On-Chip Photoacoustics-Activated Cell Sorting (PA-ACS) for Label-Free and High-Throughput Detection and Screening of Microalgal Cells,Analytical Chemistry

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On-Chip Photoacoustics-Activated Cell Sorting (PA-ACS) for Label-Free and High-Throughput Detection and Screening of Microalgal Cells
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-01-09 , DOI: 10.1021/acs.analchem.3c04665
Xiudong Duan ₁ , Xinqi Zheng ₁ , Ziyu Liu ₁ , Tianshu Dong ₁ , Yingdong Luo ₁ , Wei Yan ₂ , Cong Wang ₁ , Chaolong Song ₁

Affiliation

Microalgae play a crucial role in global carbon cycling as they convert carbon dioxide into various valuable macromolecules. Among them, Haematococcus pluvialis (H. pluvialis) is the richest natural source of astaxanthin (AXT), which is a valuable antioxidant, anti-inflammatory, and antiapoptosis agent. These benefits make AXT highly commercially valuable in pharmaceuticals, cosmetics, and nutritional industries. However, intrinsic genetic characteristics and extrinsic cultivation conditions influence biomass gains, leading to low productivity and extraction as the main techno-economic bottlenecks in this industry. Thus, detecting AXT in H. pluvialis is essential to determine the influence of multiple parameters on biocompound accumulation, enabling optimization of cultivation and enrichment of AXT-rich H. pluvialis cells. This work developed an opto-acousto-fluidic microplatform for detection, analysis, and sorting of microalgae. Via label-free monitoring and extraction of sample-induced ultrasonic signals, a photoacoustic microscopic system was proposed to provide a full-field visualization of AXT’s content and distribution inside H. pluvialis cells. When employed as on-chip image-based flow cytometry, our microplatform can also offer high-throughput measurements of intracellular AXT in real time, which demonstrates similar results to conventional spectrophotometry methods and further reveals the heterogeneity of AXT content at the single-cell level. In addition, a solenoid valve-pump dual-mode cell sorter was integrated for effective sorting of cells with a maximum working frequency of 0.77 Hz, reducing the fluid response time by 50% in rising and 40-fold in recovery. The H. pluvialis cells which have more AXT accumulation (>30 μm in diameter) were 4.38-fold enriched with almost no dead empty and small green cells. According to the results, automated and reliable photoacoustics-activated cell sorting (PA-ACS) can screen AXT-rich cells and remove impurities at the terminal stage of cultivation, thereby increasing the effectiveness and purity of AXT extraction. The proposed system can be further adopted to enrich strains and mutants for the production of biofuels or other rare organic substances such as β-carotene and lutein.

中文翻译：

用于微藻细胞无标记、高通量检测和筛选的片上光声激活细胞分选 (PA-ACS)

微藻将二氧化碳转化为各种有价值的大分子，在全球碳循环中发挥着至关重要的作用。其中，雨生红球藻（ H. pluvialis ）是虾青素（AXT）最丰富的天然来源，虾青素是一种有价值的抗氧化剂、抗炎剂和抗细胞凋亡剂。这些优点使 AXT 在制药、化妆品和营养品行业具有极高的商业价值。然而，内在遗传特征和外在栽培条件影响生物量增益，导致生产力低下和提取成为该行业的主要技术经济瓶颈。因此，检测雨生红细胞中的 AXT 对于确定多个参数对生物化合物积累的影响至关重要，从而能够优化富含 AXT 的雨生红细胞的培养和富集。这项工作开发了一个用于微藻检测、分析和分类的光声流体微平台。通过无标记监测和提取样品引起的超声波信号，提出了一种光声显微系统，以提供雨生红球菌细胞内 AXT 含量和分布的全场可视化。当用作基于图像的片上流式细胞术时，我们的微平台还可以实时提供细胞内 AXT 的高通量测量，这证明了与传统分光光度法类似的结果，并进一步揭示了单细胞水平上 AXT 含量的异质性。此外，集成电磁阀泵双模式细胞分选仪，可有效分选细胞，最高工作频率为0.77 Hz，使液体上升响应时间缩短50%，恢复时间缩短40倍。 H. 具有更多 AXT 积累（直径 >30 μm）的雨生红细胞富集 4.38 倍，几乎没有死亡的空细胞和小绿色细胞。根据结果，自动化且可靠的光声激活细胞分选（PA-ACS）可以筛选富含AXT的细胞并在培养末期去除杂质，从而提高AXT提取的有效性和纯度。所提出的系统可以进一步用于富集用于生产生物燃料或其他稀有有机物质（例如β-胡萝卜素和叶黄素）的菌株和突变体。

更新日期：2024-01-09

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