Abstract
Super Resolution (SR) microscopy has become a powerful tool to study cellular architecture at the nanometer scale. Single molecule localization microscopy (SMLM) is a method in which fluorophore labels repeatedly switch On and Off (“blink”). Their exact locations are estimated by computing the centers of individual blinks. Therefore, the image quality depends on the density of the detected labels, as well as the accuracy of the estimation of their location. Both are influenced by several factors. Here we present a step-by-step method that optimizes many of these factors to facilitate multicolor imaging.
The authors Leila Nahidiazar and Rolf Harkes contributed equally.
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References
Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess HF (2006) Science 313(5793):1642. https://doi.org/10.1126/science.1127344
Hess ST, Girirajan TP, Mason MD (2006) Biophys J 91(11): 4258. https://doi.org/10.1529/biophysj.106.091116
Rust MJ, Bates M, Zhuang X (2006) Nature Methods 3(10):793. https://doi.org/10.1038/nmeth929
Heilemann M., van de Linde S, Schüttpelz M, Kasper R, Seefeldt B, Mukherjee A, Tinnefeld P, Sauer M (2008) Angew Chem Int Ed 47(33):6172. https://doi.org/10.1002/anie.200802376
Dempsey GT, Vaughan JC, Chen KH, Bates M, Zhuang X (2011) Nature Methods 8(12):1027 (2011). https://doi.org/10.1038/nmeth.1768
Nahidiazar L, Agronskaia AV, Broertjes J, van den Broek B, Jalink K (2016) PLOS ONE 11(7):e0158884. https://doi.org/10.1371/journal.pone.0158884
Nahidiazar L, Kreft M, van den Broek B, Secades P, Manders EMM, Sonnenberg A, Jalink K (2015) J Cell Sci 128(20):3714. https://doi.org/10.1242/jcs.171892
Simons CAJ, Lam HT (1977) Apparatus for reading an optically readable reflecting information structure
Ovesný M, Křížek P, Borkovec J, Švindrych Z, Hagen GM (2014) Bioinformatics 30(16):2389. https://doi.org/10.1093/bioinformatics/btu202
Rueden CT, Schindelin J, Hiner MC, DeZonia BE, Walter AE, Arena ET, Eliceiri KW (2017) BMC Bioinformatics 18(1):1–26. https://doi.org/10.1186/s12859-017-1934-z
Hoogendoorn E, Crosby KC, Leyton-Puig D, Breedijk RMP, Jalink K, Gadella TWJ, Postma M (2017) Sci Rep 4(1). https://doi.org/10.1038/srep03854
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Nahidiazar, L., Harkes, R. (2021). Multicolor Localization-Based Super Resolution Microscopy. In: Zamir, E. (eds) Multiplexed Imaging. Methods in Molecular Biology, vol 2350. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1593-5_5
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DOI: https://doi.org/10.1007/978-1-0716-1593-5_5
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