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Here, we introduce LUNA (Locking Under Nanoscale Accuracy), an innovative autofocusing method that leverages the coma pattern of detection light to characterize focus drift. LUNA improves the focusing precision down to 3 nm and extends the focusing range to at least 40 times the objective depth of focus. These advancements enable us to investigate the complete dynamics of bacterial single-cell CSR, revealing continuous cellular growth and division. We resolve a three-phase adaptation process characterized by distinct growth deceleration dynamics, and show that bacterial cells maintain robust size regulation and coordinate uniform adaptation to cold shock through synchronized growth and elapsed cycles. Notably, a model based on scattering theory reconciles the paradox between the growth lag of batch culture and continuous single-cell growth. These findings fundamentally transform our understanding of bacterial CSR and highlight LUNA\u2019s excellent potential for expanding state-of-the-art research in biology.<\/jats:p>","DOI":"10.7554\/elife.110268","type":"journal-article","created":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T17:30:15Z","timestamp":1773250215000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.110268","source":"Crossref","is-referenced-by-count":0,"title":["A coma pattern-based autofocusing method resolves bacterial cold shock response at single-cell level"],"prefix":"10.7554","volume":"15","author":[{"given":"Sihong","family":"Li","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04gh4er46","id-type":"ROR","asserted-by":"publisher"}],"name":"State Key Laboratory of Quantitative Synthetic Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences","place":["Shenzhen, 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