A sample preparation for quantitative determination of magnesium in individual lymphocytes by electron probe X‐ray microanalysis

Abstract

We present a sample preparation method for measuring magnesium in individual whole lymphocytes by electron probe X‐ray microanalysis. We use Burkitt's lymphoma cells in culture as the test sample and compare X‐ray microanalysis of individual cells with atomic absorption analysis of pooled cell populations. We determine the magnesium peak‐to‐local continuum X‐ray intensity ratio by electron probe X‐ray microanalysis and calculate a mean cell magnesium concentration of 39± 19 mmol/kg dry weight from analysis of 100 cells. We determine a mean cell magnesium concentration of 34 ±4 mmol/kg dry weight by atomic absorption analysis of pooled cells in three cell cultures. The mean cell magnesium concentrations determined by the two methods are not significantly different. We find a 10% coefficient of variation for both methods of analysis and a 30% coefficient of variation in magnesium concentration among individual cells by electron probe X‐ray microanalysis. We wash cells in ammonium nitrate for microanalysis or in buffered saline glucose for atomic absorption analysis. We find cells washed in either solution have the same cell viability (85%), recovery (75%), cell volume (555 μm³) and cytology. We air dry cells on thin film supports and show by magnesium X‐ray mapping that magnesium is within the cells. We conclude that: (a) our microanalysis cell preparation method preserves whole intact lymphocytes; (b) there is no systematic difference in results from the two methods of analysis; (c) electron probe X‐ray microanalysis can determine the variation in magnesium concentration among individual cells.