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After drift correction, 3D-DLS combined with SLS provides a consistent description of thermally induced collapse, pH-dependent swelling and core\u2013corona structure. Regarding drug delivery, loading efficiency for Doxorubicin and 5-Fluorouracil is maximized near the volume phase transition temperature, where hydrophobic interactions are strongest. For release studies, dialysis is recommended, but free-drug blanks are required to account for membrane-induced delay and ensure accurate early kinetic profiles. By integrating TEM, AFM, SLS, DLS, NTA and LDE, this study establishes a robust framework for the colloidal characterization of thermosensitive microgels. These refinements reduce experimental bias and may be extended to related soft nanocarriers.<\/jats:p>","DOI":"10.3390\/gels12070628","type":"journal-article","created":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T03:06:52Z","timestamp":1784084812000},"page":"628","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Protocol-Dependent Effects on Colloidal Characterization and Drug Loading\/Release Analysis of Thermosensitive PNIPAM-co-COOH Microgels"],"prefix":"10.3390","volume":"12","author":[{"given":"Jos\u00e9","family":"L\u00f3pez-Molina","sequence":"first","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-1033-7586","authenticated-orcid":false,"given":"Alba","family":"Garrido-Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6800-1489","authenticated-orcid":false,"given":"Mar\u00eda","family":"Tirado-Miranda","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Delfi","family":"Bastos-Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miguel","family":"Fern\u00e1ndez-Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmen","family":"Casas-Herce","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8043-0558","authenticated-orcid":false,"given":"Sol","family":"Esca\u00f1uela-Copado","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arturo","family":"Moncho-Jord\u00e1","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7997-8471","authenticated-orcid":false,"given":"Irene","family":"Adroher-Ben\u00edtez","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2422-655X","authenticated-orcid":false,"given":"J.","family":"L\u00f3pez-Romero","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, Faculty of Sciences, University of M\u00e1laga, 29071 M\u00e1laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6136-7477","authenticated-orcid":false,"given":"Ana","family":"J\u00f3dar-Reyes","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0869-9579","authenticated-orcid":false,"given":"Jos\u00e9","family":"Peula-Garc\u00eda","sequence":"additional","affiliation":[{"name":"Biocolloid and Fluid Physics Group, Applied Physics Department, University of Granada, Granada 18071, Spain"},{"name":"Department of Applied Physics II, University of M\u00e1laga, 29071 M\u00e1laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Thirupathi, K., Phan, T.T.V., Santhamoorthy, M., Ramkumar, V., and Kim, S.-C. 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