%0 Journal Article %1 Lampe2012 %A Lampe, André %A Haucke, Volker %A Sigrist, Stephan J. %A Heilemann, Mike %A Schmoranzer, Jan %D 2012 %J Biology of the Cell %K fluorophores microscopy multicolor smlm spectral_demixing superresolution %N 4 %P 229--237 %R 10.1111/boc.201100011 %T Multi-colour direct STORM with red emitting carbocyanines %V 104 %X BACKGROUND INFORMATION: Single molecule-based super-resolution methods have become important tools to study nanoscale structures in cell biology. However, the complexity of multi-colour applications has prevented them from being widely used amongst biologists. Direct stochastic optical reconstruction microscopy (dSTORM) offers a simple way to perform single molecule super-resolution imaging without the need for an activator fluorophore and compatible with many conventionally used fluorophores. The search for the ideal dye pairs suitable for dual-colour dSTORM has been compromised by the fact that fluorophores spectrally apt for dual-colour imaging differ with respect to the optimal buffer conditions required for photoswitching and the generation of prolonged non-fluorescent (OFF) states.$\backslash$n$\backslash$nRESULTS: We present a novel variant of dSTORM that combines advantages of spectral demixing with the buffer compatible blinking properties of red emitting carbocyanine dyes, spectral demixing dSTORM (SD-dSTORM). In contrast to previously published work, SD-dSTORM requires reduced laser power and fewer imaging frames for the faithful reconstruction of super-resolved biological nanostructures. In addition, SD-dSTORM allows the use of commercially available rather than custom-made probes and does not rely on potentially error-prone cross-talk correction, thus allowing reliable co-localisation.$\backslash$n$\backslash$nCONCLUSIONS: SD-dSTORM presents a significant advance towards user-friendly single molecule localisation-based super-resolution microscopy combining advantages of state-of-the-art methodologies to perform fast, reliable and efficient multi-colour dSTORM. %@ 1768-322X (Electronic)$\backslash$r0248-4900 (Linking)

@article{Lampe2012, abstract = {BACKGROUND INFORMATION: Single molecule-based super-resolution methods have become important tools to study nanoscale structures in cell biology. However, the complexity of multi-colour applications has prevented them from being widely used amongst biologists. Direct stochastic optical reconstruction microscopy (dSTORM) offers a simple way to perform single molecule super-resolution imaging without the need for an activator fluorophore and compatible with many conventionally used fluorophores. The search for the ideal dye pairs suitable for dual-colour dSTORM has been compromised by the fact that fluorophores spectrally apt for dual-colour imaging differ with respect to the optimal buffer conditions required for photoswitching and the generation of prolonged non-fluorescent (OFF) states.$\backslash$n$\backslash$nRESULTS: We present a novel variant of dSTORM that combines advantages of spectral demixing with the buffer compatible blinking properties of red emitting carbocyanine dyes, spectral demixing dSTORM (SD-dSTORM). In contrast to previously published work, SD-dSTORM requires reduced laser power and fewer imaging frames for the faithful reconstruction of super-resolved biological nanostructures. In addition, SD-dSTORM allows the use of commercially available rather than custom-made probes and does not rely on potentially error-prone cross-talk correction, thus allowing reliable co-localisation.$\backslash$n$\backslash$nCONCLUSIONS: SD-dSTORM presents a significant advance towards user-friendly single molecule localisation-based super-resolution microscopy combining advantages of state-of-the-art methodologies to perform fast, reliable and efficient multi-colour dSTORM.}, added-at = {2020-03-23T21:12:34.000+0100}, author = {Lampe, Andr{\'{e}} and Haucke, Volker and Sigrist, Stephan J. and Heilemann, Mike and Schmoranzer, Jan}, biburl = {https://www.bibsonomy.org/bibtex/21c2f15b03642f9c5fb916e402fdf904c/kfriedl}, doi = {10.1111/boc.201100011}, file = {:C$\backslash$:/Users/Karoline/Documents/Strassburg{\_}NRP{\_}Projekt/Literature/Lampe (2012) Biol. Cell - spectra demixing dSTORM.pdf:pdf}, interhash = {6a1bcfeb1885cc6a7c8ba91e211f4663}, intrahash = {1c2f15b03642f9c5fb916e402fdf904c}, isbn = {1768-322X (Electronic)$\backslash$r0248-4900 (Linking)}, issn = {02484900}, journal = {Biology of the Cell}, keywords = {fluorophores microscopy multicolor smlm spectral_demixing superresolution}, number = 4, pages = {229--237}, pmid = {22187967}, timestamp = {2020-04-06T15:33:25.000+0200}, title = {{Multi-colour direct STORM with red emitting carbocyanines}}, volume = 104, year = 2012 }