doi:10.1016/j.microc.2024.112013engDejonghe, RinusBolea-Fernández, EduardoLores-Padin, AnaVan Acker, ThibautRua-Ibarz, AnaDe Wever, OlivierVanhaecke, FrankAn evaluation of the analytical and biological robustness of a method for quantifying iron in individual red blood cells via single-cell tandem ICP-mass spectrometryART-2024-140610This work evaluated the analytical and biological robustness of iron (Fe) determination in individual red blood cells (RBCs) via single-cell ICP-MS (SC-ICP-MS). RBCs were separated from other whole blood constituents using Ficoll-Paque™ density gradient centrifugation. While fixation with paraformaldehyde (PFA) led to RBC lysis, the use of glutaraldehyde (GA) left the RBCs intact and permitted storage of RBC suspensions in ultra-pure water for up to 16 months at 4 °C. GA-fixation also rendered the RBCs sufficiently robust to maintain integrity during their introduction into the ICP by means of nebulization of dilute suspensions. Obtaining quantitative data on a cell-per-cell basis required determination of the transport efficiency using the particle size method and external calibration against aqueous Fe standard solutions. The average Fe content per RBC obtained using SC-ICP-MS (103 fg/cell) agreed well with the value obtained using solution-based ICP-MS obtained after cell pellet digestion and with values obtained from literature. Variation of the cell number density in the suspensions analyzed between 1.5 × 105 and 6.0 × 105 cells per mL did not affect the result. Identical results from one-week interval blood drawings from healthy individuals demonstrate biological consistency. Compared to bulk analysis, the SC-ICP-MS approach offers the added value of providing information on the cell-to-cell heterogeneity in Fe content.2024http://zaguan.unizar.es/record/14575110.1016/j.microc.2024.112013http://zaguan.unizar.es/record/145751oai:zaguan.unizar.es:145751info:eu-repo/grantAgreement/EC/H2020/101034288/EU/International Fellowship Programme for Talent Attraction to the Campus of International Excellence Campus Iberus/IberusExperienceThis project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 101034288-IberusExperienceinfo:eu-repo/grantAgreement/ES/MICINN/RYC2021-031093-IMICROCHEMICAL JOURNAL 207 (2024), 112013 [8 p.]All rights reservedhttp://www.europeana.eu/rights/rr-f/info:eu-repo/semantics/embargoedAccess