LN2 snap freezing has been widely used in biological research . Instead of using cold organic solvents containing buffer salts, quenching by LN2 seems more suitable for SCMS studies because LN2 can immediately stop metabolomic activities without leaving residual nonvolatile salts after LN2 evaporation. However, previous studies showed LN2 snap freezing often led to cell membrane damage , which is undesirable for SCMS studies. To prevent cell membrane damage in LN2 quenching, a method combining fast filtration, NaCl solution washing, and LN2 freezing was employed for the metabolome analysis of suspended animal cells . Briefly, cell suspension was quickly filtered by a filter (glass fiber filter disk) using a vacuum, and the filter containing cells was rinsed by cold iso-osmotic NaCl solution to remove residual culture medium and then frozen in LN2. This method is effective to retain metabolites, including those with high turnover rates, and mitigate cell membrane damage, but it is unlikely suitable for SCMS studies because of challenges to isolate cells for experiment and matrix effect due to remaining nonvolatile salts. In addition to LN2 quenching, sample storage in a -80 °C freezer is commonly used to preserve cells and tissues prior to analysis. However, the influence of storage at -80 °C on metabolite profiles of single cells has not been previously reported. There is a crucial need for developing new cell quenching methods for robust SCMS metabolomics studies.