{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T16:00:29Z","timestamp":1791302429952,"version":"4.1.0"},"reference-count":51,"publisher":"American Chemical Society (ACS)","issue":"39","license":[{"start":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T00:00:00Z","timestamp":1790035200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100006151","name":"Basic Energy Sciences","doi-asserted-by":"publisher","award":["DE-AC02-05CH11231"],"award-info":[{"award-number":["DE-AC02-05CH11231"]}],"id":[{"id":"10.13039\/100006151","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006151","name":"Basic Energy Sciences","doi-asserted-by":"publisher","award":["MFP-10280"],"award-info":[{"award-number":["MFP-10280"]}],"id":[{"id":"10.13039\/100006151","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Formazan-based assays like MTT and XTT are widely accepted and routinely employed colorimetric methods for assessing cell viability and metabolic activity in vitro. However, there is evidence that these assays are frequently subject to interference by nano- and micro-particles through various mechanisms including adsorption, abiotic reduction, and optical interference. Formazan-based assays have previously been shown to be unsuitable for applications using carbon nanotubes, metal nano- and micro-particles, and zeolite microparticles, among others. This study examines the effect of fluorescent polystyrene (PS) beads in sizes from 200 nm to 2 \u03bcm and poly(methyl methacrylate) (PMMA) beads in sizes of 1 \u03bcm and 2 \u03bcm on the accuracy of MTT and XTT cell viability assays. Our results indicate that both PS and PMMA beads cause significant interference at sizes of 1 and 2 \u03bcm due to their attenuation spectra overlapping with the measurement wavelengths of the assays. These artifacts introduce uncertainty into viability measurements as the scattering-induced attenuation of the beads masquerades as formazan absorbance, resulting in both artificially inflated and deflated metabolic activity metrics depending on particle size and assay type. The interference of smaller 200 nm particles can be effectively corrected for by washing the plate prior to analysis. We suggest that thorough washes of the plates and additional cell-free control tests are necessary before using these assays when working with microplastics.<\/jats:p>","DOI":"10.1021\/acsomega.6c07338","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T12:01:35Z","timestamp":1790078495000},"page":"59990-60003","source":"Crossref","is-referenced-by-count":0,"title":["Interference of Microplastics with MTT and XTT Cell\nViability Assays"],"prefix":"10.1021","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-3806-8514","authenticated-orcid":true,"given":"Lydia","family":"Osborn","sequence":"first","affiliation":[{"name":"Lawrence Berkeley National Laboratory , , , ,","place":["Berkeley, California, United States, 94720"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ajay","family":"Trinh","sequence":"additional","affiliation":[{"name":"Lawrence Berkeley National Laboratory , , , ,","place":["Berkeley, California, United States, 94720"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6531-9168","authenticated-orcid":true,"given":"Behzad","family":"Rad","sequence":"additional","affiliation":[{"name":"Lawrence Berkeley National Laboratory , , , ,","place":["Berkeley, California, United States, 94720"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aeron Tynes","family":"Hammack","sequence":"additional","affiliation":[{"name":"Lawrence Berkeley National Laboratory , , , ,","place":["Berkeley, California, United States, 94720"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7643-4508","authenticated-orcid":true,"given":"Natalia","family":"Molchanova","sequence":"additional","affiliation":[{"name":"Lawrence Berkeley National Laboratory , , , ,","place":["Berkeley, California, United States, 94720"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,22]]},"reference":[{"issue":"15","key":"2026100610503825500_cit1","doi-asserted-by":"publisher","first-page":"13851","DOI":"10.1039\/D5RA09062H","article-title":"A Review on Microplastics: Sources, Environmental Fate,\nDegradation Pathways, and Analytical Identification Methods","volume":"16","author":"Kamalasekaran","year":"2026","journal-title":"RSC Adv."},{"issue":"4","key":"2026100610503825500_cit2","doi-asserted-by":"publisher","first-page":"249","DOI":"10.1021\/envhealth.3c00052","article-title":"Potential\nHealth Impact of Microplastics: A Review of Environmental Distribution,\nHuman Exposure, and Toxic Effects","volume":"1","author":"Li","year":"2023","journal-title":"Environ.\nHealth"},{"issue":"12","key":"2026100610503825500_cit3","doi-asserted-by":"publisher","first-page":"2030","DOI":"10.1002\/jat.4372","article-title":"Characterization\nof Microparticles Derived from Waste Plastics and Their Bio-Interaction\nwith Human Lung A549 Cells","volume":"42","author":"Bengalli","year":"2022","journal-title":"J. Appl. Toxicol."},{"issue":"1","key":"2026100610503825500_cit4","doi-asserted-by":"publisher","first-page":"272","DOI":"10.1039\/C9EN00523D","article-title":"Nanoplastics\nas a Potential Environmental Health Factor:\nEffects of Polystyrene Nanoparticles on Human Intestinal Epithelial\nCaco-2 Cells","volume":"7","author":"Corte\u0301s","year":"2020","journal-title":"Environ. Sci.: Nano"},{"key":"2026100610503825500_cit5","doi-asserted-by":"publisher","first-page":"04179","DOI":"10.7189\/jogh.14.04179","article-title":"Detection\nof Microplastics in Human Tissues and Organs: A Scoping Review","volume":"14","author":"Roslan","year":"2024","journal-title":"J. Glob. Health"},{"issue":"1","key":"2026100610503825500_cit6","doi-asserted-by":"publisher","first-page":"7391","DOI":"10.1038\/s41598-020-64464-9","article-title":"Potential Toxicity\nof Polystyrene Microplastic Particles","volume":"10","author":"Hwang","year":"2020","journal-title":"Sci.\nRep."},{"key":"2026100610503825500_cit7","doi-asserted-by":"publisher","first-page":"113832","DOI":"10.1016\/j.marpolbul.2022.113832","article-title":"A Review on Microplastics\nand Nanoplastics in the Environment: Their Occurrence, Exposure Routes,\nToxic Studies, and Potential Effects on Human Health","volume":"181","author":"Sangkham","year":"2022","journal-title":"Mar. Pollut. Bull."},{"issue":"1","key":"2026100610503825500_cit8","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1016\/0022-1759(83)90303-4","article-title":"Rapid Colorimetric\nAssay for Cellular Growth and Survival: Application to Proliferation\nand Cytotoxicity Assays","volume":"65","author":"Mosmann","year":"1983","journal-title":"J. Immunol. Methods"},{"key":"2026100610503825500_cit9","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1007\/978-1-61779-080-5_20","article-title":"Cell Sensitivity Assays:\nThe MTT Assay","volume":"731","author":"van Meerloo","year":"2011","journal-title":"Methods Mol. Biol."},{"issue":"3","key":"2026100610503825500_cit10","doi-asserted-by":"publisher","first-page":"911","DOI":"10.1002\/jhet.5570250340","article-title":"The Synthesis\nof XTT: A New Tetrazolium Reagent That Is Bioreducible to a Water-Soluble\nFormazan","volume":"25","author":"Paull","year":"1988","journal-title":"J. Heterocycl. 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Apitherapy Nat."},{"key":"2026100610503825500_cit14","unstructured":"An Absorbance-Based Assay for Cell Health\nand Proliferation - US.\nBioprobes 78 Journal of Cell Biology Applications. https:\/\/www.thermofisher.com\/us\/en\/home\/references\/newsletters-and-journals\/bioprobes-journal-of-cell-biology-applications\/bioprobes-78\/cyquant-xtt-cell-viability-assay-absorbance-cell-health-proliferation.html (accessed May 19, 2026)."},{"issue":"1","key":"2026100610503825500_cit15","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1016\/0022-1759(94)00277-4","article-title":"Microculture Tetrazolium\nAssays: A Comparison between Two New Tetrazolium Salts, XTT and MTS","volume":"179","author":"Goodwin","year":"1995","journal-title":"J. Immunol. 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Rev."},{"issue":"8","key":"2026100610503825500_cit28","doi-asserted-by":"publisher","first-page":"1452","DOI":"10.1016\/0959-8049(92)90542-A","article-title":"MTT Assays\nAllow Quick and Reliable Measurement of\nthe Response of Human Tumour Cells to Photodynamic Therapy","volume":"28","author":"Merlin","year":"1992","journal-title":"Eur. J. Cancer"},{"issue":"4","key":"2026100610503825500_cit29","first-page":"936","article-title":"Evaluation\nof a Tetrazolium-Based Semiautomated Colorimetric Assay: Assessment\nof Chemosensitivity Testing","volume":"47","author":"Carmichael","year":"1987","journal-title":"Cancer Res."},{"issue":"2","key":"2026100610503825500_cit30","first-page":"69","article-title":"A Critical Assessment of the Use\nof Microculture Tetrazolium Assays to Measure Cell Growth and Function","volume":"5","author":"Marshall","year":"1995","journal-title":"Growth Regul."},{"issue":"4","key":"2026100610503825500_cit31","doi-asserted-by":"publisher","first-page":"1069","DOI":"10.1021\/acs.chemrestox.0c00486","article-title":"Exposure\nof Human Lung Cells to Polystyrene Microplastics Significantly Retards\nCell Proliferation and Triggers Morphological Changes","volume":"34","author":"Goodman","year":"2021","journal-title":"Chem. Res. Toxicol."},{"key":"2026100610503825500_cit32","doi-asserted-by":"publisher","first-page":"1385387","DOI":"10.3389\/fpubh.2024.1385387","article-title":"Polystyrene\nNanoplastics Mediate Oxidative Stress,\nSenescence, and Apoptosis in a Human Alveolar Epithelial Cell Line","volume":"12","author":"Milillo","year":"2024","journal-title":"Front. Public Health"},{"key":"2026100610503825500_cit33","doi-asserted-by":"publisher","first-page":"139129","DOI":"10.1016\/j.jhazmat.2025.139129","article-title":"Small Particles, Big Problems: Polystyrene Nanoparticles\nInduce DNA Damage, Oxidative Stress, Migration, and Mitogenic Pathways\nPredominantly in Non-Malignant Lung Cells","volume":"495","author":"Ernhofer","year":"2025","journal-title":"J.\nHazard. Mater."},{"key":"2026100610503825500_cit34","doi-asserted-by":"publisher","first-page":"156089","DOI":"10.1016\/j.scitotenv.2022.156089","article-title":"Evaluation of Potential\nToxicity of Polyethylene Microplastics on Human Derived Cell Lines","volume":"838","author":"Gautam","year":"2022","journal-title":"Sci. Total Environ."},{"key":"2026100610503825500_cit35","doi-asserted-by":"publisher","first-page":"4509","DOI":"10.2147\/IJN.S381776","article-title":"Cytotoxicity\nand Genotoxicity of\nPolystyrene Micro- and Nanoplastics with Different Size and Surface\nModification in A549 Cells","volume":"17","author":"Shi","year":"2022","journal-title":"Int. J. Nanomed."},{"key":"2026100610503825500_cit36","doi-asserted-by":"publisher","first-page":"133794","DOI":"10.1016\/j.scitotenv.2019.133794","article-title":"Internalization and Toxicity: A Preliminary\nStudy of Effects of Nanoplastic Particles on Human Lung Epithelial\nCell","volume":"694","author":"Xu","year":"2019","journal-title":"Sci. Total Environ."},{"key":"2026100610503825500_cit37","doi-asserted-by":"publisher","first-page":"102738","DOI":"10.1016\/j.mtbio.2025.102738","article-title":"Polystyrene\nNanoplastics-Induced Lung Epithelial Cells Ferroptosis Promotes Pulmonary\nFibrosis via YY1\/FTL Axis","volume":"36","author":"Bu","year":"2026","journal-title":"Mater. Today Bio"},{"issue":"2","key":"2026100610503825500_cit38","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1002\/tox.23705","article-title":"Polystyrene\nMicroplastic\nParticles Induce Autophagic Cell Death in BEAS-2B Human Bronchial\nEpithelial Cells","volume":"38","author":"Jeon","year":"2023","journal-title":"Environ. Toxicol."},{"key":"2026100610503825500_cit39","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/j.jare.2023.03.003","article-title":"Polystyrene Nanoplastics\nLead to Ferroptosis in the Lungs","volume":"56","author":"Wu","year":"2024","journal-title":"J. Adv. Res."},{"issue":"11","key":"2026100610503825500_cit40","doi-asserted-by":"publisher","first-page":"1676","DOI":"10.1002\/jat.4505","article-title":"Diffractaic Acid Exhibits Thioredoxin\nReductase 1 Inhibition in Lung Cancer A549 Cells","volume":"43","author":"Gu\u0308nayd\u0131n","year":"2023","journal-title":"J. Appl. Toxicol."},{"issue":"1","key":"2026100610503825500_cit41","doi-asserted-by":"publisher","first-page":"11513","DOI":"10.1038\/s41598-019-47944-5","article-title":"Radio-Selective Effects of a Natural Occurring Muscle-Derived\nDipeptide in A549 and Normal Cell Lines","volume":"9","author":"Ybarra","year":"2019","journal-title":"Sci.\nRep."},{"issue":"7","key":"2026100610503825500_cit42","doi-asserted-by":"publisher","first-page":"8499","DOI":"10.1002\/cam4.5599","article-title":"Cigarette\nSmoke Condensate Induces Centrosome Clustering in Normal Lung Epithelial\nCells","volume":"12","author":"Thaiparambil","year":"2023","journal-title":"Cancer Med."},{"issue":"4","key":"2026100610503825500_cit43","doi-asserted-by":"publisher","first-page":"382","DOI":"10.1089\/adt.2011.391","article-title":"The XTT Cell Proliferation Assay\nApplied to Cell Layers Embedded in Three-Dimensional Matrix","volume":"10","author":"Huyck","year":"2012","journal-title":"Assay Drug Dev. Technol."},{"issue":"1","key":"2026100610503825500_cit44","doi-asserted-by":"publisher","first-page":"26","DOI":"10.2340\/0001555577026028","article-title":"Use of XTT-Assay to\nAssess the Cytotoxicity of Different Surfactants and Metal Salts in\nHuman Keratinocytes (HaCaT). A Feasible Method for in Vitro Testing\nof Skin Irritants","volume":"77","author":"Brosin","year":"1997","journal-title":"Acta Derm.-Venereol."},{"issue":"5","key":"2026100610503825500_cit45","doi-asserted-by":"publisher","first-page":"1321","DOI":"10.1021\/bm301962f","article-title":"Evaluation\nof the In Vitro Cytotoxicity of Crosslinked\nBiomaterials","volume":"14","author":"Wang","year":"2013","journal-title":"Biomacromolecules"},{"key":"2026100610503825500_cit46","volume-title":"Absorption and Scattering\nof Light by Small Particles","author":"Bohren","year":"2008"},{"key":"2026100610503825500_cit47","volume-title":"Scattering,\nAbsorption, and Emission of Light by Small Particles","author":"Mishchenko","year":"2002"},{"issue":"2","key":"2026100610503825500_cit48","doi-asserted-by":"publisher","first-page":"023110","DOI":"10.1063\/1.3068473","article-title":"Rayleigh, Mie, and\nTyndall Scatterings\nof Polystyrene Microspheres in Water: Wavelength, Size, and Angle\nDependences","volume":"105","author":"He","year":"2009","journal-title":"J. Appl. Phys."},{"key":"2026100610503825500_cit49","doi-asserted-by":"publisher","first-page":"1660","DOI":"10.1016\/j.matpr.2021.12.213","article-title":"Study of the Optical Properties of Poly (Methyl Methacrylate)\n(PMMA) by Using Spin Coating Method","volume":"60","author":"Nassier","year":"2022","journal-title":"Mater.\nToday: Proc."},{"issue":"6","key":"2026100610503825500_cit50","doi-asserted-by":"publisher","first-page":"1261","DOI":"10.1021\/nl060177c","article-title":"Oops They Did It Again! Carbon Nanotubes\nHoax Scientists in Viability Assays","volume":"6","author":"Wo\u0308rle-Knirsch","year":"2006","journal-title":"Nano Lett."},{"issue":"11","key":"2026100610503825500_cit51","doi-asserted-by":"publisher","first-page":"2163","DOI":"10.1021\/nl048715d","article-title":"Physicochemical\nProperties and Cellular Toxicity of Nanocrystal Quantum Dots Depend\non Their Surface Modification","volume":"4","author":"Hoshino","year":"2004","journal-title":"Nano Lett."}],"container-title":["ACS Omega"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsomega.6c07338","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/11\/39\/59990\/68593548\/acsomega.6c07338.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/11\/39\/59990\/68593548\/acsomega.6c07338.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T15:23:20Z","timestamp":1791300200000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acsodf\/article\/11\/39\/59990\/5435035\/Interference-of-Microplastics-with-MTT-and-XTT"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,22]]},"references-count":51,"journal-issue":{"issue":"39","published-online":{"date-parts":[[2026,9,22]]},"published-print":{"date-parts":[[2026,10,6]]}},"URL":"https:\/\/doi.org\/10.1021\/acsomega.6c07338","relation":{},"ISSN":["2470-1343"],"issn-type":[{"value":"2470-1343","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,22]]}}}