Abstract
High Content Imaging (HCI) is a quantitative automated fluorescence microscopy approach which uses advanced algorithms for rapid analysis of images acquired from fluorescently stained cells. The term “High Content” derives from the fact that HCI allows for multiplexing of various readouts by use of several fluorescence probes at different wavelengths. This method facilitates information generation on multiple simultaneous readouts, for instance on morphological structures, spatial and dynamic processes of molecules, signal transduction, or enzyme activities. Moreover, mechanisms of compound-induced toxicity and the specific cellular pathways involved may be studied by combining the HCI approach with the use of specific enzyme inhibitors, enzyme inducers or RNA interference. As HCI provides multiplexed detailed information at the level of a single cell, as well as characterization of cellular population distributions, it offers a superior investigational tool compared to standard spectrophotometric plate readers that measure only average properties of a cell population.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Taylor DL (2010) A personal perspective on high-content screening (HCS): from the beginning. J Biomol Screen 15(7):720–725
Ott M, Gogvadze V, Orrenius S, Zhivotovsky B (2007) Mitochondria, oxidative stress and cell death. Apoptosis 12(5):913–922
Stevenson D, Wokosin D, Girkin J, Grant MH (2002) Measurement of the intracellular distribution of reduced glutathione in cultured rat hepatocytes using monochlorobimane and confocal laser scanning microscopy. Toxicol In Vitro 16(5):609–619
Gutscher M, Pauleau AL, Marty L et al (2008) Real-time imaging of the intracellular glutathione redox potential. Nat Methods 5(6): 553–559
Lee HC, Yin PH, Lu CY, Chi CW, Wei YH (2000) Increase of mitochondria and mitochondrial DNA in response to oxidative stress in human cells. Biochem J 348(Pt 2):425–432
Kluza J, Marchetti P, Gallego MA et al (2004) Mitochondrial proliferation during apoptosis induced by anticancer agents: effects of doxorubicin and mitoxantrone on cancer and cardiac cells. Oncogene 23(42):7018–7030
Pendergrass W, Wolf N, Poot M (2004) Efficacy of MitoTracker Green and CMXrosamine to measure changes in mitochondrial membrane potentials in living cells and tissues. Cytometry A 61(2):162–169
Petrovas C, Mueller YM, Dimitriou ID et al (2007) Increased mitochondrial mass characterizes the survival defect of HIV-specific CD8(+) T cells. Blood 109(6):2505–2513
Perry SW, Norman JP, Barbieri J, Brown EB, Gelbard HA (2011) Mitochondrial membrane potential probes and the proton gradient: a practical usage guide. Biotechniques 50(2): 98–115
Martin SJ, Reutelingsperger CP, McGahon AJ et al (1995) Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med 182(5):1545–1556
Begriche K, Massart J, Robin MA, Borgne-Sanchez A, Fromenty B (2011) Drug-induced toxicity on mitochondria and lipid metabolism: mechanistic diversity and deleterious consequences for the liver. J Hepatol 54(4): 773–794
Labbe G, Pessayre D, Fromenty B (2008) Drug-induced liver injury through mitochondrial dysfunction: mechanisms and detection during preclinical safety studies. Fundam Clin Pharmacol 22(4):335–353
Halliwell WH (1997) Cationic amphiphilic drug-induced phospholipidosis. Toxicol Pathol 25(1):53–60
Morelli JK, Buehrle M, Pognan F et al (2006) Validation of an in vitro screen for phospholipidosis using a high-content biology platform. Cell Biol Toxicol 22(1):15–27
Borst P, Zelcer N, van de Wetering K (2006) MRP2 and 3 in health and disease. Cancer Lett 234(1):51–61
Funk C, Ponelle C, Scheuermann G, Pantze M (2001) Cholestatic potential of troglitazone as a possible factor contributing to troglitazone-induced hepatotoxicity: in vivo and in vitro interaction at the canalicular bile salt export pump (Bsep) in the rat. Mol Pharmacol 59(3): 627–635
Kamimoto Y, Gatmaitan Z, Hsu J, Arias IM (1989) The function of Gp170, the multidrug resistance gene product, in rat liver canalicular membrane vesicles. J Biol Chem 264(20): 11693–11698
Zamek-Gliszczynski MJ, Xiong H, Patel NJ et al (2003) Pharmacokinetics of 5 (and 6)-carboxy-2′,7′-dichlorofluorescein and its diacetate promoiety in the liver. J Pharmacol Exp Ther 304(2):801–809
Wilton JC, Coleman R, Lankester DJ, Chipman JK (1993) Stability and optimization of canalicular function in hepatocyte couplets. Cell Biochem Funct 11(3):179–185
Konya A, Andor A, Satorhelyi P, Nemeth K, Kurucz I (2006) Inhibition of the MDR1 transporter by new phenothiazine derivatives. Biochem Biophys Res Commun 346(1): 45–50
Meriane M, Tcherkezian J, Webber CA et al (2004) Phosphorylation of DCC by Fyn mediates Netrin-1 signaling in growth cone guidance. J Cell Biol 167(4):687–698
Monroy-Contreras R, Vaca L (2011) Molecular beacons: powerful tools for imaging RNA in living cells. J Nucleic Acids 2011: 741723
Rosenthal EL, Kulbersh BD, King T, Chaudhuri TR, Zinn KR (2007) Use of fluorescent labeled anti-epidermal growth factor receptor antibody to image head and neck squamous cell carcinoma xenografts. Mol Cancer Ther 6(4):1230–1238
O’Brien PJ, Irwin W, Diaz D et al (2006) High concordance of drug-induced human hepatotoxicity with in vitro cytotoxicity measured in a novel cell-based model using high content screening. Arch Toxicol 80(9): 580–604
Wolf A, Mueller SO, Hewitt P, Pfaller W, Jennings P, Testai E, Bois F, Prieto P, Price A, Lukas A, Richert L, Guillouzo A, Leonard M, Blaauboer BJ, Rosrami A, Reinert K, Honegger P, Cecchelli R, Kopp-Schneider A, Weiss DG, Schroeder O, Huber C, Dekant W (2013) Predict-IV project overview (EU grant 202222): non-animal based toxicity profiling by integrating toxicodynamics and biokinetics. Toxicol Lett 221S, S7
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Uteng, M., Germano, D., Balavenkatraman, K.K., Pognan, F., Wolf, A. (2014). High Content Imaging Approaches for In Vitro Toxicology. In: Bal-Price, A., Jennings, P. (eds) In Vitro Toxicology Systems. Methods in Pharmacology and Toxicology. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0521-8_17
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0521-8_17
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0520-1
Online ISBN: 978-1-4939-0521-8
eBook Packages: Springer Protocols