Development of implantable fluorescent biosensors

Development of implantable fluorescent biosensors

Reducing the anthropogenic load on Lake Baikal is an important task for Lake Baikal Foundation. Due to industrial activity on the Baikal natural territory, the content of heavy metals in the reservoir has increased significantly, which leads to the immediate death of representatives of the lake’s fauna. To combat the environmental problems of the reservoir, it is necessary to identify the level of water pollution, analyze and monitor the content of heavy metals in the lake, as well as track the impact of harmful chemical elements on the well-being and life of microorganisms in lake Baikal. Thanks to the project to develop implantable fluorescent biosensors, Lake Baikal Foundation is one step closer to its goal.

Under the leadership of Anton Gurkov, the winner of the Baikal initiative competition, new ways of monitoring the ecological state of lake Baikal and its inhabitants were studied and implemented. As part of the study, a new tool was developed that allowed you to quickly track the content of heavy metals directly in the tissues of a living organism. The implantable optical microsensor made it possible to see the reaction of amphipods to the pollution of lake Baikal, which in the future will allow us to adjust the existing water quality standards in reservoirs.

Goal:

  • to develop a new type of implantable optical biosensors based on live yeast cells that are used to track the content of copper ions directly in the circulatory system of Baikal endemic amphipods.

Targets:

  • to provide grant support to Irkutsk state University (IGU) for the implementation of the corresponding project;
  • as part of the grant support, to carry out the following works in February-December 2019:

1) to select coatings that simultaneously reduce the immune response of crustaceans to microsensors and exclude the spread of yeast into the body of amphipods;

2) to obtain a genetically modified yeast strain that allows determining the concentration of copper in the medium by their fluorescence spectrum;

3) to work out the methods of using yeast in the hemocele of amphipods.

What was done:

  • The Foundation’s grant in 2019 amounted 400 000 rubles.
  • The total budget amounted 500 thousand rubles.
  • The funds were spent on completing the stated tasks and salaries for project employees.

Results: 

  1. The technique of layer-by-layer application of oppositely charged polymers was used. This technique reduced the rate of development of the amphipod immune response, but did not completely eliminate it.
  2. Immobilization of yeast in the strands of the hydrogel proved to be a more successful approach. It was found that the hydrogel does not cause the development of a cellular immune response for at least a few days, and also provides the ability to concentrate a large number of yeast cells while preserving their viability for a long time.
  3. Transgenic strain of yeast was derived to determine the concentration of copper. However, the simultaneous impact of factors such as low temperature, high concentration of yeast cells in the gel and low nutritional value of the components of the crustacean hemolymph led to the failure of this genetic system.
  4. Due to the reduced ability to synthesize protective proteins inside the gel, yeast shows increased sensitivity to copper. This effect made it possible to use as a functional component of microsensors a yeast strain that synthesizes a yellow fluorescent protein that quantitatively reacts to the appearance of copper in the medium.
  5. The invivo experiments allowed us to confirm the functionality of the developed biosensors for evaluating the increase in copper content in the hemolymph of Baikal endemic amphipods.

Conclusion of scientists:

The results obtained and the developed methods have significant potential for application and can be used in the future to develop effective procedures for environmental monitoring of lake Baikal communities.

Publications:

Application of PEG-Covered Non-Biodegradable Polyelectrolyte Microcapsules in the Crustacean Circulatory System on the Example of the Amphipod Eulimnogammarus verrucosus  — MPDI, 27.07.2019

Имплантируемые полиэлектролитные микросенсоры как инструмент для прижизненной динамической диагностики физиологического состояния ракообразных и оценка их влияния на организм – МАКС Пресс, 2019

«Об умных микролампочках в байкальских рачках» — Вести-Иркутск 9.06.2019

Три проекта молодых ученых получили гранты фонда «Озеро Байкал» — IRK.ru21.02.2019

Проекты по исследованию и спасению эндемиков поддержал фонд «Озеро Байкал» — Агентство Социальной информации – Иркутск 12.03.2019

Сюжет НИИ биологии ИГУ «Умные микролампочки в байкальских рачках 1/3: В сетях иммунной системы» 26.05.2019

Байкальская инициатива: будущее великого озера в руках молодых ученых – Экология и жизнь 20.02.2019



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