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Particularly in microfluidic cultivation and screening, precisely controlled environmental conditions and defined minimal media become important for quantitative studies. Nevertheless, conventional bulk media optimized for other scales, are often employed despite differences in nutrient requirements across varying scales and cultivation modes. This study demonstrates the limitations of using bulk media for single\u2010cell analysis of\n                    <jats:italic>Corynebacterium glutamicum<\/jats:italic>\n                    and highlights the need for revised minimal media for microfluidic cultivations under perfusion. The commonly used CGXII medium contains protocatechuic acid (PCA), which functions both as an iron chelator and a carbon source, thereby compromising its role as a minimal medium with one limiting substrate. We systematically evaluated alternative iron chelators to replace PCA using microfluidic single\u2010cell cultivation. We show that not all chelators are applicable with\n                    <jats:italic>C. glutamicum<\/jats:italic>\n                    at the single\u2010cell level and identify citrate as a suitable replacement that ensures iron availability without supporting growth in the absence of glucose. Based on these findings, we established a revised minimal medium, termed \u2018single\u2010cell CGXII\u2019. This medium was validated across different scales and enables robust, reproducible investigation of cellular physiology at single\u2010cell resolution, improving accuracy and comparability in quantitative single\u2010cell research.\n                  <\/jats:p>","DOI":"10.1002\/bit.70203","type":"journal-article","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T04:23:41Z","timestamp":1776227021000},"page":"1648-1660","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment of a Single\u2010Cell Minimal Medium: A Case Study for Microfluidic Cultivation of\n                    <i>Corynebacterium glutamicum<\/i>"],"prefix":"10.1002","volume":"123","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-1412-946X","authenticated-orcid":false,"given":"Judith","family":"Wegmann","sequence":"first","affiliation":[{"name":"Institute of Process Engineering in Life Sciences, Karlsruhe Institute of Technology 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