Everspark
powered by the Eternity technology
Designed by Karine Monier, Arnaud Favier,
Christophe Place
"We designed Everspark buffer to facilitate sample preparation for dSTORM super-resolution imaging. Indeed, an oxygen-free buffer containing a thiol reducer is central to promote fluorophore blinking and thus single molecule detection, required for dSTORM imaging. With classical buffers, such as the Glox buffer, fluorophore blinking lifetime is limited to a few hours. With Everspark buffer, it is now possible to increase sample lifetime to several weeks. We demonstrated that a calibrated sample mounted once with the Everspark buffer can be submitted to several rounds of dSTORM imaging to acquire and reconstruct a structure reproducible in coverage and precision, over a period of two months."
EXAMPLES OF RESULTS
Everspark is a ready-to-use dSTORM super-resolution microscopy buffer offering long-term and stable fluorescence imaging over several weeks.
Two centrosomes imaged on a single slide, either the same Day (D0) and 7 days after mounting (D7) the sample with Eternity buffer. The typical 450 nm donut-like structure is visualised using a colour-coded scale encoded with the IGOR software, where each point appears as a function of its localisation precision (5 to 60 nm; inverted rainbow colour scale).
Labeling: Distal-appendages detected by immunofluorescence with AF647 in RPE-1 cells.
Right: The number of blinking events per centrosome and the median of the localisation precision in nm
Presented as a for each serie of 50,000 images recorded at D0 and D7 (left).
Credit: @Camille Fourneaux, Karine Monier - CRCL, Lyon
Each point is represented by its centroid (purple points) and its gaussian width (white). Intensity profil is displayed on the right with the measurement from peak to peak, in agreement with the size of the distal appendage crone.
Labeling: Distal-appendages detected by immunofluorescence with AF647 in RPE-1 cells.
Credit: @Corentin Rousset, Karine Monier, CRCL, Lyon
APPLICATIONS
TECHNOLOGY FEATURES
High stability over time: Samples prepared with Everspark are stable over several weeks
Wide pH range: Everspark buffer works over a wide pH range (5-8) where most biological phenomena occur, contrary to the enzyme-based buffer that requires a pH of 8 for efficient blinking
Efficiency: Everspark buffer provides highly efficient dark state transition
Multicolor: Everspark buffer is compatible with red and far red dSTORM blinking fluorophores (AF647, AF568, DL550, DL650)
METHODS
PUBLICATIONS