title: Brilliant violet fluorophores: a new class of ultrabright fluorescent compounds for immunofluorescence experiments.
abstract: The Nobel Prize in Chemistry was awarded in 2000 for the discovery of conductive organic polymers, which have subsequently been adapted for applications in ultrasensitive biological detection. Here, we report the first use of this new class of fluorescent probes in a diverse range of cytometric and imaging applications. We demonstrate that these "Brilliant Violet" reporters are dramatically brighter than other UV-violet excitable dyes, and are of similar utility to phycoerythrin (PE) and allophycocyanin (APC). They are thus ideally suited for cytometric assays requiring high sensitivity, such as MHC-multimer staining or detection of intracellular antigens. Furthermore, these reporters are sensitive and spectrally distinct options for fluorescence imaging, two-photon microscopy and imaging cytometry. These ultra-bright materials provide the first new high-sensitivity fluorescence probes in over 25 years and will have a dramatic impact on the design and implementation of multicolor panels for high-sensitivity immunofluorescence assays.
authors.affiliation: ImmunoTechnology Section, Vaccine Research Center, NIAID, NIH, Bethesda, Maryland 20892, USA. pchattop@mail.nih.gov
authors.forename: Pratip K
authors.initials: PK
authors.lastname: Chattopadhyay
authors.forename: Brent
authors.initials: B
authors.lastname: Gaylord
authors.forename: Adrian
authors.initials: A
authors.lastname: Palmer
authors.forename: Nan
authors.initials: N
authors.lastname: Jiang
authors.forename: Mary A
authors.initials: MA
authors.lastname: Raven
authors.forename: Geoff
authors.initials: G
authors.lastname: Lewis
authors.forename: Morgan A
authors.initials: MA
authors.lastname: Reuter
authors.forename: A K M
authors.initials: AK
authors.lastname: Nur-ur Rahman
authors.forename: David A
authors.initials: DA
authors.lastname: Price
authors.forename: Michael R
authors.initials: MR
authors.lastname: Betts
authors.forename: Mario
authors.initials: M
authors.lastname: Roederer
mesh_terms: D000818:Animals; D001921:Brain / Q000648:ultrastructure; D004553:Electric Conductivity; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton; D057054:Molecular Imaging; D010798:Phycocyanin / Q000032:analysis; D010799:Phycoerythrin / Q000032:analysis; D011108:Polymers / Q000032:analysis* / Q000737:chemistry; D012160:Retina / Q000648:ultrastructure; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D011108:Polymers; C002497:allophycocyanin; D010798:Phycocyanin; D010799:Phycoerythrin