Abstract
Acquiring high-content images of 3D-cultured cells for analyzing multiple cellular events is a daunting task, requiring an automated fluorescent microscope and high-throughput image analysis software. High throughput is an important feature for high-content imaging (HCI) devices to enable rapid image acquisition. Various factors come into play when dealing with the speed of image acquisition. For example, capturing large number of cells with lower magnification or reducing sample volume/size can increase data acquisition speed. In addition, reducing exposure time with the use of high intensity light sources, optimizing fluorescence staining protocols for brighter colors, and using an objective lens with relatively high numerical aperture also significantly increases the image acquisition speed [1].
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References
Buchser, W., Collins, M., Garyantes, T., Guha, R., Haney, S., Lemmon, V., et al. (2012). Assay development guidelines for image-based high content screening, high content analysis and high content imaging. In S. GS, C. NP, H. Nelson, M. Arkin, D. Auld, C. Austin, et al. (Eds.), Assay guidance manual. (pp. 1–69). Bethesda, MD: Eli Lilly & Company and the National Center for Advancing Translational Sciences.
Talbot, C. B., McGinty, J., Grant, D. M., McGhee, E. J., Owen, D. M., Zhang, W., et al. (2008). High speed unsupervised fluorescence lifetime imaging confocal multiwell plate reader for high content analysis. Journal of Biophotonics, 1(6), 514–521. doi:10.1002/jbio.200810054.
Celli, J. P., Rizvi, I., Blanden, A. R., Massodi, I., Gidden, M. D., Pogue, B. W., et al. (2014). An imaging-based platform for high-content, quantitative evaluation of therapeutic response in 3D tumour models. Scientific Reports. doi:10.1038/srep03751.
Romano, S. N., & Gorelick, D. A. (2014). Semi-automated imaging of tissue-specific fluorescence in Zebrafish Embryos. Journal of Visualized Experiments, 87, e51533. doi:10.3791/51533.
Bickle, M. (2010). The beautiful cell: High-content screening in drug discovery. Analytical and Bioanalytical Chemistry, 398, 219–226. doi:10.1007/s00216-010-3788-3.
Jahr, W., Schmid, B., Schmied, C., Fahrbach, F. O., & Huisken, J. (2015). Hyperspectral light sheet microscopy. Nature Communications, 6, 1–7. doi:10.1038/ncomms8990.
Reynaud, E. G., Peychl, J., Huisken, J., & Tomancak, P. (2015). Guide to light-sheet microscopy for adventurous biologists. Nature Methods, 12(1), 30–34. doi:10.1038/nmeth.3222.
Rae Chi, K. (2012). High on high content: A guide to some new and improved high-content screening systems. The Scientist Magazine, 1–7.
Starkuviene, V., & Pepperkok, R. (2007). The potential of high-content high-throughput microscopy in drug discovery. British Journal of Pharmacology, 152(1), 62–71. doi:10.1038/sj.bjp.0707346.
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In the S+ Scanner, the following four filter sets are installed.
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Multiband filter
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Joshi, P., Yu, KN., Serbinowski, E., Lee, MY. (2016). 3D-Cultured Cell Image Acquisition. In: Lee, MY. (eds) Microarray Bioprinting Technology. Springer, Cham. https://doi.org/10.1007/978-3-319-46805-1_6
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DOI: https://doi.org/10.1007/978-3-319-46805-1_6
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