| Contributors | Affiliation | Role |
|---|---|---|
| Huettel, Markus | Florida State University (FSU) | Principal Investigator |
| Mickle, Audrey | Woods Hole Oceanographic Institution (WHOI BCO-DMO) | BCO-DMO Data Manager |
The columns used for this experiment had a length of 10 cm and a diameter of 1 cm. The sands used were carbonate sands with the grain size diameter ranges 1) 125 to 250 um and 2) 250-500 um. Prior to the experiments, the sands were cleaned through repeated washing in NaCl solution (salinity 35). For testing particle separation within these sands, seawater with a natural phytoplankton community dominated by green algae (5-10 um cell diameter) and cyanobacteria (1-5 nm cell diameter) were pumped through the sand. Computer-controlled syringe pumps pushed the seawater with the algae through the columns at a pore front velocity of 20 cm per hour. The eluent of the 2 columns were collected in a rotating sampler at 15 minute intervals. A total of 36 fractions was collected for each of the sand column for a flushing period of 9 h. The resulting samples were analyzed in a Cytoflex flow cytometer with gating adjusted to capture the green algae and cyanobacteria. Data collected include the area under the forward scatter peak, which provides information on cell size side scatter data, which are related to cell granularity. Calibration of the instrument used calibration beads with particle size ranging from 0.29 to 16.8 um. Explanations regarding flow cytometer analysis of phytoplankton samples as performed here are presented in Trask et al. (1982), best practices in Gallot et al. (2025), and details on the more recent instrumentation in Ugawa et. al (2024).
Cells penetrating through the sand columns were identified by analyzing forward (FSC) and sideward scatter (SSC) signals. The signals are used to gate and identify different cell populations. By combining information from the two scatter signals, particles can be distinguished based on their size and granularity.
- Loaded sheets 1-3 from 250402_Carb125_Cyto10cm_algae.xlsx and sheets 5-36 from the same file into the BCO-DMO system, using filename as resource name, with "" and "nd" treated as missing values
- Loaded C125-4.xlsx with data from event 4, sheet 1, named 250402_carb125_cyto10cm_algae-4 into the BCO-DMO system, same load settings as above
- Loaded sheets 1-36 from 250402_Carb250_Cyto10cm_algae.xlsx using filename as resource name into the BCO-DMO system, with "" and "nd" treated as missing values
- Concatenated 72 resources (36 from Carb125, 36 from Carb250) into a single resource; added a resource_name column to track the source sheet/file
- Renamed fields to replace hyphens and double-colon notation with underscores: FSC-H to FSC_H, FSC-A to FSC_A, SSC-H to SSC_H, SSC-A to SSC_A, FITC-H::FL1-H to FITC_H, FITC-A::FL1-A to FITC_A, PE-H::FL2-H to PE_H, PE-A::FL2-A to PE_A, APC-H::FL3-H to APC_H, APC-A::FL3-A to APC_A, APC-A750-A::FL4-A to APC_A750_A, FSC-Width to FSC_Width, APC-A750-H::FL4-H to APC_A750_H
- Split resource_name field using pattern 250402_carb(.*)_cyto10cm_algae-(.*) into two new fields: grain_size_lower_bound (lower bound of sand grain size range; 125 = 125–250 µm, 250 = 250–500 µm) and fraction_number (sequential eluent fraction number 1–36, collected at 15-minute intervals over a 9-hour flushing period); original resource_name field preserved
- Deleted resource_name field
- Reordered fields to: grain_size_lower_bound, fraction_number, Event, Time, FSC_H, FSC_A, SSC_H, SSC_A, FITC_H, FITC_A, PE_H, PE_A, APC_H, APC_A, APC_A750_H, APC_A750_A, FSC_Width
- Output written to 996646_v1_carb125_carb250_cyto10cm_algae
- Tab 4 was blank on the original spreadsheet 250402_Carb125_Cyto10cm_algae.xlsx. Submitter emailed C125-4.xlsx with the missing data. DM inserted data into the original spreadsheet before attaching it as a supplemental file.
- Upon submitter request, DM removed formatting from tab in 250402_Carb250_Cyto10cm_algae.xlsx and attached changed spreadsheet as a supplemental file.
| File |
|---|
996646_v1_carb125_carb250_cyto10cm_algae.csv (Comma Separated Values (.csv), 333.01 MB) MD5:583857691236344540c9cc4afad77be7 Primary data file for dataset ID 996646, version 1 |
| File |
|---|
250325 CytoFlex calibration .xlsx (Microsoft Excel, 22.11 MB) MD5:ffa807a20e1493e9225735cfcf30ecb6 Contains CytoFlex calibration data; CytoFlex calibration file using a set of calibration beads (0.29, 0.58, 0.79, 1.32, 2.0, 3.3, 5.2, 7.8, 10.0, 16.8 micrometer diameter)Column headers are the same as for the sample runs:Event, The sequential number of the analyzed particle (or cell)FSC-H, Forward Scatter Height; related to cell sizeFSC-A, Forward Scatter Area; a measure of total scatter intensitySSC-H, Side Scatter Height; indicates granularity or complexitySSC-A, Side Scatter Area; measures overall side scatter intensityFITC-H::FL1-H, Height of the fluorescence signal for FITC in channel 1FITC-A::FL1-A, Area of the fluorescence signal for FITC in channel 1PE-H::FL2-H, Height of the fluorescence signal for PE (Phycoerythrin) in channel 2PE-A::FL2-A, Area of the fluorescence signal for PE in channel 2APC-H::FL3-H, Height of the fluorescence signal for APC (Allophycocyanin) in channel 3APC-A::FL3-A, Area of the fluorescence signal for APC in channel 3APC-A750-H::FL4-H, Height of the fluorescence signal for APC 750 in channel 4APC-A750-A::FL4-A, Area of the fluorescence signal for APC 750 in channel 4FSC-Width, Width of the forward scatter pulse; reflects the cell’s shape or structureTime, The timestamp for the event or the elapsed time since the start of acquisition |
250402_Carb125_Cyto10cm_algae.xlsx (Microsoft Excel, 160.11 MB) MD5:2100b75dabe4b35ae3f7cd9546d07578 Includes the flow cytometer data for carbonate sand columns with the grain size diameter ranges 125 to 250 um flushed by water containing phytoplanktonColumn headers are the same as for the sample runs:Event, The sequential number of the analyzed particle (or cell)FSC-H, Forward Scatter Height; related to cell sizeFSC-A, Forward Scatter Area; a measure of total scatter intensitySSC-H, Side Scatter Height; indicates granularity or complexitySSC-A, Side Scatter Area; measures overall side scatter intensityFITC-H::FL1-H, Height of the fluorescence signal for FITC in channel 1FITC-A::FL1-A, Area of the fluorescence signal for FITC in channel 1PE-H::FL2-H, Height of the fluorescence signal for PE (Phycoerythrin) in channel 2PE-A::FL2-A, Area of the fluorescence signal for PE in channel 2APC-H::FL3-H, Height of the fluorescence signal for APC (Allophycocyanin) in channel 3APC-A::FL3-A, Area of the fluorescence signal for APC in channel 3APC-A750-H::FL4-H, Height of the fluorescence signal for APC 750 in channel 4APC-A750-A::FL4-A, Area of the fluorescence signal for APC 750 in channel 4FSC-Width, Width of the forward scatter pulse; reflects the cell’s shape or structureTime, The timestamp for the event or the elapsed time since the start of acquisition |
250402_Carb250_Cyto10cm_algae.xlsx (Microsoft Excel, 186.85 MB) MD5:484f677ebfa882ce1fc32f1dacb947af Includes the flow cytometer data for carbonate sand columns with the grain size diameter ranges 250-500 um flushed by water containing phytoplanktonColumn headers are the same as for the sample runs:Event, The sequential number of the analyzed particle (or cell)FSC-H, Forward Scatter Height; related to cell sizeFSC-A, Forward Scatter Area; a measure of total scatter intensitySSC-H, Side Scatter Height; indicates granularity or complexitySSC-A, Side Scatter Area; measures overall side scatter intensityFITC-H::FL1-H, Height of the fluorescence signal for FITC in channel 1FITC-A::FL1-A, Area of the fluorescence signal for FITC in channel 1PE-H::FL2-H, Height of the fluorescence signal for PE (Phycoerythrin) in channel 2PE-A::FL2-A, Area of the fluorescence signal for PE in channel 2APC-H::FL3-H, Height of the fluorescence signal for APC (Allophycocyanin) in channel 3APC-A::FL3-A, Area of the fluorescence signal for APC in channel 3APC-A750-H::FL4-H, Height of the fluorescence signal for APC 750 in channel 4APC-A750-A::FL4-A, Area of the fluorescence signal for APC 750 in channel 4FSC-Width, Width of the forward scatter pulse; reflects the cell’s shape or structureTime, The timestamp for the event or the elapsed time since the start of acquisition |
| Parameter | Description | Units |
| grain_size_lower_bound | Lower bound of the sand grain size range for the column from which the sample was collected; values of 125 and 250 correspond to grain size ranges of 125–250 µm and 250–500 µm | unitless |
| fraction_number | Sequential number of the eluent fraction collected from the sand column; fractions were collected at 15-minute intervals over a 9-hour flushing period, yielding 36 fractions per column (1–36). | unitless |
| Event | The sequential number of the analyzed particle (or cell) | unitless |
| Time | The timestamp for the event or the elapsed time since the start of acquisition | unitless |
| FSC_H | Forward Scatter Height; related to cell size | cell size |
| FSC_A | Forward Scatter Area; a measure of total scatter intensity | cell size |
| SSC_H | Side Scatter Height; indicates granularity or complexity | cell size |
| SSC_A | Side Scatter Area; measures overall side scatter intensity | cell size |
| FITC_H | Height of the fluorescence signal for FITC in channel 1 | cell size |
| FITC_A | Area of the fluorescence signal for FITC in channel 1 | cell size |
| PE_H | Height of the fluorescence signal for PE (Phycoerythrin) in channel 2 | cell size |
| PE_A | Area of the fluorescence signal for PE in channel 2 | cell size |
| APC_H | Height of the fluorescence signal for APC (Allophycocyanin) in channel 3 | cell size |
| APC_A | Area of the fluorescence signal for APC in channel 3 | cell size |
| APC_A750_H | Height of the fluorescence signal for APC 750 in channel 4 | cell size |
| APC_A750_A | Area of the fluorescence signal for APC 750 in channel 4 | cell size |
| FSC_Width | Width of the forward scatter pulse; reflects the cell's shape or structure | cell size |
| Dataset-specific Instrument Name | Beckman CytoFlex flow cytometer |
| Generic Instrument Name | Flow Cytometer |
| Dataset-specific Description | The resulting samples were analyzed in a Cytoflex flow cytometer with gating adjusted to capture the green algae and cyanobacteria. |
| Generic Instrument Description | Flow cytometers (FC or FCM) are automated instruments that quantitate properties of single cells, one cell at a time. They can measure cell size, cell granularity, the amounts of cell components such as total DNA, newly synthesized DNA, gene expression as the amount messenger RNA for a particular gene, amounts of specific surface receptors, amounts of intracellular proteins, or transient signalling events in living cells. Description from: http://www.bio.umass.edu/micro/immunology/facs542/facswhat.htm |
NSF Award Abstract:
This project will study the role of sandy sediments in the carbon cycle. Sandy sediments cover about one-third of the continental shelf but are not well studied. Like a sand filter, marine sands separate and trap dissolved and particulate materials as seawater moves through. These processes influence organic matter cycling in sediments. Because smaller particles travel more easily through the pore space than larger ones, they move deeper into the seabed. This causes a separation of particulate matter by size. Likewise, changes in oxygen and dissolved chemicals with depth alter the surface properties of the sediment grains. Molecules with different properties are separated based on these surface characteristics. The fates of organic carbon, dissolved nutrients and pollutants in the coastal ocean are linked to uptake by sediments, physical and microbial processes within sediments, and release. Thus, it is important to understand the processes that control the transport and accumulation of materials in the seabed. This project will study the separation of particulate and dissolved organic matter transported through marine sands. It will provide information critical for understanding the cycling of carbon and nutrients at the seafloor. Graduate and undergraduate students working on this project will receive training in marine sediment functions and state-of-the-art methods that can help solve pressing environmental issues.
The main objectives of this research are to: 1) demonstrate and quantify the chromatographic separation of organic matter in shelf sediments through the analysis of sediment cores from silicate and carbonate sand beds, 2) characterize and quantify the separation process of dissolved and particulate organic matter and identify key factors controlling this separation in the sands, and 3) quantify the influence of organic matter chromatographic separation on sedimentary oxygen consumption and dissolved inorganic carbon production in these sands. The researchers will test the hypothesis that chromatographic separation of particles and solutes takes place in both sand types but differs with respect to the substances affected and the effectiveness of the separation. Intra-grain permeability of biogenic sands can enhance separation in carbonate sands through exclusion chromatography effects. The separation process is expected to enhance decomposition activities through the concentration of degradable materials in specific sediment layers. The research objectives will be addressed with a combination of field and laboratory studies that include tracer experiments and the analysis of dissolved and particulate organic matter distribution in sand sediment cores sampled in the field. Oxygen consumption and dissolved inorganic carbon production will be measured to reveal the relevance of this process for the sedimentary degradation process. The demonstration of chromatographic separation of particles and solutes in marine sands will close a gap in our understanding of the chemical processes that govern the fate of organic matter, nutrients, and pollutants. This project will provide research training opportunities for graduate and undergraduate students. Results from this study will help improve models of the global cycles of elements that can be used for predicting global environmental change.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
| Funding Source | Award |
|---|---|
| NSF Division of Ocean Sciences (NSF OCE) |