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Live imaging of Aiptasia larvae a model system for coral and anemone bleaching using a simple microfluidic device

机译:使用简单的微流控设备对Aiptasia幼虫进行实时成像这是一种用于珊瑚和海葵漂白的模型系统

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摘要

Coral reefs, and their associated diverse ecosystems, are of enormous ecological importance. In recent years, coral health has been severely impacted by environmental stressors brought on by human activity and climate change, threatening the extinction of several major reef ecosystems. Reef damage is mediated by a process called ‘coral bleaching’ where corals, sea anemones, and other cnidarians lose their photosynthetic algal symbionts (family Symbiodiniaceae) upon stress induction, resulting in drastically decreased host energy harvest and, ultimately, coral death. The mechanism by which this critical cnidarian-algal symbiosis is lost remains poorly understood. The larvae of the sea anemone, Exaiptasia pallida (commonly referred to as ‘Aiptasia’) are an attractive model organism to study this process, but they are large (∼100 mm in length, ∼75 mm in diameter), deformable, and highly motile, complicating long-term imaging and limiting study of this critical endosymbiotic relationship in live organisms. Here, we report ‘Traptasia’, a simple microfluidic device with multiple traps designed to isolate and image individual, live larvae of Aiptasia and their algal symbionts over extended time courses. Using a trap design parameterized via fluid flow simulations and polymer bead loading tests, we trapped Aiptasia larvae containing algal symbionts and demonstrated stable imaging for >10 hours. We visualized algae within Aiptasia larvae and observed algal expulsion under an environmental stressor. To our knowledge, this device is the first to enable time-lapsed, high-throughput live imaging of cnidarian larvae and their algal symbionts and, in further implementation, could provide important insights into the cellular mechanisms of cnidarian bleaching under different environmental stressors. The ‘Traptasia’ device is simple to use, requires minimal external equipment and no specialized training to operate, and can easily be adapted using the trap optimization data presented here to study a variety of large, motile organisms.
机译:珊瑚礁及其相关的多样化生态系统具有巨大的生态重要性。近年来,人类活动和气候变化给环境带来的压力极大地影响了珊瑚的健康,威胁到几个主要珊瑚礁生态系统的灭绝。珊瑚礁的破坏是通过称为“珊瑚白化”的过程来进行的,在该过程中,珊瑚,海葵和其他刺胞在压力诱导下失去了光合作用的藻类共生体(共生双歧杆菌科),导致宿主能量的大量减少,最终导致珊瑚死亡。这种关键的刺鼻-藻类共生丧失的机制仍然知之甚少。海葵幼虫,Exaiptasia pallida(通常称为“ Aiptasia”)是研究此过程的一种引人注目的模型生物,但它们很大(长约100毫米,直径约75毫米),可变形且高度变形运动,使长期成像变得复杂,并限制了在活生物体中这种关键的内共生关系的研究。在这里,我们报告“ Traptasia”,这是一种带有多个陷阱的简单微流控设备,旨在在较长的时间范围内分离和成像Aiptasia的单个活幼虫及其藻类共生体。使用通过流体流动模拟和聚合物珠负载测试参数化的疏水阀设计,我们捕获了包含藻类共生体的拟南芥幼虫,并显示了稳定的成像时间超过10小时。我们可视化了Aiptasia幼虫内的藻类,并观察到在环境胁迫下藻类的排出。据我们所知,该设备是第一个能够对cnidarian幼虫及其藻类共生体进行延时,高通量实时成像的设备,并且在进一步实施中,可以为了解不同环境压力下cnidarian漂白的细胞机制提供重要见解。 “ Traptasia”设备易于使用,所需外部设备最少,无需经过专门培训即可操作,并且可以使用此处提供的陷阱优化数据轻松地进行调整,以研究各种大型活动生物。

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