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These H<jats:sub>2<\/jats:sub>O\u2010rich phase domains have been found to significantly influence material transportation and biochemical reactions. However, the accurate quantification of the size of these domains remains a challenge. Here, formulas are set up to calculate the anomalous diffusion exponent (<jats:italic>\u03b1<\/jats:italic>), the concentration threshold (<jats:italic>c<jats:sub>p<\/jats:sub><\/jats:italic>), and the radius of the H<jats:sub>2<\/jats:sub>O\u2010rich phase domain (<jats:italic>r<jats:sub>0<\/jats:sub><\/jats:italic>) to characterize the crowded solutions. Fitting coefficient (<jats:italic>R<jats:sup>2<\/jats:sup><\/jats:italic>) of the <jats:italic>r<jats:sub>0<\/jats:sub><\/jats:italic> fitted curves are 0.9989 for PEG\u20108k Da and 0.9901 for PEG\u201020k Da, respectively, which confirms the formulas to be suitable for quantifying the crowding degree. The values of <jats:italic>\u03b1<\/jats:italic>, <jats:italic>r<jats:sub>0<\/jats:sub><\/jats:italic>, and <jats:italic>c<jats:sub>p<\/jats:sub><\/jats:italic> of three different cell lysates is are calculated using these formulas. The <jats:italic>r<jats:sub>0<\/jats:sub><\/jats:italic> values of the cytosol from eukaryotic cells are 1.22\u00a0\u00b5m for HEK\u2010293T and 1.46\u00a0\u00b5m for <jats:italic>S. Cerevisiae<\/jats:italic>, respectively, which are smaller than that (2.13\u00a0\u00b5m) from prokaryotic cells (<jats:italic>E. coli<\/jats:italic>). This may be due to the more complex components, with higher molecular weight but lower concentration in the eukaryotic cells. This method for quantifying the H<jats:sub>2<\/jats:sub>O\u2010rich phase in a crowded solution helps to have a deeper understanding of the biochemical mechanism inside cells.<\/jats:p>","DOI":"10.1002\/smtd.202401905","type":"journal-article","created":{"date-parts":[[2025,2,17]],"date-time":"2025-02-17T15:20:56Z","timestamp":1739805656000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Elucidating Local Confinement in Crowded Polymer Solutions Within Giant Unilamellar Vesicles (GUVs) Through Single Particle Tracking Toward Deeper Understanding of Cells"],"prefix":"10.1002","volume":"9","author":[{"given":"Yingming","family":"Zhao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Urban Water Resource and Environment School of Chemistry and Chemical Engineering Harbin Institute of Technology  No. 92, West Da\u2010Zhi Street Harbin 150001 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shubin","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Urban Water Resource and Environment School of Chemistry and Chemical Engineering Harbin Institute of Technology  No. 92, West Da\u2010Zhi Street Harbin 150001 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanhao","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Urban Water Resource and Environment School of Chemistry and Chemical Engineering Harbin Institute of Technology  No. 92, West Da\u2010Zhi Street Harbin 150001 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chao","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Urban Water Resource and Environment School of Chemistry and Chemical Engineering Harbin Institute of Technology  No. 92, West Da\u2010Zhi Street Harbin 150001 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongshuo","family":"Ren","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Urban Water Resource and Environment School of Chemistry and Chemical Engineering Harbin Institute of Technology  No. 92, West Da\u2010Zhi Street Harbin 150001 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiangxiang","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Urban Water Resource and Environment School of Chemistry and Chemical Engineering Harbin Institute of Technology  No. 92, West Da\u2010Zhi Street Harbin 150001 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhao","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering The University of Hong Kong  Pokfulam 000000 Hong Kong"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mingdong","family":"Dong","sequence":"additional","affiliation":[{"name":"Interdisciplinary Nanoscience Center (iNANO) Aarhus University  Gustav Wieds Vej 14 (Building 1590), Gustav Wieds Vej Aarhu 8000 Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8571-6187","authenticated-orcid":false,"given":"Xiaojun","family":"Han","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Urban Water Resource and Environment School of Chemistry and Chemical Engineering Harbin Institute of Technology  No. 92, West Da\u2010Zhi Street Harbin 150001 China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,2,17]]},"reference":[{"key":"e_1_2_8_1_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms9352"},{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm.2017.7"},{"key":"e_1_2_8_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-05403-1"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1103\/PhysRevLett.96.098102"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0959-440X(00)00172-X"},{"key":"e_1_2_8_4_2","doi-asserted-by":"publisher","DOI":"10.3390\/membranes12060593"},{"key":"e_1_2_8_4_3","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(03)74856-5"},{"key":"e_1_2_8_4_4","doi-asserted-by":"publisher","DOI":"10.1088\/0034-4885\/76\/4\/046602"},{"key":"e_1_2_8_4_5","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.9b02811"},{"key":"e_1_2_8_4_6","doi-asserted-by":"publisher","DOI":"10.1021\/ma501608x"},{"key":"e_1_2_8_4_7","doi-asserted-by":"publisher","DOI":"10.1088\/1478-3975\/ab7f51"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nnano.2014.110"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nnano.2015.243"},{"key":"e_1_2_8_6_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1005122"},{"key":"e_1_2_8_6_3","first-page":"602","volume":"2013","author":"Tan C.","year":"2011","journal-title":"Nat. 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