Holographic microscopy has emerged as a tool for in situ imaging of microscopic organisms and other particles in the marine environment: appealing because of the relatively larger sampling volume and simpler optical configuration compared to other imaging systems. However, its quantitative capabilities have so far remained uncertain, in part because hologram reconstruction and image recognition have required manual operation. Here, we assess the quantitative skill of our automated hologram processing pipeline (CCV Pipeline), to evaluate the size and concentration measurements of environmental and cultured assemblages of marine plankton particles, and microspheres. Over 1 million particles, ranging from 10 to 200 μm in equivalent spherical diameter, imaged by the 4‐Deep HoloSea digital inline holographic microscope (DIHM) are analyzed. These measurements were collected in parallel with a FlowCam (FC), Imaging FlowCytobot (IFCB), and manual microscope identification. Once corrections for particle location and nonuniform illumination were developed and applied, the DIHM showed an underestimate in ESD of about 3% to 10%, but successfully reproduced the size spectral slope from environmental samples, and the size distribution of cultures (Dunaliella tertiolecta, Heterosigma akashiwo, and Prorocentrum micans) and microspheres. DIHM concentrations (order 1 to 1000 particles ml−1) showed a linear agreement (r2 = 0.73) with the other instruments, but individual comparisons at times had large uncertainty. Overall, we found the DIHM and the CCV Pipeline required extensive manual correction, but once corrected, provided concentration and size estimates comparable to the other imaging systems assessed in this study. Holographic cameras are mechanically simple, autonomous, can operate at very high pressures, and provide a larger sampling volume than comparable lens‐based tools. Thus, we anticipate that these characterization efforts will be rewarded with novel discovery in new oceanic environments.
With the ongoing, exponential increase in ocean data from autonomous platforms, satellites, models, and in particular, the growing field of quantitative imaging, there arises a need for scalable and cost-efficient visualization tools to interpret these large volumes of data. With the recent proliferation of consumer grade head-mounted displays, the emerging field of virtual reality (VR) has demonstrated its benefit in numerous disciplines, ranging from medicine to archeology. However, these benefits have not received as much attention in the ocean sciences. Here, we summarize some of the ways that virtual reality has been applied to this field. We highlight a few examples in which we (the authors) demonstrate the utility of VR as a tool for ocean scientists. For oceanic datasets that are well-suited for three-dimensional visualization, virtual reality has the potential to enhance the practice of ocean science.
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