Max Planck Institute for Chemical Ecology, Max Planck Fellow Group On Plankton Community Interaction, Hans-Knöll-Str. 8, 07745, Jena, Germany.
Research Group Mass Spectrometry/Proteomics, Max Planck Institute for Chemical Ecology, Hans-Knöll-Str. 8, 07745, Jena, Germany.
Metabolomics. 2020 Feb 24;16(3):28. doi: 10.1007/s11306-020-1646-7.
Marine planktonic communities are complex microbial consortia often dominated by microscopic algae. The taxonomic identification of individual phytoplankton cells usually relies on their morphology and demands expert knowledge. Recently, a live single-cell mass spectrometry (LSC-MS) pipeline was developed to generate metabolic profiles of microalgae.
Taxonomic identification of diverse microalgal single cells from collection strains and plankton samples based on the metabolic fingerprints analyzed with matrix-free laser desorption/ionization high-resolution mass spectrometry.
Matrix-free atmospheric pressure laser-desorption ionization mass spectrometry was performed to acquire single-cell mass spectra from collection strains and prior identified environmental isolates. The computational identification of microalgal species was performed by spectral pattern matching (SPM). Three similarity scores and a bootstrap-derived confidence score were evaluated in terms of their classification performance. The effects of high and low-mass resolutions on the classification success were evaluated.
Several hundred single-cell mass spectra from nine genera and nine species of marine microalgae were obtained. SPM enabled the identification of single cells at the genus and species level with high accuracies. The receiver operating characteristic (ROC) curves indicated a good performance of the similarity measures but were outperformed by the bootstrap-derived confidence scores.
This is the first study to solve taxonomic identification of microalgae based on the metabolic fingerprints of the individual cell using an SPM approach.
海洋浮游生物群落是复杂的微生物群落，通常由微小的藻类主导。浮游植物细胞的分类鉴定通常依赖于其形态，需要专业知识。最近，开发了一种活单细胞质谱（LSC-MS）的方法来生成微藻的代谢图谱。
基于基质自由激光解吸/电离高分辨质谱分析的代谢指纹，对来自培养株和浮游生物样本的多种微藻单细胞进行分类鉴定。
对培养株和先前鉴定的环境分离株进行基质自由大气压激光解吸电离质谱，获取单细胞的质谱。通过光谱模式匹配（SPM）进行微藻种的计算鉴定。以分类性能为标准，评估了三种相似度得分和基于bootstrap 的置信度得分。评估了高和低质量分辨率对分类成功率的影响。
从 9 属 9 种海洋微藻中获得了几百个单细胞的质谱。SPM 可以实现对属和种水平单细胞的高准确度鉴定。接收者操作特性（ROC）曲线表明相似度度量的性能良好，但优于基于 bootstrap 的置信度得分。
这是第一项基于单细胞代谢指纹，使用 SPM 方法解决微藻分类鉴定的研究。
