Matches in SemOpenAlex for { <https://semopenalex.org/work/W4310258517> ?p ?o ?g. }
- W4310258517 abstract "Fixing cells with paraformaldehyde (PFA) is an essential step in numerous biological techniques as it is thought to preserve a snapshot of biomolecular transactions in living cells. Fixed-cell imaging techniques such as immunofluorescence have been widely used to detect liquid–liquid phase separation (LLPS) in vivo. Here, we compared images, before and after fixation, of cells expressing intrinsically disordered proteins that are able to undergo LLPS. Surprisingly, we found that PFA fixation can both enhance and diminish putative LLPS behaviors. For specific proteins, fixation can even cause their droplet-like puncta to artificially appear in cells that do not have any detectable puncta in the live condition. Fixing cells in the presence of glycine, a molecule that modulates fixation rates, can reverse the fixation effect from enhancing to diminishing LLPS appearance. We further established a kinetic model of fixation in the context of dynamic protein–protein interactions. Simulations based on the model suggest that protein localization in fixed cells depends on an intricate balance of protein–protein interaction dynamics, the overall rate of fixation, and notably, the difference between fixation rates of different proteins. Consistent with simulations, live-cell single-molecule imaging experiments showed that a fast overall rate of fixation relative to protein–protein interaction dynamics can minimize fixation artifacts. Our work reveals that PFA fixation changes the appearance of LLPS from living cells, presents a caveat in studying LLPS using fixation-based methods, and suggests a mechanism underlying the fixation artifact." @default.
- W4310258517 created "2022-11-30" @default.
- W4310258517 creator A5022938293 @default.
- W4310258517 creator A5058180819 @default.
- W4310258517 creator A5067345132 @default.
- W4310258517 creator A5091481135 @default.
- W4310258517 date "2022-11-29" @default.
- W4310258517 modified "2023-10-14" @default.
- W4310258517 title "Fixation can change the appearance of phase separation in living cells" @default.
- W4310258517 cites W1826402150 @default.
- W4310258517 cites W1828400526 @default.
- W4310258517 cites W1963175326 @default.
- W4310258517 cites W1974117072 @default.
- W4310258517 cites W1992858116 @default.
- W4310258517 cites W1995311028 @default.
- W4310258517 cites W2001858363 @default.
- W4310258517 cites W2010519082 @default.
- W4310258517 cites W2010959934 @default.
- W4310258517 cites W2017101979 @default.
- W4310258517 cites W2019374755 @default.
- W4310258517 cites W2027269991 @default.
- W4310258517 cites W2033265108 @default.
- W4310258517 cites W2048608623 @default.
- W4310258517 cites W2049869291 @default.
- W4310258517 cites W2059699217 @default.
- W4310258517 cites W2073026107 @default.
- W4310258517 cites W2085978470 @default.
- W4310258517 cites W2089717004 @default.
- W4310258517 cites W2096275862 @default.
- W4310258517 cites W2098669559 @default.
- W4310258517 cites W2102653059 @default.
- W4310258517 cites W2105311961 @default.
- W4310258517 cites W2129726480 @default.
- W4310258517 cites W2138503623 @default.
- W4310258517 cites W2139066576 @default.
- W4310258517 cites W2147076374 @default.
- W4310258517 cites W2150313376 @default.
- W4310258517 cites W2167279371 @default.
- W4310258517 cites W2168639701 @default.
- W4310258517 cites W2222857089 @default.
- W4310258517 cites W2271817176 @default.
- W4310258517 cites W2402201377 @default.
- W4310258517 cites W2406725268 @default.
- W4310258517 cites W2492265454 @default.
- W4310258517 cites W2507000097 @default.
- W4310258517 cites W2538965655 @default.
- W4310258517 cites W2567008309 @default.
- W4310258517 cites W2590568317 @default.
- W4310258517 cites W2603007067 @default.
- W4310258517 cites W2615607456 @default.
- W4310258517 cites W2729283896 @default.
- W4310258517 cites W2756821926 @default.
- W4310258517 cites W2770336503 @default.
- W4310258517 cites W2774429523 @default.
- W4310258517 cites W2781628200 @default.
- W4310258517 cites W2794654572 @default.
- W4310258517 cites W2796769720 @default.
- W4310258517 cites W2808847099 @default.
- W4310258517 cites W2810866092 @default.
- W4310258517 cites W2901209536 @default.
- W4310258517 cites W2902026154 @default.
- W4310258517 cites W2914907359 @default.
- W4310258517 cites W2942748350 @default.
- W4310258517 cites W2966567728 @default.
- W4310258517 cites W2979601486 @default.
- W4310258517 cites W2989281092 @default.
- W4310258517 cites W3016592635 @default.
- W4310258517 cites W3023602638 @default.
- W4310258517 cites W3043530209 @default.
- W4310258517 cites W3092217728 @default.
- W4310258517 cites W3117500204 @default.
- W4310258517 cites W3127108684 @default.
- W4310258517 cites W3158765559 @default.
- W4310258517 cites W3166144248 @default.
- W4310258517 cites W3190580055 @default.
- W4310258517 cites W3206740769 @default.
- W4310258517 cites W3209494177 @default.
- W4310258517 cites W3213943866 @default.
- W4310258517 cites W4206315000 @default.
- W4310258517 cites W4211227438 @default.
- W4310258517 cites W4221137519 @default.
- W4310258517 cites W4226293713 @default.
- W4310258517 cites W4229375645 @default.
- W4310258517 cites W4293576390 @default.
- W4310258517 cites W4305014650 @default.
- W4310258517 cites W793787745 @default.
- W4310258517 doi "https://doi.org/10.7554/elife.79903" @default.
- W4310258517 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/36444977" @default.
- W4310258517 hasPublicationYear "2022" @default.
- W4310258517 type Work @default.
- W4310258517 citedByCount "24" @default.
- W4310258517 countsByYear W43102585172022 @default.
- W4310258517 countsByYear W43102585172023 @default.
- W4310258517 crossrefType "journal-article" @default.
- W4310258517 hasAuthorship W4310258517A5022938293 @default.
- W4310258517 hasAuthorship W4310258517A5058180819 @default.
- W4310258517 hasAuthorship W4310258517A5067345132 @default.
- W4310258517 hasAuthorship W4310258517A5091481135 @default.
- W4310258517 hasBestOaLocation W43102585171 @default.
- W4310258517 hasConcept C104317684 @default.