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Continuous transdermal monitoring of lactate in interstitial fluid (ISF) offers a minimally invasive approach for real\u2010time monitoring of metabolic status, particularly in conditions associated with metabolic syndrome such as obesity and diabetes. Here we report a proof\u2010of\u2010concept transdermal lactate biosensor based on a fenestrated microneedle array (F\u2010MNA), consisting of a polymeric MNA enclosed within a fenestrated polymeric shell that protects the biosensing layer while retaining access to ISF. Under controlled anesthetized conditions, the biosensor differentiated ISF lactate profiles between healthy and diabetic\u2010obese (db\/db) mice, with the db\/db group exhibiting higher baseline lactate concentrations. In fasted healthy mice subjected to subcutaneous glucose challenge, the biosensor captured transient increases in ISF lactate followed by partial recovery over the 2\u00a0h monitoring period. Biosensor\u2010derived ISF lactate showed strong temporal association with paired blood lactate measurements while exhibiting a systematic positive ISF\u2010to\u2010blood concentration offset. These findings establish proof\u2010of\u2010concept continuous transdermal ISF lactate monitoring under controlled in vivo conditions.<\/jats:p>","DOI":"10.1002\/smll.76250","type":"journal-article","created":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T13:25:25Z","timestamp":1791984325000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Fenestrated Microneedle Biosensor for Continuous In Vivo Lactate Monitoring in Interstitial Fluid"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9636-0718","authenticated-orcid":false,"given":"Masoud","family":"Khazaei","sequence":"first","affiliation":[{"name":"Monash Institute of Pharmaceutical Sciences Monash University  Parkville Victoria Australia"},{"name":"Victorian Node of the Australian National Fabrication Facility Melbourne Centre for Nanofabrication  Clayton Victoria 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