Cryogenic x-ray photoelectron spectroscopy was used to analyze the cell envelope of intact and hydrated Gram-negative bacteria of the species Pseudomonas fluorescens. We used a reference strain, DSM50090, from the German microbial culture collection, which we previously have suggested would function well as a reference strain for future XPS analyses of Gram-negative bacteria. Bacteria were grown on nutrient agar plates at room temperature, collected with a cultivation loop, and washed using phosphate buffered saline. An aliquot of the cell pellet was fast-frozen on the sample holder in the sample introduction chamber to a temperature of 103 K and kept frozen throughout the measurement. Survey spectra and high-resolution spectra of Na 1s, O 1s, N 1s, C 1s, Cl 2p, S 2p, and P 2p are reported. The spectra obtained from the analyzed cells represent a combined signal from O, N, C, and S atoms in proteins, lipids, and polysaccharides at the cell surface. Furthermore, signal from P, Na, K, and Cl atoms was present both originating from processes in the cell envelope and remnants from the wash buffer.