Optimizing the performance of single-cell ICP-MS/MS for Fe and Zn determination in human umbilical vascular endothelial cells

Author(s)

Leal, Ketolly Natanne da Silva ; da Silva, Ana Beatriz Santos ; Aragao, Annelize Zambon Barbosa ; Ramos, Carlos Henrique Inacio ; Stewart, Alan J. ; Arruda, Marco Aurelio Zezzi

• Document type: Journal article
• Source: Microchemical Journal; v. 202, p. 10-pg., 2024-05-17.

Abstract

Inductively coupled plasma-mass spectrometry operated in a single-cell mode (scICP-MS/MS) has emerged as a technique that allows measurement of metal concentration at the cellular level. The approach is made possible by the high sensitivity of modern instrumentation. However, specific applications may be limited by spectral and non-spectral interference that can occur due to the complexity of the matrix that requires to be analysed. As such, the success of this approach requires adequate sample preparation, allowing the stability of the cells to be preserved before introducing them into the plasma. In this context, a method was developed to measure Zn and Fe concentrations in individual human umbilical vein endothelial cells (HUVEC) using scICP-MS/MS. As a strategy, the suspension of HUVECs was prefixed with paraformaldehyde aiming to achieve experimental stability and repeatability without significantly affecting the background signal. Moreover, different dwell times (3, 5 and 10 ms) were assessed to optimize the instrumental parameters. As a result, the best performance for Zn and Fe measurement by scICP-MS/MS in terms of sensitivity and accuracy was obtained with the dwell time of 3 ms. To the best of our knowledge, the concentrations of Fe and Zn in fixed single endothelial cells has not been explored previously using scICP-MS. Here, the developed scICP-MS/MS method was successfully used to examine the influence of different culture conditions on Fe and Zn uptake in fixed single HUVEC cells. (AU)

• FAPESP's process: 19/24445-8 - EMU granted in process 2018/25207-0: inductively coupled plasma mass spectrometer
• Grantee: Marco Aurelio Zezzi Arruda
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 18/25207-0 - Nanometallomics applied to soybean cultivation for studying the effects of metal-related nanoparticles
• Grantee: Marco Aurelio Zezzi Arruda
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 17/26131-5 - The chaperome: study of the relationship of the structure of its components and the maintenance of proteostasis
• Grantee: Carlos Henrique Inacio Ramos
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 14/50867-3 - INCT 2014: National Institute of Science and Technology in Bioanalysis
• Grantee: Marco Aurelio Zezzi Arruda
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 20/08543-7 - A speciomic toolkit to investigate fatty acid-mediated changes in plasma zinc speciation and their physiological effects
• Grantee: Marco Aurelio Zezzi Arruda
• Support Opportunities: Regular Research Grants
• FAPESP's process: 20/06934-9 - Characterization and quantification of silver and silica nanoparticles applied in soybean cultivation by a single part/cell ICP-MS/MS: translocation and cell interaction studies
• Grantee: Ana Beatriz Santos da Silva
• Support Opportunities: Scholarships in Brazil - Post-Doctoral