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It requires high-speed acquisition capabilities, low phototoxicity, and low mechanical disturbances. Three-dimensional imaging in microfluidic devices poses additional challenges as a deep penetration of the light source is required, along with a stationary setting, so the flows are not perturbed. Different types of fluorescence microscopy techniques have been used to address these limitations; particularly, confocal microscopy and light sheet fluorescence microscopy (LSFM). This manuscript proposes a novel architecture of a type of LSFM, single-plane illumination microscopy (SPIM). This custom-made microscope includes two mirror galvanometers to scan the sample vertically and reduce shadowing artifacts while avoiding unnecessary movement. In addition, two electro-tunable lenses fine-tune the focus position and reduce the scattering caused by the microfluidic devices. The microscope has been fully set up and characterized, achieving a resolution of 1.50 \u03bcm in the x-y plane and 7.93 \u03bcm in the z-direction. The proposed architecture has risen to the challenges posed when imaging microfluidic devices and live processes, as it can successfully acquire 3D volumetric images together with time-lapse recordings, and it is thus a suitable microscopic technique for live tracking miniaturized tissue and disease models.<\/jats:p>","DOI":"10.3390\/bios12121110","type":"journal-article","created":{"date-parts":[[2022,12,2]],"date-time":"2022-12-02T01:44:46Z","timestamp":1669945486000},"page":"1110","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Single Plane Illumination Microscopy for Microfluidic Device Imaging"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2426-9341","authenticated-orcid":false,"given":"Clara","family":"Gomez-Cruz","sequence":"first","affiliation":[{"name":"Departamento de Bioingenier\u00eda e Ingenier\u00eda Aeroespacial, Universidad Carlos III de Madrid, 28911 Legan\u00e9s, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3504-2051","authenticated-orcid":false,"given":"Sonia","family":"Laguna","sequence":"additional","affiliation":[{"name":"Departamento de Bioingenier\u00eda e Ingenier\u00eda Aeroespacial, Universidad Carlos III de Madrid, 28911 Legan\u00e9s, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ariadna","family":"Bachiller-Pulido","sequence":"additional","affiliation":[{"name":"Departamento de Bioingenier\u00eda e Ingenier\u00eda Aeroespacial, Universidad Carlos III de Madrid, 28911 Legan\u00e9s, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7707-8709","authenticated-orcid":false,"given":"Cristina","family":"Quilez","sequence":"additional","affiliation":[{"name":"Departamento de Bioingenier\u00eda e Ingenier\u00eda Aeroespacial, Universidad Carlos III de Madrid, 28911 Legan\u00e9s, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marina","family":"Ca\u00f1adas-Ortega","sequence":"additional","affiliation":[{"name":"Departamento de Bioingenier\u00eda e Ingenier\u00eda Aeroespacial, Universidad Carlos III de Madrid, 28911 Legan\u00e9s, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ignacio","family":"Albert-Smet","sequence":"additional","affiliation":[{"name":"Departamento de Bioingenier\u00eda e Ingenier\u00eda Aeroespacial, Universidad Carlos III de Madrid, 28911 Legan\u00e9s, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jorge","family":"Ripoll","sequence":"additional","affiliation":[{"name":"Departamento de Bioingenier\u00eda e Ingenier\u00eda Aeroespacial, Universidad Carlos III de Madrid, 28911 Legan\u00e9s, Spain"},{"name":"Instituto de Investigaci\u00f3n Sanitaria Gregorio Mara\u00f1\u00f3n, 28009 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1573-1661","authenticated-orcid":false,"given":"Arrate","family":"Mu\u00f1oz-Barrutia","sequence":"additional","affiliation":[{"name":"Departamento de Bioingenier\u00eda e Ingenier\u00eda Aeroespacial, Universidad Carlos III de Madrid, 28911 Legan\u00e9s, Spain"},{"name":"Instituto de Investigaci\u00f3n Sanitaria Gregorio Mara\u00f1\u00f3n, 28009 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1038\/nature05058","article-title":"The origins and the future of microfluidics","volume":"442","author":"Whitesides","year":"2006","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1583","DOI":"10.1039\/b820138b","article-title":"Single-layer planar on-chip flow cytometer using microfluidic drifting based three-dimensional (3D) hydrodynamic focusing","volume":"9","author":"Mao","year":"2009","journal-title":"Lab Chip"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1111\/j.1472-765X.2006.01953.x","article-title":"Application of a microfluidic device for counting of bacteria","volume":"43","author":"Inatomi","year":"2006","journal-title":"Lett. 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