arXiv:2506.03738physics.bio-phcs.SY2025-06被引 7

通过全息流式显微技术实现微藻三维形态与折射率重建,助力海洋生态监测。

3D Holographic Flow Cytometry Measurements of Microalgae: Strategies for Angle Recovery in Complex Rotation Patterns

  • 针对不同旋转模式设计角度恢复策略,提升3D重构精度。
  • 对透明低散射微藻可实现定量折射率映射,完整表征内外结构。
  • 即使在强吸收或高散射条件下,仍能完成硅藻和甲藻的3D形状重建。

海洋生态系统面临环境变化威胁,微藻在浮游与底栖生态中扮演关键角色,是全球监测的重要目标。然而,其空间与时间分布高度变异,物种鉴定依赖显微观察,且缺乏有效的3D形态分析技术。本文提出一套全流程方法,基于流动式全息显微术获取具有复杂几何形态的微藻3D信息。根据样本旋转模式采用定制化角度恢复策略。结果表明,对准透明、低散射的微生物,可实现定量3D折射率(RI)断层成像,完整揭示其内部结构与外部形态。即便在最复杂场景下,如高光吸收或强散射,仍可实现硅藻与甲藻的定量3D形状重建。对比商用系统通过2D推断的3D特性,本方法实现非侵入式3D测量,为水生生态系统研究提供全新生物洞察。

原文摘要 · Abstract (English)

Marine ecosystems are in the spotlight, because environmental changes are threatening biodiversity and ecological functions. In this context, microalgae play key ecological roles both in planktonic and benthic ecosystems. Consequently, they are considered indispensable targets for global monitoring programs. However, due to a high spatial and temporal variability and to difficulties of species identification (still relying on microscopy observations), the assessment of roles played by these components of marine ecosystems is demanding. In addition, technologies for a 3D assessment of their complex morphology are scarcely available. Here, we present a comprehensive workflow for retrieving 3D information on microalgae with diverse geometries through holographic microscopy operating in flow-cytometry mode. Depending on the rotation patterns of samples, a tailored approach is used to retrieve their rolling angles. We demonstrate the feasibility of measuring 3D data of various microalgae, contingent to the intrinsic optical properties of cells. Specifically, we show that for quasi-transparent and low-scattering microorganisms, the retrieved angles permit to achieve quantitative 3D tomographic Refractive Index (RI) mapping, providing a full characterization of the alga in terms of its inner structure and the outer shape. Moreover, even in the most challenging scenarios, where microalgae exhibit high light absorption or strong scattering, quantitative 3D shape reconstructions of diatoms and dinoflagellates can be at least achieved. Finally, we compare our direct 3D measurements with 2D inferences of 3D properties, obtained using a commercially available microscopy system. The ability to non-invasively obtain 3D information on microalgae marks a fundamental advancement in the field, unlocking a wealth of novel biological insights for characterizing aquatic ecosystems.

全息显微微藻分析3D重建海洋生态

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