Vuille-Dit-Bille Emilie, Fonta Céline Loussert, Heub Sarah, Boder-Pasche Stéphanie, Sakar Mahmut Selman, Weder Gilles
CSEM SA, Neuchâtel, Switzerland.
Institute of Mechanical Engineering, EPFL, Lausanne, Switzerland.
Lab Chip. 2025 May 6. doi: 10.1039/d5lc00153f.
Miniaturized three-dimensional (3D) cell culture systems, in particular organoids and spheroids, hold great potential for studying morphogenesis, disease modeling, and drug discovery. However, sub-cellular resolution 3D imaging of these biological samples remains a challenge. Histology, the gold standard for microscopic interrogation of tissue anatomy, may address this challenge once the associated techniques are adapted. Due to their small size and delicate structure, organoids must be embedded in a supporting hydrogel. The histological sections have low information content because the distribution of the organoids within the gel is not controlled. To address this issue, we introduce an acoustic micromanipulation platform that concentrates and aligns organoids within a histology-compatible hydrogel block. Utilizing an array of micromachined lead zirconate titanate (PZT) transducers, the platform generates localised and precisely controlled acoustic standing waves to levitate organoids to a prescribed plane and fix their positions within a polyethylene glycol diacrylate (PEGDA)-gelatine hydrogel. Organoids from different culture conditions can be co-embedded in a traceable fashion with the use of a custom-design hydrogel grid. Our results demonstrate that more than 70% of spheroids can be positioned within a 150 μm-thick hydrogel block, substantially increasing the information content of histology sections. The platform's versatility, scalability, and ease of use will make histological assessment accessible to every life science laboratory.
小型化三维（3D）细胞培养系统，特别是类器官和球体，在研究形态发生、疾病建模和药物发现方面具有巨大潜力。然而，对这些生物样本进行亚细胞分辨率的3D成像仍然是一项挑战。组织学作为对组织解剖结构进行微观检查的金标准，一旦相关技术得到改进，可能会解决这一挑战。由于类器官体积小且结构脆弱，必须将其嵌入支撑水凝胶中。由于凝胶中类器官的分布不受控制，组织学切片的信息含量较低。为了解决这个问题，我们引入了一个声学微操纵平台，该平台可以在与组织学兼容的水凝胶块内聚集并排列类器官。利用一系列微加工的锆钛酸铅（PZT）换能器，该平台产生局部且精确控制的声学驻波，将类器官悬浮到规定平面，并将它们固定在聚乙二醇二丙烯酸酯（PEGDA）-明胶水凝胶内的位置。使用定制设计的水凝胶网格，可以以可追踪的方式将来自不同培养条件的类器官共嵌入。我们的结果表明，超过70%的球体可以定位在150μm厚的水凝胶块内，大大增加了组织学切片的信息含量。该平台的多功能性、可扩展性和易用性将使每个生命科学实验室都能进行组织学评估。
