Dataset: primary_production
Deployment: COOK19MV

 dates:           24 January 2002 to 20 February 2002  (20020124-20020220) 
 location:        N: -52.385  S: -66.612  W: -172.693  E: -166.939 
 project/cruise:  SOFeX/MV

12 February 2008: Prepared for OCB data system by Dave DuBois (WHOI) Cyndy Chandler, OCB DMO (WHOI) from documentation contributed by originating PI, data analysts and technicians.

Original Excel file downloaded from MBARI: copy of original Excel file

Contact: Anna Hilting (Duke University Marine Laboratory)

R/V Melville Extracted Chlorophyll Methodology

Please direct questions to Sara Tanner (tanner@mlml.calstate.edu) or Jodi Brewster (jbrewster@mlml.calstate.edu)

Water samples were collected from 12 depths on the CTD Rosette and 8 depths on the TM Rosette. The TM Rosette depths were chosen at the 100, 45, 30, 16, 10, 5, 1, and 0.1 percent light levels (so phytoplankton production can be related to phytoplankton biomass) (Evans et al 1987). The CTD also had 2 more depths scattered between .1 and 100 percent and one each at 200m and 300m. The water from the CTD and TM rosettes was collected using opaque brown bottles in 250, 500, 1000, and 2000 ml or white 100 ml bottles. The differing volumes depended upon the depth of the sample and whether the samples were taken within the patch or not. Sampling from the TM Rosette was done with gloves. Each bottle was rinsed three times with the sample water before filling to the neck of the bottle.

A Whatman G/FF glass Fiber Filter, (~0.7um) Polycarbonate 5 um filter, or Polycarbonate 20 micron filter was placed in a 25 mm diameter Gelman filter holder. Water was pumped through the filter, being careful the vacuum pressure did not get above 6 psi to avoid cell lyse. After filtration, the vacuum was turned off and the filter was added with forceps to a tube filled with 8 ml of 90% acetone. The tube was labeled and stored in a freezer for a minimum of 24 hours.

After the minimum 24 hours extraction time, the filter was removed from the tube and the tube was wiped down with Chem Wipes. The fluorescence of the chlorophyll extracts were read on a 10AU Turner Designs fluorometer. Two drops of 10 % HCl was added and the fluorescence was reread and recorded again. The "before" and "after" readings were plugged into equation chl-a = K * (Rb-Ra) * (vol ext/vol filtered)*dil to calculate chlorophyll a values.

A standard made from Sigma Chl-a in 90% acetone was calibrated on a spectrophotometer and used to calibrate the fluorometer at the beginning, mid and end of the cruise. Due to the fact the fluorometer drifted both ± according to the solid standard, and a high correlation was found between the low solid standard and the calibration curve, Chlorophyll-a values were corrected using the ratio of the low solid standard.

 dates:           24 January 2002 to 20 February 2002  (20020124-20020220) 
 location:        N: -52.385  S: -66.612  W: -172.693  E: -166.939 
 project/cruise:  SOFeX/MV

12 February 2008: Prepared for OCB data system by Dave DuBois (WHOI) Cyndy Chandler, OCB DMO (WHOI) from documentation contributed by originating PI, data analysts and technicians.

Original Excel file downloaded from MBARI:
copy of original Excel file

Contact: Anna Hilting (Duke University Marine Laboratory)

R/V Melville Extracted Chlorophyll Methodology



Please direct questions to Sara Tanner (tanner@mlml.calstate.edu) or Jodi Brewster (jbrewster@mlml.calstate.edu)





Water samples were collected from 12 depths on the CTD Rosette and 8
depths on the TM Rosette. The TM Rosette depths were chosen at the 100,
45, 30, 16, 10, 5, 1, and 0.1 percent light levels (so phytoplankton
production can be related to phytoplankton biomass) (Evans et al 1987).
The CTD also had 2 more depths scattered between .1 and 100 percent and
one each at 200m and 300m. The water from the CTD and TM rosettes was
collected using opaque brown bottles in 250, 500, 1000, and 2000 ml or
white 100 ml bottles. The differing volumes depended upon the depth of
the sample and whether the samples were taken within the patch or not.
Sampling from the TM Rosette was done with gloves. Each bottle was
rinsed three times with the sample water before filling to the neck of
the bottle.



A Whatman G/FF glass Fiber Filter, (~0.7um) Polycarbonate 5 um filter, or
Polycarbonate 20 micron filter was placed in a 25 mm diameter Gelman filter
holder. Water was pumped through the filter, being careful the vacuum
pressure did not get above 6 psi to avoid cell lyse. After filtration, the
vacuum was turned off and the filter was added with forceps to a tube
filled with 8 ml of 90% acetone. The tube was labeled and stored in a
freezer for a minimum of 24 hours.

After the minimum 24 hours extraction time, the filter was removed from
the tube and the tube was wiped down with Chem Wipes. The fluorescence of
the chlorophyll extracts were read on a 10AU Turner Designs fluorometer.
Two drops of 10 % HCl was added and the fluorescence was reread and
recorded again. The "before" and "after" readings were plugged into
equation chl-a = K * (Rb-Ra) * (vol ext/vol filtered)*dil to calculate
chlorophyll a values.



A standard made from Sigma Chl-a in 90% acetone was calibrated on a
spectrophotometer and used to calibrate the fluorometer at the beginning,
mid and end of the cruise. Due to the fact the fluorometer drifted both ±
according to the solid standard, and a high correlation was found between
the low solid standard and the calibration curve, Chlorophyll-a values were
corrected using the ratio of the low solid standard.