more subtle phenotypic clustering.

scholarly journals Label-free redox imaging of patient-derived organoids using selective plane illumination microscopy

2020 ◽  
Vol 11 (5) ◽  
pp. 2591 ◽  
Author(s):  
Peter F. Favreau ◽  
Jiaye He ◽  
Daniel A. Gil ◽  
Dustin A. Deming ◽  
Jan Huisken ◽  
...  
Keyword(s):  
Label Free ◽  
Selective Plane Illumination Microscopy

Selective Plane Illumination Microscopy (SPIM)

2013 ◽  
pp. 2307-2308
Author(s):  
Zeno Lavagnino ◽  
Francesca Cella Zanacchi ◽  
Alberto Diaspro
Keyword(s):  
Selective Plane Illumination Microscopy

Measuring the Confluence of iPSCs using an Automated Imaging System.

10.3791/61225 ◽  
2020 ◽  
Author(s):  
Valentina Magliocca ◽  
Maria Vinci ◽  
Tiziana Persichini ◽  
Franco Locatelli ◽  
Marco Tartaglia ◽  
...  
Keyword(s):  
Imaging System ◽  
Automated Imaging
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