Light element analysis of individual bacteria by x-ray microanalysis

Abstract

A method based on X-ray microanalysis (XRMA) with the transmission electron microscope for measurement of total amounts of elements in single microbial cells has been developed. All major elements in cells except hydrogen can be measured simultaneously. XRMA provided N/C ratios (means (plusmn) standard errors of the mean) for stationary-phase and growing Escherichia coli of 0.23 (plusmn) 0.01 and 0.30 (plusmn) 0.01, respectively, while CHN analysis gave values of 0.276 and 0.307, respectively, for samples from the same cultures. Analyses of free coccoliths from Emiliana huxleyi provided weight fractions close to those of CaCO(inf3): 0.35 (plusmn) 0.01, 0.15 (plusmn) 0.01, and 0.47 (plusmn) 0.01 for calcium, carbon, and oxygen, respectively. Calibration is based on monodisperse latex beads and on microdrops of defined compounds. Elements in particles in the size range from 5 fg to 500 pg are measured with a relative precision between 500 and 5,000 ppm, depending on size. As a single-cell method, XRMA avoids the shortcomings of commonly used fractionation techniques associated with bulk methods, which are based on centrifugation or filtration. On the basis of morphology and XRMA, particles may be classified more precisely into groups (e.g., biotic versus abiotic) than is possible by bulk methods. Single-cell elemental analysis may provide insight into topics like nutritional and energetic status, macromolecular composition, and (by multivariate statistics) community structure.