Diffuse fluids were collected from newly discovered snowblower vents at Axial Seamount in late July 2011 with the ROV Jason II using the hydrothermal fluid and particle sampler (Butterfield et al., 2004). White and orange flocculent materials were collected on the subsequent University of Washington Visions' 11 cruise, in support of the Regional Scale Nodes component of the Ocean Observatories Initiative in August 2011. White flocculent material was collected from the orifice of the Subway snowblower vent on dives R1467 (White Floc 1) and R1472 (White Floc 2) and orange flocculent material was collected on the seafloor distal to Marker 33 during dive R1472 where it coated freshly deposited basalt. All of the fluid and floc samples analyzed in this study are from a small area in the south rift zone at the southeastern edge of Axial Caldera, with the exception of background seawater which was collected outside of the caldera.
Subsamples of flocculent material were viewed under phase contrast, fixed and stained with DAPI for cell counting via epiflourescent microscopy, or fixed for scanning electron microscopy (SEM) with elemental detection system (EDS) on a Zeiss Supra 40VP. White and orange flocs were fixed with 2.5% glutaldehyde for both cell counts and SEM. Preserved whole fluids were also stained with DAPI for cell counts.
Bulk carbon and nitrogen were measured using a Thermo Scientific CN Analyzer (Model Flash 2000) from subsamples of flocculent material dried at 50–60 degrees C for 2 days. A standard curve for bulk carbon and nitrogen was made using aspartic acid as a standard and acetanilide and apple leaf as standard curve checks. Bulk sulfur was measured using a LECO S632 Sulfur Analyzer from subsamples of flocculent material dried in a dessicator for 4 days and supplemented with sterile sea sand (Fisher) to meet minimum volume requirements. A standard curve for bulk sulfur was made using coal with known sulfur content provided by LECO.
Related references:
Meyer, J.L., Akerman, N.H., Proskurowski, G. and J.A. Huber. 2013. Microbiological characterization of post-eruption "snowblower" vents at Axial Seamount, Juan de Fuca Ridge. Frontiers in Microbiology. 4:153. doi:10.3389/fmicb.2013.00153