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A variety of imaging systems are in use in oceanographic surveys, and the opto-mechanical configurations have become highly sophisticated. However, much less consideration has been given to the accurate reconstruction of imaging data. To improve reconstruction of particles captured by Focused Shadowgraph Imaging (FoSI)—a system that excels at visualizing low-optical-density objects, we developed a novel object detection algorithm to process images with a resolution of ~ 12 μm per pixel. Suggested improvements to conventional edge-detection methods are relatively simple and time-efficient, and more accurately render the sizes and shapes of small particles ranging from 24 to 500 μm. In addition, we introduce a gradient of neutral density filters as a part of the protocol serving to calibrate recorded gray levels and thus determine the absolute values of detection thresholds. Set to intermediate detection threshold levels, particle numbers were highly correlated with beam attenuation (c_p) measured independently. The utility of our method was underscored by its ability to remove imperfections (dirt, scratches and uneven illumination), and by capturing the transparent particle features such as found in gelatinous plankton, marine snow and a portion of the oceanic gel phase. 

Optical surveys of aquatic particles and their particle size spectra have become important tools in studies of light propagation in water, classification of water masses, and the dynamics of trophic interactions affecting particle aggregation and flux. Here, we demonstrate that typical settings used in image analysis vastly underestimate particle numbers due to the particle – gel continuum. Applying a wide range of threshold values to change the sensitivity of our detection system, we show that macrogels cannot be separated from more dense particles, and that a true particle number per volume cannot be ascertained; only relative numbers in relation to a defined threshold value can be reported. A quandary thus presents itself between choosing a detection threshold low enough to accurately record orders of magnitude more particles on one hand or selecting a higher threshold to yield better image quality of plankton on the other. By observing the dynamics of coagulation and dissolution steps unique to cation-bridged gels abundant in aquatic systems, we find naturally occurring gels, and microscopic particles attached to them, to cause the ill-defined particle numbers. In contrast, the slopes in particle number spectra remained largely unaffected by varying sensitivity settings of the image analysis. The inclusion of fainter particles that are not typically captured by imaging systems provides a window into the true microscale spatial heterogeneity at scales relevant to small plankton organisms and processes that are dependent on particle density such as surface-associated chemical reactions as well as particle coagulation and aggregation dynamics.