Quantitative super-resolution localization microscopy of DNA in situ using Vybrant® DyeCycle™ Violet fluorescent probe

详细信息    查看全文

• 作者：Dominika Żurek-Biesiada^a ; Aleksander T. Szczurek^b ; Kirti Prakash^b ; ^c ; Gerrit Best^d ; Giriram K. Mohana^b ; Hyun-Keun Lee^b ; ^e ; Jean-Yves Roignant^b ; Jurek W. Dobrucki^a ; ^{jerzy.dobrucki@uj.edu.pl" class="auth_mail" title="E-mail the corresponding author} ; Christoph Cremer^b ; ^c ; ^d ; ^e ; ^{c.cremer@imb-mainz.de" class="auth_mail" title="E-mail the corresponding author} ; Udo Birk^b ; ^d ; ^e
• 关键词：Super-Resolution ; DNA ; dSTORM ; Localization microscopy ; Fluorescence ; Chromatin ; Vybrant violet ; DNA dye ; Single molecules ; Nucleus
• 刊名：Data in Brief
• 出版年：2016
• 出版时间：June 2016
• 年：2016
• 卷：7
• 期：Complete
• 页码：157-171
• 全文大小：5564 K

文摘

Single Molecule Localization Microscopy (SMLM) is a recently emerged optical imaging method that was shown to achieve a resolution in the order of tens of nanometers in intact cells. Novel high resolution imaging methods might be crucial for understanding of how the chromatin, a complex of DNA and proteins, is arranged in the eukaryotic cell nucleus. Such an approach utilizing switching of a fluorescent, DNA-binding dye Vybrant® DyeCycle™ Violet has been previously demonstrated by us (Żurek-Biesiada et al., 2015) [1]. Here we provide quantitative information on the influence of the chemical environment on the behavior of the dye, discuss the variability in the DNA-associated signal density, and demonstrate direct proof of enhanced structural resolution. Furthermore, we compare different visualization approaches. Finally, we describe various opportunities of multicolor DNA/SMLM imaging in eukaryotic cell nuclei.