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Here, we show that late embryogenesis abundant (LEA) proteins, previously shown to protect fragile samples from AWI-induced damage, can be leveraged to resolve sample orientation bias. The addition of LEA proteins can yield either new orientation distributions or more isotropic ones in samples suffering from orientation biases. These findings establish LEA proteins as a practical class of cryo-EM additives that can reduce AWI-induced damage and improve particle-orientation distributions in single-particle analysis.<\/jats:p>","DOI":"10.64898\/2026.07.09.737610","type":"posted-content","created":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T13:20:18Z","timestamp":1784208018000},"source":"Crossref","is-referenced-by-count":1,"title":["Diversifying particle-orientation distributions in cryo-EM with LEA protein additives"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6055-5604","authenticated-orcid":false,"given":"Kaitlyn M.","family":"Abe","sequence":"first","affiliation":[{"name":"Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy","family":"Grant","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA"},{"name":"John and Jeanne Rowe Center for Research in Virology, Morgridge Institute for Research, Madison, WI, 53715, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3327-1926","authenticated-orcid":false,"given":"Ci Ji","family":"Lim","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026071811550546000_2026.07.09.737610v1.1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1251652"},{"key":"2026071811550546000_2026.07.09.737610v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2023.102646"},{"key":"2026071811550546000_2026.07.09.737610v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/308032a0"},{"key":"2026071811550546000_2026.07.09.737610v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/0304-3991(82)90187-5"},{"key":"2026071811550546000_2026.07.09.737610v1.5","doi-asserted-by":"crossref","first-page":"50","DOI":"10.3389\/fmolb.2018.00050","article-title":"Benchmarking cryo-EM Single Particle Analysis Workflow","volume":"5","year":"2018","journal-title":"Front. 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