Ms. Lingyan Kan Profile

Lingyan Kan

at Qingdao Institute of Bioenergy and Bioprocess Technology CAS

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Proceedings Article | 30 April 2024 Paper

Underwater in-situ plankton imaging system based on microscopic optical tweezers

Yanfu Zhu, Lingyan Kan, Jia Yu, Huiping Liu, Yuandong Li, Caiyun Miao

Proceedings Volume 13154, 131540D (2024) https://doi.org/10.1117/12.3017799

KEYWORDS: Optical tweezers, Imaging systems, Detection and tracking algorithms, Particles, Holography, Ocean optics, Objectives, Modulation, Holograms, Microscopes

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Marine plankton play an irreplaceable role in the marine ecological environment, and it is also an important and popular research field in biology and oceanography. Microscopic imaging system, with the outstanding advantages of high resolution and high magnification, is one of the most direct and effective in situ observation and research tools for marine plankton. To carry out the detection and research of marine plankton more efficiently and reduce the impact of its free movement on in-situ observation, this paper proposes to combine the optical tweezers with the microscopic imaging technology to design a microscopic optical tweezers based underwater in-situ observation system for plankton. The system uses holographic optical tweezers to enable the capturing of plankton in the water body, which allows it positioned stably and imaged clearly within the field of view.

The work described in this paper built a holographic microscopic optical tweezers system in the laboratory and wrote an algorithm for generating phase maps based on diffraction-computed holography, thus break through the limitation that the traditional optical tweezers system can only capture the target at the center of the optical path and realizing free manipulation of targeting plankton in the field of view. Meanwhile, this paper also miniaturizes the laboratory system and completes the design including the underwater chamber and related accessories so that the system can work well underwater. The experiments show that the system can stabilize and image plankton samples well in both static water and flow field, which can meet the requirements of underwater in-situ experiments, indicating that the method has great practical value and application prospects in solving the problem of high-quality in-situ microscopic observation.

Proceedings Article | 9 October 2021 Poster + Paper

Single-cell sorting of marine plankton based on micro-optical tweezers

Caiyun Miao, Jia Yu, Lingyan Kan, Huiping Liu, Yuandong Li

Proceedings Volume 11900, 119002J (2021) https://doi.org/10.1117/12.2601557

KEYWORDS: Optical tweezers, Particles, Ocean optics, Oceanography, Luminescence, Objectives, Microfluidics, Imaging systems, Microscopes, Water

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Marine plankton are micro and diverse, and most of them are still non-cultivable. So single-cell analysis, focused on the study of diversity and heterogeneity of their cells, has become a hot issue in marine microbiology research, which relies on sound single-cell preparation technology for upstream operations. By combining micro-optical tweezers technology with microfluidic droplet wrapping technology, we developed a fluorescence microscopic optical tweezers system for plankton sorting. Our system consists of a micro-optical tweezers module, a fluorescence module, an imaging module and a sorting module. Using this system, we conduct single-cell observation, monitoring, capture manipulation, separation and extraction of microplankton in seawater samples. The specific process is as follows: 1. Prepare samples, including grading filtration and enrichment of seawater sampled in Qingdao offshore. 2. Observe and select the target single-cell with a specific sorting chip on the microscopic optical tweezers single-cell sorting system. And perform visualized microdroplet single-cell sorting through optical tweezers capture manipulation. 3. Take out the target living single-cell after forming microdroplets. Using our experimental system, we are able to provide precise technical support for downstream experiments including single-cell culture, sequencing and monoclonality. In addition, our method has the advantages of flexible operation, intact cells and visualization as "what you see is what you get". Meanwhile, it has a 100% single-cell acquisition rate while maintaining the high viability of the sorted cell.

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