Additional file 3 of A pipeline for targeted metagenomics of environmental bacteria

Additional file 3: Figure S3. Four pure cultures (Gramella forsetii, Maribacter forsetii, Escherichia coli, Micrococcus sp.) were fixed with 10 different fixation methods (formaldehyde 4%, 1%, 0.25%, 0.1%, ethanol, Lugol’s solution with and without thiosulfate, glyoxal with and without ethanol and unfixed). HCR-FISH was done on filtered cells and signal intensity was measured after washing the cells off the filter and analyzing them in the flow cytometer. Plotted are the green fluorescence (530/40 nm) from HCR-FISH and blue fluorescence (450/60 nm) from DAPI staining. The fluorescence intensity is given in relative units on a logarithmic scale. The background fluorescence (dotted line) was defined for 10 RU. N.A. = not analyzed due to disrupted cells.