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IN SITU OPTICAL METHODS for CHLOROPHYLL ESTIMATION in the SEA a Thesis Presented to the Faculty of the Department of Biology

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IN SITU OPTICAL METHODS for CHLOROPHYLL ESTIMATION in the SEA a Thesis Presented to the Faculty of the Department of Biology ---IN SITU OPTICAL METHODS FOR CHLOROPHYLL ESTIMATION IN THE SEA A Thesis Presented to The Faculty of the Department of Biology San Jose State University In Partial Fulfillment of the Requirements for the Degree Master of Arts by Jeffrey W. Nolten December 1980 Acknowledgements This research was supported by National Oceanographic and Atmospheric Administration Grant No. 04-m~Ol-129. I 1-.rould like to thank Dennis Clark of NOAA and Robert Wrigley of NASA for free access to their cruise data and for their time and advise. A thank you to Lynn Krasnow of MLML for the art work. I would like to thank the students and staff of Moss Landing Marine Laboratories for making my tenure there a period of personal growth and fulfillment. I thank my wife Donna for her patience and understanding. A special thanks goes to Dr. William Broenkow of MLML, for without his guidance this work would never have been possible. ii CONTENTS Figures and Tables •.....•.....•...•.••.....••.•.•.•...•••.•.•... iv Introduction ........•...•...........•.......••..•.•......•..•.•. 1 Data Sources and ~1ethods ..........................•...•..•..•... 8 Results and Discussion .......•••...........•...•.......•••...•.. 11 The Secchi Disk •...................•.............•............ 15 Munsell Color .........•.....••....•.•.............•...•...•.•. 22 Conclusion •............•....•.•.............•..........••....... 38 References ........................•..•..............••......•... 41 iii Figures Figure 1. Attenuation of filtered seawater and absorption of natural phytoplankton as a function of wavelength ......... 3 Figure 2. Inverse Secchi depth vs. chlorophyll concentration .......... 16 Figure 3. Diffuse attenuation coefficient vs. chlorophyll con~entrati on ............................................. 19 Figure 4. Inverse of Secchi depth vs. chlorophyll concentration .............•............................... 20 Figure 5. Inverse of Secchi depth vs. diffuse attenuation coefficient at 440nm ...................................... 23 Figure 6. Inverse of Secchi depth vs. diffuse attenuation coefficient at 520nm ...................................... 24 Figure 7. CIE color matching functions ................................ 26 Figure 8. Chromaticity coordinates for ocean colors ................... 29 Figure 9. Chromaticity coordinates for ocean colors and Munsell hues. .. .. .. .. .. .. 33 Figure 10. Comparison of chlorophyll predictions by several optical methods ........................................... 39 Tables Table 1. Paired replicate chlorophyll samples determined photometrically and fluorometrically ........................ 12 Table 2. Linear regressions for various optical parameters ............. 13 Table 3. Regressions of optical parameters vs. ocean color ............. 31 Table 4. Proposed selection of Munsell colors .......................... 36 iv INTRODUCTION The photosynthetic pigments, chlorophyll ~'~'£and associated accessory pigments, are the only significant means of absorbing the sun•s light energy to fuel life processes. It follows that all methods of chlorophyll determination are based on the modification of light ·passing through the medium containing these pigments. Measures of the chlorophyll content of marine waters have long been used as an indi­ cator of phytoplankton standing stock and more generally, of the productivity of marine food chains as a whole. Traditional methods for measurement have included using the absorbance or fluorescence of chlorophyll pigments extracted from discrete marine water samples (Richards with Thompson 1952, Yentsch and Menzel 1963). Lorenzen (1966) introduced an extension of the fluorometric method which per­ mitted continuous horizontal or vertical chlorophyll profiles. When a beam of light passes through a medium, its radiance is attenuated where L0 is the inherent radiance and Lr is the apparent radiance after passing through path length, r (symbols follow Jerlov 1976); cis the beam attenuation coefficient which may be divided into attenuation due to absorption, a, and to scattering, b. These two components are additive, such that c = a + b. [1] Both absorption and, to a lesser extent, scattering are wavelength dependent. For water, the absorption coefficient is very strong for 2 all wavelengths except in the range of visible light from about 350 to 750 nm. This absorption minimum is so striking in the visible wave- lengths that while ultraviolet and infrared light cannot penetrate more than a meter or two in the clearest waters, blue light has been detected photometrically at depths of 600 m (Duntley 1963). Scattering -4 by water mol~cules is wavelength dependent on the order of A and com- pared with absorption, only significant in the visible wavelengths. Sixty percent of the total attenuation for blue light in pure sea water is due to scattering (Duntley 1963). It is precisely in this visible light window for water that chlorophyll absorbs most strongly (Figure 1). The absorption spectra for the pigments in various species of phytoplankton appear quite similar (Yentsch 1960) with a broad maximum at about 440 nm, a local minima between 550 nm and a smaller peak at about 675 nm. The peak in the 650 nm range is relatively sharp and varies slightly in location for chlorophylls ~' ~and£· It is thus the basis for the spectro­ photometric method of chlorophyll determination (Richards with Thompson, 1952). In sea water, this smaller peak is in the range of increasing absorption by water molecules. While dissolved chlorophyll, as ex- tracted in acetone, does not significantly scatter, a component of scattering is associated with the phytoplankton cells containing chlorophyll in natural waters. Consider now the addition of suspended particles such as phyto­ plankton, zooplankton, organic detrital material or inorganic sedi- ments. The absorption and scattering components of these particles must be added to those for sea water. Additional1y, 11 yellow sub- 3 0.24 1.0 -- - filtered seawater phytoplankton 0.20 I I 0.8 ·.E- I -E 0.16 I ' 0.6 ' -z I -z 0 0 1-- 0.12 I i= <:( I a.. ~ 0:: z I 0.4 0 w (/) 1- 0.08 I m / <t ~ I I 0.2 0.04 / "' ""'- 0.00 0.0 400 450 500 550 600 650 700 WAVELENGTH Cn m) Figure 1. Attenuation of filtered seawater and &bsorption of natural phytoplankton as a function of wavelength '(after Yentsch, 1960). 4 stance" (dissolved organic matter) (Jerlov 1976) adds its component of absorbance. Thus, equation [1] becomes where subscripts, w~ indicate a coefficient for water, p for particles andy for 11 yellow substance". Separating out the absorption due to ·chlorophyll 'from the total attenuation as measured by a beam trans- missometer could prove difficult if interference from other scattering particles or "yellow substance" is significant. Thus far, we have considered factors affecting the attenuation of a collimated beam. Sun light in the sea is diffuse and the assumption of collimation no longer holds. The irradiance due to scattered light reaching a point at .some depth can be the major contributor to the total irradiance at that point. An irradiance meter can be used to determine the diffuse attenuation of downwelled irradiance, KE. Smith and Baker (1977) have divided this into the components of atten­ uation due to water, chlorophyll and other factors: Ktotal -K+K+K- ·w c other· [2] However, as irradiance attenuation is an apparent rather than an inherent optical property of ocean water (Preisendorfer, 1961), this decomposition is not strictly permissible. H¢jerslev (1980) suggests that light attenuation in the upper few meters of the water column is due primarily to absorption and thus equation [2] is approximately at = a + a + a . w c 0 Absorption is an inherent property and therefore appropriate for s in this manner. 5 Yentsch (1960, 1971) has proposed a method for deriving informa­ tion related to the absorption by chlorophyll from radiance upwelled from the sea surface. Light returning to the surface is mostly scat­ tered sun light, emission from fluorescence or bioluminescence being relatively insignificant (Yentsch 1971; but see Gordon 1979). Further, . Yentsch suggests that, since scattering is rather wavelength indepen­ dent when compared to absorption, the ratios of upwelled nadir radiances or irradiances at different wavelengths will be primarily affected by absorption. Considering the relatively uniform low absorbance by water in the 440 to 550 nm range as compared with the change in chloro­ phyll absorption in the same range (Figure 1), the ratio of upwelled radiances L440;L550 will strongly reflect changes in the relative contribution of chlorophyll to the total absorbance of the water column. Absorbance at 440 nm by 11 yellow substance 11 will be an inter­ fering factor where present. This method offers the most promise for remote sensing, but Yentsch (1960) stresses the need for narrow band width for selecting the 440 and 550 nm wavelengths by radiometers. Recently, with the ability to mount precision radiometers aboard aircraft or satellite platforms, increasing attention has been paid to deriving chlorophyll content from upwelled optical information (visible radiance from the sea surface). While the accuracy of prediction obtained thus far has been limited to a quarter of a log of the chloro­ phyll content (Morel and Gordon 1979), the speed of measurement and the area of coverage possible with these sensors yield valuable information (Clarke, et ~-· 1970; Arvesen, et ~., 1973; Mueller, 1976). In a single two-minute pass, the Coastal Zone Color Scanner 6
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