Gill filament-based EROD assay for monitoring waterborne dioxin-like pollutants in fish E. Maria J

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Gill filament-based EROD assay for monitoring
waterborne dioxin-like pollutants in fishE.
Maria Jönsson, Ingvar Brandt, and Björn
BrunströmEnviron. Sci. Technol., 36 (15), 3340
-3344, 2002. http//pubs.acs.org/cgi-bin/abstrac
t.cgi/esthag/2002/36/i15/abs/es015859a.html
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The anatomy of the gills
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Preparations

• The gill arches are dissected from the fish and
  gill filaments are cut above the septum.
• Ten gill filament tip pieces (2 mm long) are
  selected and placed in each well of a 12-well
  tissue culture plate containing Hepes-Cortland
  buffer (pH 7.7).

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Sampling and measurement

• The filament pieces are incubated in 0.7 ml of a
  buffer containing 1 mM 7-ethoxyresorufin and 10
  mM dicumarol in Hepes Cortland (pH 7.7)
• After 10 and 30 min 200 ml aliquots are sampled
  from each well.
• Fluorescence is measured at 544 nm (ex) and 590
  nm (em)
• EROD activity is calculated from the amount of
  resorufin formed between the two samplings
  divided by the time between the samplings and the
  number of filaments (10).