Developing a Cell Quenching Method to Facilitate Single Cell Mass Spectrometry Metabolomics Studies

Area of Science:

• Biochemistry
• Analytical Chemistry
• Cell Biology

Background:

• Single-cell mass spectrometry (SCMS) enables metabolite analysis in individual cells.
• Rapid metabolite turnover in cells poses challenges for sample preservation during transport and storage.
• Maintaining cellular metabolite profiles for SCMS is critical for reliable analysis.

Purpose of the Study:

• To develop and evaluate a cell preparation method for preserving cell metabolites for SCMS.
• To assess the impact of liquid nitrogen (LN2) quenching and cold storage on cellular metabolomes.
• To determine optimal conditions for metabolite preservation in SCMS workflows.

Main Methods:

• Cells were washed with a volatile salt solution.
• Rapid quenching using liquid nitrogen (LN2) was employed.
• Freeze-drying under vacuum and storage at -80 °C were utilized for preservation.
• Metabolite profiles were analyzed using SCMS.

Main Results:

• LN2 quenching effectively preserved the overall cell metabolome.
• Storage at -80 °C for 48 hours caused minor changes in quenched cell metabolites.
• Unquenched cells showed significant metabolite alterations irrespective of storage conditions.
• Pathway analysis revealed the impact of quenching omission and storage duration.

Conclusions:

• Cell quenching is essential for preserving metabolite profiles for SCMS.
• Minimizing low-temperature storage time is recommended to maintain metabolite integrity.
• The developed method facilitates extended SCMS studies with minimal metabolite alteration.