Resazurin-based aerobic respiration assay

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Method synopsis

Measurement of oxygen consumption rates in pore water samples using resazurin as an indicator dye. Samples preserved at 4°C for 24 hours before spectrofluorometric analysis to determine aerobic respiration rates and calculate Damköhler numbers.

Synthesized from method descriptions across 2 papers using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Hyporheic Zone Microbiome Assembly Is Linked to Dynamic Water Mixing Patterns in Snowmelt-Dominated Headwater Catchments (2019), Journal of Geophysical Research: Biogeosciences. Implementations in other papers (listed below) may differ.

1. Conduct aerobic respiration assays
   Unfiltered pore water samples from 10-, 20-, and 50-cm depths from each sampling location and season were preserved (refrigerated at 4°C for 24 hr) for a resazurin-based aerobic respiration assay to track oxygen consumption, which was analyzed using a Horiba Fluoromax 3 spectrofluorometer. Aerobic respiration measurements were then used to calculate Damköhler numbers to determine the dominance of the oxygen respiration reaction relative to transport processes.
   Quantities: Samples from 3 depths (10, 20, 50 cm) at each of 3 locations for each of 3 seasonsDuration: Samples refrigerated at 4°C for 24 hours before analysisConditions: Base flow characterization performed in October 2016 instead of August 2016 due to infrastructure constraints
   Equipment: Horiba Fluoromax 3 spectrofluorometer, resazurin

Typical Equipment

• Horiba Fluoromax 3 spectrofluorometer
• resazurin

Output Measurements

• oxygen consumption rates
• aerobic respiration rates
• Damköhler numbers

Papers Using This Protocol (2)

Species Studied With This Protocol

Places Where Used