Dataset for "A Microfluidic Spore Chamber for Long-Term Imaging of Single-Spore Hyphal Development"

Understanding the life cycle of fungal spores is essential for elucidating their roles in pathogenesis, dispersal, and survival. However, studying spore development under controlled, spatially defined conditions remains challenging. Here, we present the Spore Chamber, a custom-built microfluidic platform engineered for parallel trapping and long-term imaging of individual spores under defined media conditions, enabling real-time visualization of hyphal development. Using Aspergillus fumigatus as a model organism, we demonstrate that sparse trapping of individual spores within size-matched trap geometries enables long-term time-lapse imaging of key developmental stages, including germination, polarized hyphal elongation, branching, and conidiophore formation. To assess the device’s capacity to resolve morphogenetic responses to exogenous signals, we introduced lipochitooligosaccharides (LCOs) and short-chain chitooligosaccharides (COs). Rhizobium-derived, non-sulfated LCO (nsLCO) mixtures induced enhanced secondary branching (hyperbranching), a response not previously reported in A. fumigatus under these signal conditions, to our knowledge, whereas sulfated LCOs and CO4 did not significantly alter branching patterns. In addition, long-term confinement and imaging revealed rare developmental morphologies previously described primarily in mutant strains, including split conidiophore formation, elongated phialides, microcyclic conidiation, and chlamydospore development. Together, these results establish the Spore Chamber as a targeted microfluidic platform for single-spore phenotyping and long-term developmental analysis, with applications in fungal biology, chemical signaling studies, and host–microbe interaction research. Videos of the observed phenomena are included in this data set.

真菌孢子在孢子传播、菌体存活与致病性调控中发挥关键作用，然而以单细胞分辨率解析其发育过程仍存在诸多挑战。本研究介绍了弹珠式孢子培养腔（Pachinko Spore Chamber）：一款高通量微流控平台，可实现单个真菌孢子的捕获、空间隔离与长期动态监测。该装置可支持真菌发育过程的实时成像，能够精准量化孢子萌发、菌丝生长及分生孢子梗形成等关键过程，同时最大限度降低孢子间的相互干扰。本研究以烟曲霉（Aspergillus fumigatus）为实验对象，不仅观测到了预期的发育转变过程，还发现了此前未被报道或极为罕见的表型，包括厚垣孢子形成、分生孢子梗分裂、加长型分生孢子链以及微循环产孢，这些表型或受微环境限制的影响。本研究证实弹珠式孢子培养腔可作为一款通用且可扩展的研究平台，用于真菌表型分析、抗真菌药物筛选以及胁迫响应研究，为受控条件下真菌发育与存活能力的解析提供了全新视角。本数据集收录了上述观测现象的相关视频。