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

Autor(es)

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

• Tipo de documento: Artigo Científico
• Fonte: Microchemical Journal; v. 202, p. 10-pg., 2024-05-17.

Resumo

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)

• Processo FAPESP: 19/24445-8 - EMU concedido no processo 2018/25207-0: espectrômetro de massas com plasma indutivamente acoplado
• Beneficiário: Marco Aurelio Zezzi Arruda
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
• Processo FAPESP: 18/25207-0 - Nanometalômica aplicada ao cultivo da soja para estudar os efeitos das nanopartículas relacionadas a metal
• Beneficiário: Marco Aurelio Zezzi Arruda
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 17/26131-5 - Chaperoma: estudo da relação entre a estrutura dos seus componentes e a manutenção da proteostase
• Beneficiário: Carlos Henrique Inacio Ramos
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 14/50867-3 - INCT 2014: Instituto Nacional de Ciência e Tecnologia de Bioanalítica
• Beneficiário: Marco Aurelio Zezzi Arruda
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 20/08543-7 - Um kit de ferramentas especiômicas para investigar alterações mediadas por ácidos graxos na especiação plasmática de zinco e seus efeitos fisiológicos
• Beneficiário: Marco Aurelio Zezzi Arruda
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 20/06934-9 - Caracterização e quantificação de nanopartículas de prata e sílica aplicadas durante o cultivo da soja por single particle/cell ICP-MS/MS: estudos de translocação e interação celular
• Beneficiário: Ana Beatriz Santos da Silva
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado