A novel planar optode setup for concurrent oxygen and light field imaging: Application to a benthic phototrophic community

A novel high resolution planar optode (HiPO) for two dimensional oxygen and light-field imaging is presented. Optical cross-talk effects, limiting the precision and resolution of conventional planar optode setups, could be strongly reduced by the use of a fiber optic faceplate as sensing window. The new device, therefore, allows for accurate calculation of oxygen fluxes and respiration rates from oxygen concentration images, especially for measurements on small scales. In addition, the setup can be used to estimate the distribution of scalar irradiance within illuminated sediments or mats, thus directly linking local light availability to oxygen dynamics at high resolution. Irradiance values obtained with the HiPO were found to be in good agreement with scalar irradiance microsensor measurements in sandy sediment, and a spatial resolution of ~120 µm could be achieved. The performance of the HiPO for oxygen measurements was tested experimentally and the theoretical limit of spatial (~100 µm) and temporal (~10 s) resolution, governed by oxygen diffusion within the sensing layer, was determined by mathematical modeling. Rates of primary production and respiration in sandy sediments were calculated from the transient oxygen concentrations after perturbations in light condition showing a highly patchy distribution on submillimeter scale. These heterogeneities were clearly correlated to local irradiance within the sediment. Spatial correlations of photosynthesis and respiration were strongly dependent on incident irradiance.

[1]  David M. Ward,et al.  Photoexcretion and Fate of Glycolate in a Hot Spring Cyanobacterial Mat , 1988, Applied and environmental microbiology.

[2]  U. Franke.  Applications of planar oxygen optodes in biological aquatic systems , 2005 .

[3]  Markus Huettel,et al.  Oxygen dynamics in permeable sediments with wave‐driven pore water exchange , 2004 .

[4]  B. Jørgensen,et al.  Light penetration and light intensity in sandy marine sediments measured with irradiance and scalar irradiance fiber-optic microprobes , 1994 .

[5]  A fibre‐optic scalar irradiance microsensor: application for spectral light measurements in sediments , 1992 .

[6]  B. Jørgensen,et al.  Action spectra of microalgal photosynthesis and depth distribution of spectral scalar irradiance in a coastal marine sediment of Limfjorden, Denmark , 1993 .

[7]  M. Kühl,et al.  Functional and structural imaging of phototrophic microbial communities and symbioses , 2008 .

[8]  Ronnie N. Glud,et al.  Oxygen dynamics in the rhizosphere of Zostera marina: A two‐dimensional planar optode study , 2006 .

[9]  B. Jørgensen,et al.  Diffusive boundary layers and the oxygen uptake of sediments and detritus1 , 1985 .

[10]  Ingo Klimant,et al.  A MODULAR LUMINESCENCE LIFETIME IMAGING SYSTEM FOR MAPPING OXYGEN DISTRIBUTION IN BIOLOGICAL SAMPLES , 1998 .

[11]  Lubos Polerecky,et al.  Two-dimensional mapping of photopigment distribution and activity of Chloroflexus-like bacteria in a hypersaline microbial mat. , 2008, FEMS microbiology ecology.

[12]  H. Cypionka,et al.  Oxygen respiration by desulfovibrio species. , 2000, Annual review of microbiology.

[13]  Ronnie N. Glud,et al.  Fabrication and test of sol-gel based planar oxygen optodes for use in aquatic sediments , 2005 .

[14]  B. Jørgensen,et al.  Light-enhanced oxygen respiration in benthic phototrophic communities , 1996 .

[15]  D. D. Marais,et al.  A simple fiber-optic microprobe for high resolution light measurements: application in marine sediment , 1986 .

[16]  Ruth Shinar,et al.  Enhanced Photoluminescence of Oxygen Sensing Films through Doping with High Dielectric Constant Particles , 2007 .

[17]  L. Wales.  Functional and structural , 1999 .

[18]  Oliver Kohls,et al.  HETEROGENEITY OF OXYGEN PRODUCTION AND CONSUMPTION IN A PHOTOSYNTHETIC MICROBIAL MAT AS STUDIED BY PLANAR OPTODES , 1999 .

[19]  B. Jørgensen,et al.  Primary production of microalgae in sediments measured by oxygen microprofile, H14CO3 ‐ fixation, and oxygen exchange methods1 , 1981 .

[20]  Claudia R. Schröder,et al.  Time‐resolved pH imaging in marine sediments with a luminescent planar optode , 2006 .

[21]  N. Strömberg,et al.  Assessing an imaging ammonium sensor using time correlated pixel-by-pixel calibration , 2005 .

[22]  Hugh L. MacIntyre,et al.  Fine-scale vertical resolution of chlorophyll and photosynthetic parameters in shallow-water benthos , 1995 .

[23]  Gerhard A. Holst,et al.  Luminescence lifetime imaging with transparent oxygen optodes , 2001 .

[24]  J. Steffensen,et al.  Oxygen dynamics around buried lesser sandeels Ammodytes tobianus (Linnaeus 1785): mode of ventilation and oxygen requirements , 2007, Journal of Experimental Biology.

[25]  Mario M. Pamatmat,et al.  OXYGEN CONSUMPTION BY THE SEABED IV. SHIPBOARD AND LABORATORY EXPERIMENTS1 , 1971 .

[26]  B. Jørgensen,et al.  Photosynthesis of benthic microflora measured with high spatial resolution by the oxygen microprofile method: Capabilities and limitations of the method1 , 1983 .

[27]  J. Kromkamp,et al.  Factors controlling primary production and photosynthetic characteristics of intertidal microphytobenthos , 1998 .

[28]  Stephen J. Hawkins,et al.  Modelling patch dynamics on rocky shores using deterministic cellular automata , 2022 .

[29]  N. Revsbech,et al.  A METHOD TO IMPROVE THE SPATIAL RESOLUTION OF PHOTOSYNTHETIC RATES OBTAINED BY OXYGEN MICROSENSORS , 1998 .

[30]  V. Brotas,et al.  Responses of microphytobenthos to light: primary production and carbohydrate allocation over an emersion period , 2001 .

[31]  B. Jørgensen,et al.  The light field of microbenthic communities: Radiance distribution and microscale optics of sandy coastal sediments , 1994 .

[32]  I. Polikarpov,et al.  Diatom activity within soft sediments: behavioural and physiological processes , 2003 .

[33]  B. Hargrave.  AEROBIC DECOMPOSITION OF SEDIMENT AND DETRITUS AS A FUNCTION OF PARTICLE SURFACE AREA AND ORGANIC CONTENT , 1972 .

[34]  Ronnie N. Glud,et al.  Distribution of oxygen in surface sediments from central Sagami Bay, Japan : In situ measurements by microelectrodes and planar optodes , 2005 .

[35]  J. Serôdio,et al.  Short-term variability in the photosynthetic activity of microphytobenthos as detected by measuring rapid light curves using variable fluorescence , 2005 .

[36]  Mario M. Pam nut.  OXYGEN CONSUMPTION BY THE SEABED IV. SHIPBOARD AND LABORATORY EXPERIMENTS , 1971 .

[37]  B. Jørgensen,et al.  Microalgal photosynthesis and spectral scalar irradiance in coastal marine sediments of Limfjorden, Denmark , 1992 .

[38]  B. Biggs,et al.  PHOTOSYNTHESIS‐IRRADIANCE PATTERNS IN BENTHIC MICROALGAE: VARIATIONS AS A FUNCTION OF ASSEMBLAGE THICKNESS AND COMMUNITY STRUCTURE , 1999 .

[39]  Hao Zhang,et al.  Micro-scale biogeochemical heterogeneity in sediments : a review of available technology and observed evidence. , 2009 .

[40]  Ronnie N. Glud,et al.  Oxic microzones and radial oxygen loss from roots of Zostera marina , 2005 .

[41]  J. Koenderink.  The structure of images , 2004, Biological Cybernetics.

[42]  P. Ogilby,et al.  Oxygen Diffusion in Glassy Polymer Films: Effects of Other Gases and Changes in Pressure , 2000 .

[43]  M. Kühl,et al.  Combined Imaging of Bacteria and Oxygen in Biofilms , 2007, Applied and Environmental Microbiology.

[44]  Ingo Klimant,et al.  Planar optrodes: a new tool for fine scale measurements of two-dimensional O2 distribution in benthic communities , 1996 .

[45]  Ingo Klimant,et al.  Light Harvesting as a Simple and Versatile Way to Enhance Brightness of Luminescent Sensors , 2009, Analytical Chemistry.

[46]  E. Epping,et al.  Photosynthesis and the dynamics of oxygen consumption in a microbial mat as calculated from transient oxygen microprofiles , 1999 .

[47]  Christian Krause,et al.  Composite Luminescent Material for Dual Sensing of Oxygen and Temperature , 2006 .

[48]  N. Revsbech,et al.  Oxygen production and consumption in sediments determined at high spatial resolution by computer simulation of oxygen microelectrode data l , 1986 .

[49]  Gerhard A. Holst,et al.  Transparent oxygen optodes in environmental applications at fine scale as measured by luminescence lifetime imaging , 2002, SPIE Optics East.

[50]  Ronnie N. Glud,et al.  Platinum octaetylporphyrin based planar optodes combined with an UV-LED excitation light source: An ideal tool for high-resolution O2 imaging in O2 depleted environments , 2006 .

[51]  D. Canfield,et al.  Pathways of organic carbon oxidation in three continental margin sediments. , 1993, Marine geology.

[52]  Bernard P. Boudreau,et al.  Diagenetic Models and Their Implementation: Modelling Transport and Reactions in Aquatic Sediments , 1996 .

[53]  G. Underwood,et al.  Patterns in microphytobenthic primary productivity: Species‐specific variation in migratory rhythms and photosynthetic efficiency in mixed‐species biofilms , 2005 .

[54]  G. Barton.  The Mathematics of Diffusion 2nd edn , 1975 .

[55]  Ingo Klimant,et al.  An in situ instrument for planar O2 optode measurements at benthic interfaces , 2001 .

[56]  Markus Huettel,et al.  � 2006, by the American Society of Limnology and Oceanography, Inc. Wave tank study of particulate organic matter degradation in permeable sediments , 2022 .

[57]  J. Gattuso,et al.  Respiration in coastal benthic communities , 2004 .

[58]  Yanzhen Fan,et al.  A new ratiometric, planar fluorosensor for measuring high resolution, two-dimensional pCO2 distributions in marine sediments , 2006 .

[59]  T. Fenchel,et al.  Benthic primary production and O2-CO2 dynamics in a shallow-water sediment: Spatial and temporal heterogeneity , 2000 .

[60]  R. Glud.  Oxygen dynamics of marine sediments , 2008 .

[61]  Li-Dong Cai,et al.  Some Notes on Repeated Averaging Smoothing , 1988, Pattern Recognition.