Janelia Fluor® Dyes

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Janelia Fluor® 549 Conjugated Antibodies

Janelia Fluor® 646 Conjugated Antibodies

Janelia Fluor® Reactive Dyes

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Originally developed at the Janelia Research Campus, the Janelia Fluor® dyes have recently been made commercially available conjugated to antibodies or supplied as NHS esters for custom labeling. These bright, photostable dyes enable sensitive detection of cellular targets in IHC, ICC, and flow cytometry applications and are especially suited for live-cell imaging.


Tomm20 JF 646 Confocal Image JF 646 Excitation and Emission Spectra

Z-stack confocal images (top panel) with SRRF (super-resolution radial fluctuations) analysis (bottom) of Tomm20, an outer mitochondrial membrane protein, in HeLa cells. Tomm20 was detected in HeLa cells using 1 µg/ml Rb anti-Tomm20 (NBP1-81556) and 1:500 JF646-conjugated anti-Rb secondary antibody (red). Cells were counterstained with Dapi (blue). Scale bar: 10 μm. Experiment run by Jill Jenkins and images acquired at the Advanced Light Microscopy Core Facility, University of Colorado.

A single isolated Wistar rat cardiomyocyte stained against α-actinin, displaying its periodic structure localized at the ends of sarcomeres (Z-discs). Widefield fluorescence of JF549 (left panel), prior to illuminating with a 561 nm laser to induce photoswitching and produce a dSTORM super-resolution image (right). Scale bar: 1 μm. Images kindly provided by Prof. Christian Soeller, University of Exeter, acquired by Alex Clowsley and Anna Meletiou.



Find Janelia Fluor® conjugated antibodies from Novus and Janelia Fluor® reactive dyes from Tocris.



Benefits of the Janelia Fluor® dyes:

  • Bright and Photostable Dyes. Janelia Fluor® dyes have been specifically engineered to improve the brightness, fluorescence lifetime, and quantum yield of fluorescent dyes. Compared to Cy3 and TMR, Janelia Fluor® 549 is 2x brighter in vitro and in live-cell experiments.
  • Multicolor Detection and Analysis. The narrow emission spectra of Janelia Fluor® dyes enables combining multiple fluorescent labels to detect numerous cellular targets in a single tissue or cell sample.
  • Live-Cell Imaging of Intracellular Targets. Unlike most small organic fluorophores, Janelia Fluor® dyes are cell permeable. In addition, JF dyes are compatible with self-labeling systems such as HaloTag and SNAP-Tag, facilitating visualization and single molecule-tracking of intracellular targets in live cells.
  • Super Resolution Microscopy (SRM). Janelia Fluor® dyes are particularly useful for microscopy techniques including dSTORM (direct stochastic optical reconstruction microscopy) and STED (stimulated emission depletion).


Janelia Fluor® 549 (JF 549)

JF 549 Excitation and Emission Spectra
Emission Color Yellow
Brightness JF 549
Excitation max (nm) 549
Emission max (nm) 571
Extinction coefficient (ε, M-1cm-1) 101,000
Quantum yield (ф) 0.88
Common Equivalent Dyes Alexa Fluor 546, Alexa Fluor 555, Cy3, TRITC


Janelia Fluor® 646 (JF 646)

JF 646 Excitation and Emission Spectra
Emission Color Red
Brightness JF 646
Excitation max (nm) 646
Emission max (nm) 664
Max. extinction coefficient (ε, M-1cm-1) 152,000
Quantum yield (ф) 0.54
Common Equivalent Dyes Alexa Fluor 647, Atto 647, Cy5


SELECT CITATIONS OF JANELIA FLUOR DYES

Grimm, J.B., English, B.P., Chen. J., Slaughter, J.P., Zhang, Z., Revyakin, A., Patel, R., Macklin, J.J., Normanno, D., Singer, R.H., Lionnet, T., & Lavis, LD. (2015) A general method to improve fluorophores for live-cell and single-molecule microscopy. Nature Methods, 12,244. PMID:25599551

Grimm, J.B., English, B.P., Choi, H., Muthusamy, A.K., Mehl, B.P., Dong, P., Brown, T.A., Lippincott-Schwartz, J., Liu, Z., Lionnet, T., & Lavis, L.D. (2016) Bright photoactivatable fluorophores for single molecule imaging. Nature Methods, 13, 285. PMID 27776112

Grimm, J.B., Muthusamy, A.K., Liang, Y., Brown, T.A., Lemon, W.C., Patel, R., Lu, R., Macklin, J.J., Keller, P.J., Ji, N., & Lavis, L.D. (2017) A general method to fine-tune fluorophores for live-cell and in vivo imaging. Nature Methods, 14,987. PMID 28869757

Grimm, J.B., Brown, T.A., Tkachuk, A.N., & Lavis, L.D. (2017) A general method of Si-Fluoresceins and Si-Rhodamines. ACS Central Science, 3, 975. PMID:28979939

Lavis, L.D. (2017) Chemistry is Dead. Long Live Chemistry. Biochemistry, 56, 5165. PMID 28704030

Tsai, A., Muthusamy, A.K., Alves, M.R., Lavis, L.D., Singer, R.H., Stern, D.L., & Crocker, J. (2017) Nuclear microenvironments modulate transcription from low-affinity enhancers. Elife, 6, e28975. PMID: 29095143

Yadav, A.K., & Chan, J. (2017) Bright dyes bring biology into focus. ACS Central Science, 9, 920. PMID:28979930

Zhang, H., Aonbangkhen, C., Tarasovetc, E.V., Ballister, E.R., Chenoweth, D.M., & Lampson, M.A. (2017) Optogenetic control of kinetochore function. Nature Chemical Biology, 13, 1096. PMID: 28805800