{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T13:35:57Z","timestamp":1787319357570,"version":"build-2736575974"},"reference-count":44,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,12,6]],"date-time":"2025-12-06T00:00:00Z","timestamp":1764979200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T00:00:00Z","timestamp":1767744000000},"content-version":"vor","delay-in-days":32,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Determining whether potential cancer therapies effectively kill cancer cells is important for informing effective therapeutic choice for patients. Here we describe a rapid label-free method for testing drug efficacy in vitro that evaluates cellular viability from sub-cellular fluctuation imaging (SCFI). We used staurosporine and paclitaxel as known cytotoxic drugs at different concentrations, and four different human cancer-derived cell lines: PC3 (prostate), Caco-2 (colorectal), Calu-3 (lung) and A549 (lung). Both drugs caused a rapid decrease in sub-cellular fluctuations within 1 to 3 h except when the specific cell line was known to be resistant to one of the drugs. We also demonstrated that the method is able to differentiate between treated and untreated PC3 cells within 3 to 4 h after cells have been plated, thus eliminating the need for overnight incubation, and further decreasing the total time needed to evaluate drug efficacy. SCFI is therefore able to identify reliably if drugs are cytotoxic within 3 h of addition, which is considerably faster than current commonly used techniques.<\/jats:p>","DOI":"10.1038\/s41598-025-30295-9","type":"journal-article","created":{"date-parts":[[2025,12,6]],"date-time":"2025-12-06T12:25:05Z","timestamp":1765023905000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A rapid method of evaluating cytotoxic drug efficacy using sub-cellular fluctuation imaging"],"prefix":"10.1038","volume":"16","author":[{"given":"Henrik","family":"Rehnstrom","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arthur","family":"Eley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natasha S.","family":"Clayton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georgina","family":"Plant","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diego","family":"Oviedo-Chavez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ben","family":"Ede","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Darryl J.","family":"Hill","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne J.","family":"Ridley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Massimo","family":"Antognozzi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,6]]},"reference":[{"key":"30295_CR1","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1038\/483531a","volume":"483","author":"CG Begley","year":"2012","unstructured":"Begley, C. G. & Ellis, L. M. Raise standards for preclinical cancer research. Nature 483, 531\u2013533 (2012).","journal-title":"Nature"},{"key":"30295_CR2","doi-asserted-by":"publisher","first-page":"1324","DOI":"10.2174\/0929867322666150209150639","volume":"22","author":"M Lopez-Lazaro","year":"2015","unstructured":"Lopez-Lazaro, M. A simple and reliable approach for assessing anticancer activity in vitro. Curr. Med. Chem. 22, 1324\u20131334 (2015).","journal-title":"Curr. Med. Chem."},{"key":"30295_CR3","doi-asserted-by":"publisher","first-page":"33","DOI":"10.2174\/1875397300903010033","volume":"3","author":"AL Niles","year":"2009","unstructured":"Niles, A. L., Moravec, R. A. & Riss, T. L. In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high throughput screening. Curr. Chem. Genomics 3, 33\u201341 (2009).","journal-title":"Curr. Chem. Genomics"},{"key":"30295_CR4","doi-asserted-by":"publisher","first-page":"171","DOI":"10.1016\/j.toxlet.2005.07.001","volume":"160","author":"G Fotakis","year":"2005","unstructured":"Fotakis, G. & Timbrell, J. A. In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol. Lett. 160, 171\u2013177 (2005).","journal-title":"Toxicol. Lett."},{"key":"30295_CR5","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1016\/0022-1759(95)00072-I","volume":"184","author":"I Vermes","year":"1995","unstructured":"Vermes, I. et al. A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J. Immunol. Methods 184, 39\u201351 (1995).","journal-title":"J. Immunol. Methods"},{"key":"30295_CR6","doi-asserted-by":"publisher","first-page":"168","DOI":"10.1016\/0003-2697(88)90532-5","volume":"174","author":"Y Kim","year":"1988","unstructured":"Kim, Y. et al. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal. Biochem. 174, 168\u2013176 (1988).","journal-title":"Anal. Biochem."},{"key":"30295_CR7","doi-asserted-by":"publisher","first-page":"4835","DOI":"10.1073\/pnas.74.11.4835","volume":"74","author":"G McMaster","year":"1977","unstructured":"McMaster, G. & Carmichael, G. Analysis of single and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. PNAS 74, 4835\u20134838 (1977).","journal-title":"PNAS"},{"key":"30295_CR8","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1016\/0022-1759(91)90198-O","volume":"193","author":"I Nicoletti","year":"1991","unstructured":"Nicoletti, I. et al. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J. Immunol. Methods 193, 271\u2013279 (1991).","journal-title":"J. Immunol. Methods"},{"key":"30295_CR9","doi-asserted-by":"publisher","first-page":"4016","DOI":"10.1158\/0008-5472.CAN-14-0725","volume":"74","author":"C Hatzis","year":"2014","unstructured":"Hatzis, C. et al. Enhancing reproducibility in cancer drug screening: How do we move forward?. Cancer Res. 74, 4016\u20134023 (2014).","journal-title":"Cancer Res."},{"key":"30295_CR10","doi-asserted-by":"publisher","first-page":"1147","DOI":"10.1126\/science.1097640","volume":"305","author":"AE Pelling","year":"2004","unstructured":"Pelling, A. E., Sehati, S., Gralla, E. B., Valentine, E. S. & Gimzewski, J. K. Local nanomechanical motion of the cell wall of Saccharomyces cerevisiae. Science 305, 1147\u20131150 (2004).","journal-title":"Science"},{"key":"30295_CR11","doi-asserted-by":"publisher","first-page":"378","DOI":"10.1073\/pnas.1415348112","volume":"112","author":"S Kasas","year":"2015","unstructured":"Kasas, S. et al. Detecting nanoscale vibrations as signature of life. Proc. Natl. Acad. Sci. 112, 378\u2013381 (2015).","journal-title":"Proc. Natl. Acad. Sci."},{"key":"30295_CR12","doi-asserted-by":"publisher","first-page":"522","DOI":"10.1038\/nnano.2013.120","volume":"8","author":"G Longo","year":"2013","unstructured":"Longo, G. et al. Rapid detection of bacterial resistance to antibiotics using AFM cantilevers as nanomechanical sensors. Nat. Nanotechnol. 8, 522\u2013526 (2013).","journal-title":"Nat. Nanotechnol."},{"key":"30295_CR13","doi-asserted-by":"publisher","first-page":"590","DOI":"10.1002\/jmr.2306","volume":"26","author":"S Aghayee","year":"2013","unstructured":"Aghayee, S. et al. Combination of fluorescence microscopy and nanomotion detection to characterize bacteria. J. Mol. Recognit. 26, 590\u2013595 (2013).","journal-title":"J. Mol. Recognit."},{"key":"30295_CR14","doi-asserted-by":"publisher","first-page":"113701","DOI":"10.1063\/1.4895132","volume":"105","author":"C Lissandrello","year":"2014","unstructured":"Lissandrello, C. et al. Nanomechanical motion of Escherichia coli adhered to a surface. Appl. Phys. Lett. 105, 113701 (2014).","journal-title":"Appl. Phys. Lett."},{"key":"30295_CR15","doi-asserted-by":"publisher","first-page":"12947","DOI":"10.1038\/ncomms12947","volume":"7","author":"H Etayahi","year":"2016","unstructured":"Etayahi, H., Khan, M. F., Kaur, K. & Thundat, T. Microfluidic cantilever detects bacteria and measures their susceptibility to antibiotics in small confined volumes. Nat. Commun. 7, 12947 (2016).","journal-title":"Nat. Commun."},{"key":"30295_CR16","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/j.bios.2014.06.069","volume":"63","author":"K Syal","year":"2015","unstructured":"Syal, K. et al. Plasmonic imaging of protein interactions with single bacterial cells. Biosens. Bioelectron. 63, 131\u2013137 (2015).","journal-title":"Biosens. Bioelectron."},{"key":"30295_CR17","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1021\/acsnano.5b05944","volume":"10","author":"K Syal","year":"2015","unstructured":"Syal, K. et al. Antimicrobial susceptibility test with plasmonic imaging and tracking of single bacterial motions on nanometer scale. ACS Nano 10, 845\u2013852 (2015).","journal-title":"ACS Nano"},{"key":"30295_CR18","doi-asserted-by":"publisher","first-page":"743","DOI":"10.1039\/C7LC01019B","volume":"18","author":"V Kara","year":"2018","unstructured":"Kara, V. et al. Microfluidic detection of movements of Escherichia coli for rapid antibiotic susceptibility testing. Lab Chip 18, 743\u2013753 (2018).","journal-title":"Lab Chip"},{"key":"30295_CR19","doi-asserted-by":"publisher","first-page":"12138","DOI":"10.1038\/s41598-017-12063-6","volume":"7","author":"WL Johnson","year":"2017","unstructured":"Johnson, W. L., France, D. C., Rentz, N. S., Cordell, W. T. & Walls, F. L. Sensing bacterial vibrations and early response to antibiotics with phase noise of a resonant crystal. Sci. Rep. 7, 12138 (2017).","journal-title":"Sci. Rep."},{"key":"30295_CR20","doi-asserted-by":"publisher","first-page":"1283","DOI":"10.1007\/s00253-019-10295-2","volume":"104","author":"OI Guliy","year":"2020","unstructured":"Guliy, O. I., Zaitsev, B. D. & Borodina, I. A. New approach for determination of antimicrobial susceptibility to antibiotics by an acoustic sensor. Appl. Microbiol. Biotechnol. 104, 1283\u20131290 (2020).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"30295_CR21","doi-asserted-by":"publisher","unstructured":"Bermingham, C. R. et al. Imaging of sub-cellular fluctuations provides a rapid way to observe bacterial viability and response to antibiotics. Sensors (preprint at bioRxiv.org). TBC. https:\/\/doi.org\/10.1101\/460139 (2025).","DOI":"10.1101\/460139"},{"key":"30295_CR22","doi-asserted-by":"publisher","first-page":"1749","DOI":"10.1086\/647952","volume":"11","author":"JH Jorgensen","year":"2009","unstructured":"Jorgensen, J. H. & Ferraro, M. J. Antimicrobial susceptibility testing - A review of general principles and contemporary practices. Clin. Infect. Dis. 11, 1749\u20131755 (2009).","journal-title":"Clin. Infect. Dis."},{"key":"30295_CR23","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1016\/j.drup.2011.02.002","volume":"14","author":"IN Okeke","year":"2011","unstructured":"Okeke, I. N. et al. Diagnostics as essential tools for containing antibacterial resistance. Drug Resist. Updates 14, 95\u2013106 (2011).","journal-title":"Drug Resist. Updates"},{"key":"30295_CR24","doi-asserted-by":"publisher","first-page":"1511","DOI":"10.1364\/BOE.7.001511","volume":"7","author":"C Apelian","year":"2016","unstructured":"Apelian, C. et al. Dynamic full field optical coherence tomography: subcellular metabolic contrast revealed in tissues by interferometric signals temporal analysis. Biomed. Opt. Exp. 7, 1511\u20131524 (2016).","journal-title":"Biomed. Opt. Exp."},{"key":"30295_CR25","doi-asserted-by":"publisher","first-page":"384002","DOI":"10.1088\/0957-4484\/19\/38\/384002","volume":"19","author":"M Antognozzi","year":"2008","unstructured":"Antognozzi, M. et al. A new detection system for extremely small vertically mounted cantilevers. Nanotechnology 19, 384002 (2008).","journal-title":"Nanotechnology"},{"key":"30295_CR26","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1038\/nphys3732","volume":"12","author":"M Antognozzi","year":"2016","unstructured":"Antognozzi, M. et al. Direct measurements of the extraordinary optical momentum and traverse spin-dependent force using a nano-cantilever. Nat. Phys. 12, 731\u2013735 (2016).","journal-title":"Nat. Phys."},{"key":"30295_CR27","doi-asserted-by":"publisher","first-page":"1948","DOI":"10.1038\/nprot.2015.100","volume":"10","author":"JW Krieger","year":"2015","unstructured":"Krieger, J. W. et al. Imaging fluorescence (cross-) correlation spectroscopy in live cells and organisms. Nat. Protoc. 10, 1948\u20131974 (2015).","journal-title":"Nat. Protoc."},{"key":"30295_CR28","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1007\/s12013-007-9000-5","volume":"49","author":"D Kolin","year":"2007","unstructured":"Kolin, D. & Wiseman, P. Advances in image correlation spectroscopy: Measuring number densities, aggregation states, and dynamics of fluorescently labeled macromolecules in cells. Cell Biochem. Biophys. 49, 141\u2013164 (2007).","journal-title":"Cell Biochem. Biophys."},{"key":"30295_CR29","doi-asserted-by":"publisher","first-page":"336","DOI":"10.1101\/pdb.top086124","volume":"2015","author":"PW Wiseman","year":"2015","unstructured":"Wiseman, P. W. Image correlation spectroscopy: Principles and applications. Cold Spring Harbor Protoc. 2015, 336\u2013348 (2015).","journal-title":"Cold Spring Harbor Protoc."},{"key":"30295_CR30","first-page":"1699","volume":"61","author":"XY Li","year":"2001","unstructured":"Li, X. Y. et al. Overexpression of BCL-X-L underlies the molecular basis for resistance to staurosporine-induced apoptosis in PC-3 cells. Cancer Res. 61, 1699\u20131706 (2001).","journal-title":"Cancer Res."},{"key":"30295_CR31","doi-asserted-by":"publisher","first-page":"20301","DOI":"10.1074\/jbc.M201604200","volume":"23","author":"LK Nutt","year":"2002","unstructured":"Nutt, L. K. et al. Bax-mediated Ca2+ mobilization promotes cytochrome c release during apoptosis. J. Biol. Chem. 23, 20301\u201320308 (2002).","journal-title":"J. Biol. Chem."},{"key":"30295_CR32","doi-asserted-by":"publisher","first-page":"1317","DOI":"10.1158\/1078-0432.CCR-05-1634","volume":"12","author":"N Desai","year":"2006","unstructured":"Desai, N. et al. Increased antitumoractivity, intratumor paclitaxel concentrations, and endothelial cell transport of cremophor-free, albumin-bound paclitaxel, ABI-007, compared with cremophor-based paclitaxel. Clin. Cancer Res. 12, 1317\u20131324 (2006).","journal-title":"Clin. Cancer Res."},{"key":"30295_CR33","doi-asserted-by":"publisher","first-page":"755","DOI":"10.1517\/14656566.3.6.755","volume":"3","author":"T Mekhail","year":"2002","unstructured":"Mekhail, T. & Markman, M. Paclitaxel in cancer therapy. Expert Opin. Pharmacother. 3, 755\u2013766 (2002).","journal-title":"Expert Opin. Pharmacother."},{"key":"30295_CR34","doi-asserted-by":"publisher","first-page":"1454","DOI":"10.1002\/(SICI)1097-0142(19990401)85:7<1454::AID-CNCR5>3.0.CO;2-#","volume":"85","author":"BC Sheppard","year":"1999","unstructured":"Sheppard, B. C. et al. Effects of paclitaxel on the growth of normal, polyposis, and cancerous human colonic epithelial cells. Cancer 85, 1454\u20131464 (1999).","journal-title":"Cancer"},{"key":"30295_CR35","first-page":"465","volume":"267","author":"M Bissonnette","year":"1994","unstructured":"Bissonnette, M. et al. 1.25(OH)2 vitamin D3 activates PKC-$$\\alpha$$ in Caco-2 cells. Am. J. Physiol. 267, 465\u2013475 (1994).","journal-title":"Am. J. Physiol."},{"key":"30295_CR36","doi-asserted-by":"publisher","first-page":"1025","DOI":"10.1006\/cbir.1995.1045","volume":"19","author":"MD Basson","year":"1995","unstructured":"Basson, M. D. & Hong, F. Modulation of human Caco-2 intestinal epithelian cell phenotype by protein kinase C inhibitors. Cell Biol. Int. 19, 1025\u20131032 (1995).","journal-title":"Cell Biol. Int."},{"key":"30295_CR37","doi-asserted-by":"publisher","first-page":"4260","DOI":"10.1128\/IAI.71.8.4260-4270.2003","volume":"71","author":"G Chakrabarti","year":"2003","unstructured":"Chakrabarti, G., Zhou, X. & McClane, B. A. Death pathways activated in Caco-2 cells by Clostridium perfringens enterotoxin. Infect. Immun. 71, 4260\u20134270 (2003).","journal-title":"Infect. Immun."},{"key":"30295_CR38","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1016\/0006-291X(86)90008-2","volume":"135","author":"T Tamaoki","year":"1986","unstructured":"Tamaoki, T. et al. Staurosporine, a potent inhibitor of phospholipid\/Ca++ dependent protein kinase. Biochem. Biophys. Res. Commun. 135, 397\u2013402 (1986).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"30295_CR39","doi-asserted-by":"publisher","first-page":"934","DOI":"10.1038\/ncb2290","volume":"13","author":"EM Kovacs","year":"2011","unstructured":"Kovacs, E. M. et al. N-WASP regulates the epithelial junctional actin cytoskeleton through a non-canonical post-nucleation pathway. Nat. Cell Biol. 13, 934\u2013944 (2011).","journal-title":"Nat. Cell Biol."},{"key":"30295_CR40","doi-asserted-by":"publisher","first-page":"961","DOI":"10.1593\/neo.11828","volume":"13","author":"PA Knobel","year":"2011","unstructured":"Knobel, P. A., Kotov, I. N., Felley-Bosco, E., Stahel, R. A. & Marti, T. M. Inhibition of REV3 expression induces persistent DNA damage and growth arrest in cancer cells. Neoplasia 13, 961\u2013970 (2011).","journal-title":"Neoplasia"},{"key":"30295_CR41","doi-asserted-by":"publisher","first-page":"3689","DOI":"10.1529\/biophysj.104.045476","volume":"88","author":"J Guck","year":"2005","unstructured":"Guck, J. et al. Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. Biophys. J. 88, 3689\u20133698 (2005).","journal-title":"Biophys. J."},{"key":"30295_CR42","doi-asserted-by":"publisher","first-page":"312","DOI":"10.1007\/s002490050213","volume":"28","author":"M Lekka","year":"1999","unstructured":"Lekka, M. et al. Elasticity of normal and cancerous human bladder cells studied by scanning force microscopy. Eur. Biophys. J. 28, 312\u2013316 (1999).","journal-title":"Eur. Biophys. J."},{"key":"30295_CR43","doi-asserted-by":"publisher","first-page":"709","DOI":"10.1101\/pdb.top081802","volume":"2014","author":"K Bacia","year":"2014","unstructured":"Bacia, K., Haustein, E. & Schwille, P. Fluorescence correlation spectroscopy: Principles and applications. Cold Spring Harbor Protoc. 2014, 709\u2013725 (2014) (Publisher: Cold Spring Harbor Laboratory Press).","journal-title":"Cold Spring Harbor Protoc."},{"key":"30295_CR44","first-page":"138","volume":"20","author":"J Allen","year":"2011","unstructured":"Allen, J. Sample size calculation for two independent groups: A useful rule of thumb. Stat. Proc. Singap. Healthc. 20, 138\u2013140 (2011).","journal-title":"Stat. Proc. Singap. Healthc."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30295-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30295-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30295-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T13:04:33Z","timestamp":1767791073000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-30295-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,6]]},"references-count":44,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["30295"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-30295-9","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,6]]},"assertion":[{"value":"2 May 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 November 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"6 December 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"723"}}