{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T11:12:37Z","timestamp":1788347557417,"version":"build-2803163510"},"reference-count":41,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:00:00Z","timestamp":1788307200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Dev. Biol.","JDB"],"abstract":"<jats:p>This study evaluates ChromaLIVETM, a multichromatic, mix-and-read fluorescent marker developed for continuous live-cell imaging, in the context of embryology. The aim was to evaluate whether this marker is compatible with dynamic embryonic development up to the blastocyst stage, with minimal impact on embryo viability, integrity, or normal growth of mouse embryos at the 2-cell stage that were cultured to the blastocyst stage under time-lapse imaging conditions, either in the presence or absence of ChromaLIVETM. Results showed comparable blastocyst formation rates between ChromaLiveTM and the control group, as well as similar proportions of high-quality blastocysts. Although minor but statistically significant delays were noted in two kinetics events (t3 and tSB), other developmental timings remained unaffected. Time-lapse confocal fluorescence imaging demonstrated that ChromaLIVETM dye provided stable, bright, high-resolution 3D visualizations of cellular compartments including membranes, cytoplasm and embryonic structures such as trophectoderm, inner cell mass and blastocoel throughout development. Finally, transcriptomic analyses revealed a limited set of gene-expression differences between stained and unstained embryos; although these included developmental and metabolic pathways, embryos developed to the blastocyst stage comparably to controls.<\/jats:p>","DOI":"10.3390\/jdb14030039","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T10:02:52Z","timestamp":1788343372000},"page":"39","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of a Novel Fluorescent Marker for Continuous Live Imaging of Preimplantation Embryos"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9776-7459","authenticated-orcid":false,"given":"Debbie","family":"Montjean","sequence":"first","affiliation":[{"name":"Centres d\u2019aide M\u00e9dicale \u00e0 la Procr\u00e9ation Fertilys, 1950 rue Maurice-Gauvin Street, #103, Laval, QC H7S 1Z5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Karine","family":"Audette","sequence":"additional","affiliation":[{"name":"Institut de Recherche en Immunologie et en Canc\u00e9rologie, Universit\u00e9 de Montr\u00e9al, 2950 Chemin de Polytechnique, Montr\u00e9al, QC H3T 1J4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-2491-048X","authenticated-orcid":false,"given":"Simon","family":"Mathien","sequence":"additional","affiliation":[{"name":"Institut de Recherche en Immunologie et en Canc\u00e9rologie, Universit\u00e9 de Montr\u00e9al, 2950 Chemin de Polytechnique, Montr\u00e9al, QC H3T 1J4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6094-6752","authenticated-orcid":false,"given":"Marie-H\u00e9l\u00e8ne","family":"Godin-Pag\u00e9","sequence":"additional","affiliation":[{"name":"Centres d\u2019aide M\u00e9dicale \u00e0 la Procr\u00e9ation Fertilys, 1950 rue Maurice-Gauvin Street, #103, Laval, QC H7S 1Z5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moncef","family":"Benkhalifa","sequence":"additional","affiliation":[{"name":"Centres d\u2019aide M\u00e9dicale \u00e0 la Procr\u00e9ation Fertilys, 1950 rue Maurice-Gauvin Street, #103, Laval, QC H7S 1Z5, Canada"},{"name":"M\u00e9decine et Biologie de la Reproduction et Laboratoire PERITOX, Universit\u00e9 Picardie Jules Verne, CBH-CHU Amiens Picardie, 1 Rond-Point du Professeur Christian Cabrol, 80054 Amiens, France"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"}]},{"given":"Ibrahim","family":"Bilem","sequence":"additional","affiliation":[{"name":"Saguaro Biosciences, La Centrale\u2013Espace Entrepreneurial, Universit\u00e9 Laval, Quebec, QC G1V 0A6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3509-2369","authenticated-orcid":false,"given":"Pierre","family":"Miron","sequence":"additional","affiliation":[{"name":"Centres d\u2019aide M\u00e9dicale \u00e0 la Procr\u00e9ation Fertilys, 1950 rue Maurice-Gauvin Street, #103, Laval, QC H7S 1Z5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"project-administration","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/bs.mcb.2018.03.030","article-title":"Live imaging of cell division in preimplantation mouse embryos using inverted light-sheet microscopy","volume":"145","author":"Reichmann","year":"2018","journal-title":"Methods Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1038\/nmeth.3690","article-title":"Inverted light-sheet microscope for imaging mouse pre-implantation development","volume":"13","author":"Strnad","year":"2016","journal-title":"Nat. Methods"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1016\/j.cell.2018.09.031","article-title":"In toto imaging and reconstruction of post-implantation mouse development at the single-cell level","volume":"175","author":"McDole","year":"2018","journal-title":"Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"6172","DOI":"10.1038\/s41467-020-19827-1","article-title":"In vivo biomolecular imaging of zebrafish embryos using confocal Raman spectroscopy","volume":"11","author":"Horgan","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1093\/biolre\/ioae062","article-title":"Viewing early life without labels: Optical approaches for imaging the early embryo","volume":"110","author":"Chow","year":"2024","journal-title":"Biol. Reprod."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1038\/s42003-024-06745-x","article-title":"Full-field optical coherence microscopy enables high-resolution label-free imaging of the dynamics of live mouse oocytes and early embryos","volume":"7","author":"Morawiec","year":"2024","journal-title":"Commun. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1038\/s42003-023-04934-8","article-title":"Viewing life without labels under optical microscopes","volume":"6","author":"Ghosh","year":"2023","journal-title":"Commun. Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3161","DOI":"10.1007\/s00204-021-03139-4","article-title":"Toxic effect of light on oocyte and pre-implantation embryo: A systematic review","volume":"95","author":"Khodavirdilou","year":"2021","journal-title":"Arch. Toxicol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1039\/D3CS00883E","article-title":"Bioorthogonally activated probes for precise fluorescence imaging","volume":"54","author":"Fu","year":"2025","journal-title":"Chem. Soc. Rev."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1038\/ncb2003","article-title":"A role for Rho GTPases and cell-cell adhesion in single-cell motility in vivo","volume":"12","author":"Kardash","year":"2010","journal-title":"Nat. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"DuBuc, T.Q., Dattoli, A.A., Babonis, L.S., Salinas-Saavedra, M., R\u00f6ttinger, E., Martindale, M.Q., and Postma, M. (2014). In vivo imaging of Nematostella vectensis embryogenesis and late development using fluorescent probes. BMC Cell Biol., 15.","DOI":"10.1186\/s12860-014-0044-2"},{"key":"ref_12","first-page":"102174","article-title":"Integrating ChromaLIVE\u2122 dye with an AI-powered image analysis for real-time monitoring of human mesenchymal stem cells differentiation","volume":"43","author":"Bilem","year":"2025","journal-title":"Biochem. Biophys. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"100121","DOI":"10.1016\/j.slasd.2023.10.005","article-title":"Live Cell Painting: New nontoxic dye to probe cell physiology in high content screening","volume":"29","author":"Cottet","year":"2023","journal-title":"SLAS Discov."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/S0092-8674(00)80607-8","article-title":"Endomembrane trafficking of Ras: The CAAX motif targets proteins to the ER and Golgi","volume":"98","author":"Choy","year":"1999","journal-title":"Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5931","DOI":"10.1074\/jbc.M109.066282","article-title":"Amino acid residues critical for endoplasmic reticulum export and trafficking of platelet-activating factor receptor","volume":"285","author":"Hirota","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1095\/biolreprod.116.143446","article-title":"Three-dimensional testicular organoid: A novel tool for the study of human spermatogenesis and gonadotoxicity in vitro","volume":"96","author":"Pendergraft","year":"2017","journal-title":"Biol. Reprod."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"114216","DOI":"10.1016\/j.isci.2025.114216","article-title":"High-content imaging of primary chronic lymphocytic leukemia cells predicts patient cohorts with distinct cellular drug responses","volume":"28","author":"Li","year":"2025","journal-title":"iScience"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"100268","DOI":"10.1016\/j.slasd.2025.100268","article-title":"Alternate dyes for image-based profiling assays","volume":"36","author":"Sivagurunathan","year":"2025","journal-title":"SLAS Discov."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Lippincott, M.J., Tomkinson, J., Bilem, I., Suzuki, M., Nakde, A., Endou, T., Mathien, S., Lavoie-Perusse, F., Basualto-Alarc\u00f3n, C., and Way, G.P. (2025). High-content live-cell time-lapse imaging predicts cells about to die via apoptosis. bioRxiv.","DOI":"10.1101\/2025.10.23.684203"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Liu, C., Hillsley, A., Sekhar, M., Jones, C.A., Sturm, G., Fujimori, T., Wiene, D.M., Cheng, K.W., Chandler, T., and Lin, A. (2026). A multimodal perturbation atlas defines the phenotypic resolution of cellular morphology. bioRxiv.","DOI":"10.2139\/ssrn.7361027"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Dubinsky, D., Harel, S., Bein, A., Nyska, A., Ya\u2019ari, S., Ko\u00e7, B., Anas, F., Bentwich, I., and Wolf, L. (2026). Non-invasive quantification of viability in liver spheroids using deep learning. Front. Bioeng. Biotechnol., 14.","DOI":"10.3389\/fbioe.2026.1797474"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"ESHRE Working Group on Time-Lapse Technology, Apter, S., Ebner, T., Freour, T., Guns, Y., Kovacic, B., Le Clef, N., Marques, M., Meseguer, M., and Montjean, D. (2022). Good practice recommendations for the use of time-lapse technology. Hum. Reprod. Open, 2020, hoaa008.","DOI":"10.1093\/hropen\/hoaa008"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1093\/humupd\/3.4.367","article-title":"Culture and selection of viable blastocysts: A feasible proposition for human IVF?","volume":"3","author":"Gardner","year":"1997","journal-title":"Hum. Reprod. Update"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1055\/s-2005-923389","article-title":"Quality control in human in vitro fertilization","volume":"23","author":"Gardner","year":"2005","journal-title":"Semin. Reprod. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"763","DOI":"10.1038\/11698","article-title":"Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability","volume":"17","author":"Squirrell","year":"1999","journal-title":"Nat. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"972","DOI":"10.1002\/bdr2.1984","article-title":"Toward better assessments of developmental toxicity using stem cell-based in vitro embryogenesis models","volume":"114","author":"Marikawa","year":"2022","journal-title":"Birth Defects Res."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Abdulhasan, M., Ruden, X., You, Y., Harris, S.M., Ruden, D.M., Awonuga, A.O., Alvero, A., Puscheck, E.E., and Rappolee, D.A. (2021). Using live imaging and FUCCI embryonic stem cells to rank DevTox risks: Adverse growth effects of PFOA compared with DEP are 26 times faster, 1,000 times more sensitive, and 13 times greater in magnitude. Front. Toxicol., 3.","DOI":"10.3389\/ftox.2021.709747"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"113429","DOI":"10.1016\/j.ecoenv.2022.113429","article-title":"Advanced human developmental toxicity and teratogenicity assessment using human organoid models","volume":"238","author":"Li","year":"2022","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"119604","DOI":"10.1016\/j.biomaterials.2019.119604","article-title":"DMSO impairs the transcriptional program for maternal-to-embryonic transition by altering histone acetylation","volume":"230","author":"Kang","year":"2020","journal-title":"Biomaterials"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4641","DOI":"10.1038\/s41598-019-40660-0","article-title":"DMSO induces drastic changes in human cellular processes and epigenetic landscape in vitro","volume":"9","author":"Verheijen","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_31","unstructured":"Food and Drug Administration (2021). Mouse Embryo Assay for Assisted Reproduction Technology Devices\u2014Guidance for Industry and FDA Staff, FDA."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e7","DOI":"10.1016\/j.fertnstert.2021.05.011","article-title":"Evaluating human embryo culture media performance using C57BL-6N mouse embryos","volume":"116","author":"Agrawala","year":"2021","journal-title":"Fertil. Steril."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1007\/s10815-020-01768-9","article-title":"Mouse embryo assay for human in vitro fertilization quality control: A fresh look","volume":"37","author":"Esfandiari","year":"2020","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1007\/s00441-015-2124-9","article-title":"Stress signaling in mammalian oocytes and embryos: A basis for intervention and improvement of outcomes","volume":"363","author":"Latham","year":"2016","journal-title":"Cell Tissue Res."},{"key":"ref_35","first-page":"96","article-title":"Embryo responses to stress induced by assisted reproductive technologies","volume":"86","author":"Heras","year":"2019","journal-title":"Mol. Reprod. Dev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1095\/biolreprod.110.083386","article-title":"Stress for stress tolerance? A fundamentally new approach in mammalian embryology","volume":"83","author":"Pribenszky","year":"2010","journal-title":"Biol. Reprod."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1016\/j.tem.2011.05.005","article-title":"Environmental and epigenetic effects upon preimplantation embryo metabolism and development","volume":"22","author":"Chason","year":"2011","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1038\/nbt1278","article-title":"Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence live-cell imaging","volume":"25","author":"Hoebe","year":"2007","journal-title":"Nat. Biotechnol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"jcs242834","DOI":"10.1242\/jcs.242834","article-title":"Optimizing live-cell fluorescence imaging conditions to minimize phototoxicity","volume":"133","author":"Kiepas","year":"2020","journal-title":"J. Cell Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3166","DOI":"10.1016\/j.cell.2023.06.003","article-title":"Human embryo live imaging reveals nuclear DNA shedding during blastocyst expansion and biopsy","volume":"186","author":"Skory","year":"2023","journal-title":"Cell"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"eady6402","DOI":"10.1126\/sciadv.ady6402","article-title":"Capturing aberrant cell behaviors producing defects in human embryos via live imaging","volume":"11","author":"Akizawa","year":"2025","journal-title":"Sci. Adv."}],"container-title":["Journal of Developmental Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2221-3759\/14\/3\/39\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T10:13:21Z","timestamp":1788344001000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2221-3759\/14\/3\/39"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,2]]},"references-count":41,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["jdb14030039"],"URL":"https:\/\/doi.org\/10.3390\/jdb14030039","relation":{},"ISSN":["2221-3759"],"issn-type":[{"value":"2221-3759","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,2]]}}}