Field-portable seawater toxicity monitoring platform using lens-free shadow imaging technology

The accidental spill of hazardous and noxious substances (HNSs) in the ocean has serious environmental and human health consequences. Assessing the ecotoxicity of seawater exposed to various HNS is challenging due to the constant development of new HNS or mixtures, and assessment methods are also limited. Microalgae viability tests are often used among the various biological indicators for ecotoxicity testing, as they are the primary producers in aquatic ecosystems. However, since the conventional cell growth rate test measures cell viability over three to four days using manual inspection under a conventional optical microscope, it is labor- and time-intensive and prone to subjective errors. In this study, we propose a rapid and automated method to evaluate seawater ecotoxicity by quantification of the morphological changes of microalgae exposed to more than 30 HNSs. This method was further validated using conventional growth rate test results. Dunaliella tertiolecta, a microalgae species without rigid cell walls, was selected as the test organism. Its morphological changes in response to HNS exposure were measured at the single cell level using a custom-developed device that uses lens-free shadow imaging technology. The ecotoxicity evaluation induced by the morphological change could be available in as little as 5 min using the proposed method and device, and it could be effective for 20 HNSs out of 30 HNSs tested. Moreover, the test results of six selected HNSs with high marine transport volume and toxicity revealed that the sensitivity of the proposed method extends to half the maximum effective concentration (EC50) and even to the lowest observed effective concentration (LOEC). Furthermore, the average correlation index between the growth inhibition test (three to four days) and the proposed morphology changes test (5 min) for the six selected HNSs was 0.84, indicating great promise in the field of various point-of-care water quality monitoring. Thus, the proposed equipment and technology may provide a viable alternative to traditional on-site toxicity testing, and the potential of rapid morphological analysis may replace traditional growth inhibition testing.