Determination of Bacterial Cell Dry Mass by Transmission Electron Microscopy and Densitometric Image Analysis

ABSTRACT We applied transmission electron microscopy and densitometric image analysis to measure the cell volume (V) and dry weight (DW) of single bacterial cells. The system was applied to measure the DW ofEscherichia coli DSM 613 at different growth phases and of natural bacterial assemblages of two lakes, Piburger See and Gossenköllesee. We found a functional allometric relationship between DW (in femtograms) and V (in cubic micrometers) of bacteria (DW = 435 · V0.86); i.e., smaller bacteria had a higher ratio of DW to V than larger cells. The measured DW of E. coli cells ranged from 83 to 1,172 fg, and V ranged from 0.1 to 3.5 μm3(n = 678). Bacterial cells from Piburger See and Gossenköllesee (n = 465) had DWs from 3 fg (V = 0.003 μm3) to 1,177 fg (V = 3.5 μm3). Between 40 and 50% of the cells had a DW of less than 20 fg. By assuming that carbon comprises 50% of the DW, the ratio of carbon content to Vof individual cells varied from 466 fg of C μm−3 forVs of 0.001 to 0.01 μm3 to 397 fg of C μm−3 (0.01 to 0.1 μm3) and 288 fg of C μm−3 (0.1 to 1 μm3). Exponentially growing and stationary cells of E. coli DSM 613 showed conversion factors of 254 fg of C μm−3 (0.1 to 1 μm3) and 211 fg of C μm−3 (1 to 4 μm3), respectively. Our data suggest that bacterial biomass in aquatic environments is higher and more variable than previously assumed from volume-based measurements.