The Best Dyes for Super-Resolution Applications
Recent publications comparing synthetic dyes for super-resolution imaging have shown CF® dyes give the best performance for multiple methods. The superior brightness, photostability, and photochemical switching properties of certain CF® dyes are ideal for 3-D SIM, multi-color 3-D STORM, and other super-resolution and single molecule imaging techniques.
As Lehmann and colleagues report in their publication in the Journal of Biophotonics:
“…the spectrally close dyes CF®647 and CF®680 comprise an optimal dye pair for spectral demixing-based, registration free multicolor dSTORM with low crosstalk. Combining this dye pair with the separately excited CF®568 we performed 3-color dSTORM to image the relative nanoscale distribution of components of the endocytic machinery and the cytoskeleton.”
Single-Label Secondary Antibody Conjugates for STORM
Secondary antibodies with a low degree of labeling (DOL, or number of dye molecules per antibody molecule) have been reported to be optimal for STORM (Bittel et al. 2015). We now offer single-label secondary antibody conjugates with an average DOL of one for STORM applications.
Learn more about CF® Dyes for Super-Resolution Imaging:
References for CF® Dyes in Super-Resolution Microscopy
|Dye||Abs/Em maxima||Extinction coefficient||Super resolution application||References|
|CF®405S||404/431 nm||33,000||SIM||Demmerle et al. (2017). Strategic and practical guidelines for successful structured illumination microscopy. Nature Protocols 12, 988–1010.
Essig et al., (2017). Roquin Suppresses the PI3K-mTOR Signaling Pathway to Inhibit T Helper Cell Differentiation and Conversion
of Treg to Tfr Cells. Volume 47, Issue 6, p1067–1082.e12
|CF®405M||408/452 nm||41,000||SIM||Demmerle et al. (2017). Strategic and practical guidelines for successful structured illumination microscopy. Nature Protocols 12, 988–1010.
Kraus, F. et al. (2017) Quantitative 3D structured illumination microscopy of nuclear structures.Nat Protoc 12, 1011-1028, doi:nprot.2017.020 [pii]
Markaki, Y. et al. (2013). Fluorescence In Situ Hybridization Applications for Super-Resolution 3D Structured Illumination Microscopy. Methods Mol Biol 950, 43-64.
Miron, E. et. al. (2016). In Vivo and In Situ Replication Labeling Methods for Super-resolution Structured Illumination Microscopy of Chromosome Territories and Chromatin Domains. in Mark C. Leake (ed.), Chromosome Architecture: Methods and Protocols, Methods in Molecular Biology, vol. 1431, 127-140.
Ohgomori, T. et al. (2017). Differential activation of neuronal and glial STAT3 in the spinal cord of the SOD1G93A mouse model of amyotrophic lateral sclerosis.Eur J Neurosci 46, 2001-2014, doi:10.1111/ejn.13650
|CF®488A||490/515 nm||70,000||STED, TIRF||Angelov, B. & Angelova, A. (2017). Nanoscale clustering of the neurotrophin receptor TrkB revealed by super-resolution STED microscopy. Nanoscale 9, 9797-9804, doi:10.1039/c7nr03454g. (STED)
Zanetti-Domingues, L.C. et al. (2013). Hydrophobic Fluorescent Probes Introduce Artifacts into Single Molecule Tracking Experiments Due to Non-Specific Binding. PLoS ONE 8(9): e74200. (TIRF)
|CF®535ST||535/568 nm||95,000||STORM||Collaborator communication; for more information contact firstname.lastname@example.org.|
|CF®555||555/565 nm||150,000||Multicolor STORM||Lehmann, M. et al. (2015). Novel organic dyes for multicolor localization-based super-resolution microscopy.J Biophotonics DOI 10.1002/jbio.201500119|
|CF®568||562/583 nm||100,000||Multicolor STORM, SIM, TIRF||Gong, Y.-N. et al. (2017). Biological events and molecular signaling following MLKL activation during necroptosis. Cell Cycle, 1-13. doi:10.1080/15384101.2017.1371889 (STORM)|
Gorur, A. et al. (2017). COPII-coated membranes function as transport carriers of intracellular procollagen I. J Cell Biol 216, 1745-1759. doi:10.1083/jcb.201702135 (STORM)
Haas, K. et al. (2018). Single-molecule localization microscopy reveals molecular transactions during RAD51 filament assembly at cellular DNA damage sites. Nucleic Acids Research, Volume 46(5), 2398–2416. (STORM)
Heller, J. (2017). Exploring Nanoscale Organisation of Synapses with Super-Resolution Microscopy. OM&P 3, 48-58, doi:doi:10.20388/omp2017.002.0045 (STORM)
Jorgans, D.M. et al. (2017). Deep nuclear invaginations are linked to cytoskeletal filaments – integrated bioimaging of epithelial cells in 3D culture. J Cell Sci 2017 130: 177-189. doi: 10.1242/jcs.190967 (STORM)
Karanasios, E. et al. (2016). Autophagy initiation by ULK complex assembly on ER tubulovesicular regions marked by ATG9 vesicles.Nature Communications 7: 12420. DOI: 10.1038/ncomms12420 (STORM)
Kraus, F. et al. (2017) Quantitative 3D structured illumination microscopy of nuclear structures.Nat Protoc 12, 1011-1028, doi:nprot.2017.020 [pii] (SIM)
Lehmann, M. et al. (2015). Novel organic dyes for multicolor localization-based super-resolution microscopy.J Biophotonics DOI 10.1002/jbio.201500119 (STORM)
Lim, A. et al. (2017). Two kinesins drive anterograd