Chlorophyll A Testing Part III: Flourmetry

Fluorometry:

Supplies:

  • Glass cuvettes
  • 10% HCl
  • 90% acetone
  • Kimwipes

Calibration:

  1. Check date of last compete calibration, if recent enough, proceed to check calibration with solid secondary standard; if not, skip to section V for complete calibration.
  2. Turn on Turner Trilogy Instrument (fluorometer), then open and select “start” in System Interface Software on connected computer. Once the connection is established, open “calibration” spreadsheet.
  3. Select “Ch-A” on the fluorometer screen, and select “OK” to confirm.
  4. Place the solid secondary standard into the fluorometer and close lid.
  5. Select “Measure Fluorescence Raw” and check that measured value is within 10% of the solid standard value in the “calibration” spreadsheet.
  6. If the reading is not within 10%, run a complete calibration; if within 10%, proceed to reading samples.
  7. Select “Calibrate” on the fluorometer screen, then “Use stored calibration”.
  8. Choose the most recent stored calibration, and press “select”.

Measure Samples:

  1. Choose first prepared sample, and selecting “Sample ID” in the upper-left corner of the fluorometer screen, enter an appropriate ID for the sample. Press “save”.
  2. Transfer extracted sample into cuvette; wipe cuvette with acetone using a kimwipe, place into fluorometer and close lid.
  3. Select “Measure Fluorescence” on fluorometer screen. When prompted, enter the volume of filtered water (from section II), then press “OK”.
  4. Enter the volume of solvent (90% acetone) used (from section III), then press “OK”.
  5. Ensure that cuvette is in place, then select “OK”.
  6. Once screen displays the “OK” and “Cancel” buttons again, remove cuvette from fluorometer, and add 1-2 drops of 10% HCl.
  7. Replace cuvette into fluorometer, and select “OK” on “after acidification” screen.
  8. Fluorometer will display chl a and pheo a levels, record values with correct units.
  9. Repeat steps 1-8 for each prepared sample.