An evaluation of the analytical and biological robustness of a method for quantifying iron in individual red blood cells via single-cell tandem ICP-mass spectrometry

Dejonghe, Rinus ; Bolea-Fernández, Eduardo (Universidad de Zaragoza) ; Lores-Padin, Ana ; Van Acker, Thibaut ; Rua-Ibarz, Ana (Universidad de Zaragoza) ; De Wever, Olivier ; Vanhaecke, Frank

Resumen: This 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.

Idioma: Inglés

DOI: 10.1016/j.microc.2024.112013

Año: 2024

Publicado en: MICROCHEMICAL JOURNAL 207 (2024), 112013 [8 p.]
ISSN: 0026-265X

Financiación: info:eu-repo/grantAgreement/EC/H2020/101034288/EU/International Fellowship Programme for Talent Attraction to the Campus of International Excellence Campus Iberus/IberusExperience
Financiación: info:eu-repo/grantAgreement/ES/MICINN/RYC2021-031093-I

Tipo y forma: Article (PostPrint)

Área (Departamento): Área Química Analítica (Dpto. Química Analítica)