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Towards single cell ICP-MS normalized quantitative experiments using certified selenized yeast.

J S F Pereira1, R Álvarez-Fernández García2, M Corte-Rodríguez2

  • 1Instituto de Química, Universidade Federal Do Rio Grande Do Sul, 91501-970, Porto Alegre, RS, Brazil; University of Oviedo, Faculty of Chemistry, Dept. of Physical and Analytical Chemistry and Instituto de Investigación Sanitaria Del Principado de Asturias (ISPA), C/Julián Clavería 8, E-33006, Oviedo, Spain.

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|August 26, 2022
PubMed
Summary
This summary is machine-generated.

A new sequential method using SELM-1 reference material improves selenium quantification in yeast cells via single-cell inductively coupled plasma-mass spectrometry (SC-ICP-MS). This approach enables accurate intracellular and extracellular selenium measurements for better experimental replication.

Keywords:
SELM-1Selenium containing cellsSelenized yeastSingle cell ICP-MS

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Area of Science:

  • Analytical Chemistry
  • Biogeochemistry
  • Cell Biology

Background:

  • Standardized protocols are crucial for reproducible single-cell inductively coupled plasma-mass spectrometry (SC-ICP-MS) experiments.
  • Selenium (Se) distribution within biological samples requires precise quantification at the single-cell level.
  • Certified Reference Materials (CRMs) are essential for method validation and inter-laboratory comparisons.

Purpose of the Study:

  • To establish a normalized procedure for replicating and comparing SC-ICP-MS experiments.
  • To quantify intracellular and extracellular selenium concentrations in yeast cells using SELM-1.
  • To evaluate the performance of sequential versus simultaneous measurement procedures for SC-ICP-MS.

Main Methods:

  • Utilized SELM-1, a selenium-enriched yeast certified reference material.
  • Employed a sequential procedure: cell washing, freeze-drying, and intracellular Se quantification via SC-ICP-MS.
  • Compared results with a direct dilution and simultaneous measurement procedure, validating cell density with flow cytometry and cell integrity with confocal microscopy.

Main Results:

  • The sequential procedure yielded accurate intracellular Se (1304 ± 48 mg kg⁻¹, 64% of certified content) and extracellular Se (412 ± 48 mg kg⁻¹, 21% of total Se), achieving 85% total Se recovery.
  • Average Se mass per yeast cell was 41.6 fg, with significant dispersion (1.6–279 fg/cell).
  • The simultaneous procedure resulted in lower total Se recovery (66%), with intracellular Se at 1024 ± 42 mg kg⁻¹ and extracellular Se at 316 ± 30 mg kg⁻¹.

Conclusions:

  • Appropriate sample preparation, particularly the sequential method, allows SC-ICP-MS to provide reliable quantitative data on intracellular and extracellular Se.
  • SELM-1 is an effective tool for normalizing data at the single-cell level, facilitating benchmarking and improvement of SC-ICP-MS studies.
  • SC-ICP-MS, when coupled with validated sample preparation, is a powerful technique for elemental analysis in biological systems.