Abstract
We measured the size of Daphnia pulex (D. pulex), which is an indicator organism used for examining toxicity, under freely-swimming and stress-free conditions, made possible by the post-focusing ability of digital holography (DH). This environmental measurement is a new application of DH. This implementation, which is a key point of this research, should be applicable to real on-site use in non-sterile and unstable mechanical situations. Through trial and error, we arrived at the simplest interferometer configuration using a single beam line that can withstand vibrations, a low-coherence light source for reducing coherent noise, and single-shot image capturing. Automatic size measurement based on DH with image processing was performed. The originality of this research is the quantitative size measurement and performance estimation achieved by comparing the results with those from conventional manual eye measurement using ordinary optical microscope observation and image processing software. The estimation also allowed us to determine the proper parameters in the image processing for DH measurement.
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