Odoo • Image et Texte

Everspark
 The ready-to-use dSTORM super-resolution microscopy buffer that offers long-term and stable fluorescence imaging over several weeks: pour it; seal it; image it!


Designed by Karine Monier, Arnaud Favier and Christophe Place

Reviewed by Pierre Bon, Ingrid Chamma, Benjamin Compans, Laurence Dubreil, Beatrice Durel, Tamas Gajdos, Sébastien Mailfert, Céline Malleval, Paolo Marchi, Delphine Muriaux, Jonny Nixon-Abell

Powered by the Eternity technology published in Scientific Reports

 

What is it for? Use it if you want someone to take care of the preparation of your blinking buffer or STED-derivative buffer, for MINFLUX-like techniques namely. Use it as well if you want to prepare samples on Monday, and image on Tuesday. Or even Friday!
Use it also if you want to prepare your samples in your lab, and image in your core facility
We prepare your buffer on demand for you. We deliver it when you need. Your buffer is ready-to-use: pour it, seal it, image it!
How to use it? Check our simplified protocol and the full protocol video
Why we chose it? Learn about the story of Everspark and us 
 Prices from 150€. See our offers.

TO THE SHOP

  Features

High stability over time

Samples are stable over several weeks

Wide pH range

Wide pH range (5-8) where most biological phenomena occur, contrary to enzyme-based buffers that require a pH of 8 for efficient blinking.

Efficiency

Highly efficient dark state transition

Multicolor

Compatible with red and far red dSTORM blinking fluorophores (AF647, AF568, DL550, DL650)

Ready to use

Can be used directly from the vial

2 months performance

Optimal performance for 2 months after production

Techniques

2D dSTORM
3D dSTORM
HILO, TIRF pattern.

Made on-demand

Fresh batches are produced at will, every week, for each order you make. They are delivered on the spot anywhere.

  Gallery

  The protocol in video

 
 
The recipe to prepare a good sealing
 
 

  Simplified protocol

Storage conditions: 

  • Everspark has to be stored in the fridge at 4°C upon receiving the product. Do not freeze it.

  • Open the tube just before mounting to limit oxygenation

  • Store the slides at 4°C and in the dark before repeating dSTORM acquisition on the same slide.

Use method:

  • Rinse the coverslip with sterile water

  • Discard water in excess

  • Prepare sealant: 1 volume of sealant catalyst + 1 volume of sealant basis in a micro-dish – do not mix

  • Open the Everspark tube and pipette right away the desired volume onto a Wilco glass bottom dish

  • Flip the coverslips face towards the slide on the cavity filled with Everspark without making bubbles

  • Absorb excess of Everspark buffer on the sides with a kimwipe

  • Mix sealant

  • Seal quickly with the reconstituted sealant

  • Wait for the sealant to polymerize on the slide before moving the slide

  • Discard the open tube of Everspark buffer (cannot be used for another slide, once already opened)

  • Gently clean the coverslip with 70% ethanol

  • Perform regular dSTORM acquisition (best to wait a couple of hours)

  • Repeat dSTORM acquisition 24 hours up to 3 weeks on the same slide providing no air bubbles were present

  • Store the slides at 4°C and in the dark before repeating dSTORM acquisition on the same slide (up to 3 weeks)

Note 1: Blinking events have been validated with Alexa-fluor 647, DL650, DL550 and CF568 (more to come)

Note 2: Blinking is detected for several weeks without loss of events or photons per event

Note 3: Mounting can also be performed in various microdishes by adjusting the Everspark volume (Each vial contains 450 µl)

BUY 5 VIALS AND TEST IT

Everspark and us

"We designed the 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."

Contact the team about their technology.

Odoo • Text and ImageKarine Monier - Christophe Place -  Arnaud Favier
Odoo • Image and Text Esther Graudens
New projects at Idylle

"When I was working on long manipulations, I dreamed of being able to stop at certain crucial stages. Everspark is the ideal answer to the problem of many researchers who prepare their samples in a hurry simply because they need to image them right away. The Everspark buffer eliminates this tension: they take the time to image whenever they want, with complete peace of mind. They can also make acquisitions over several days. This will really change the way people prepare their experimentations, and that's exactly what I like about my job!
In terms of production, I am very proud of the way we have managed to set up a just-in-time production process in our labs in record time. And when researchers ask us to introduce variants, our production teams are able to build and produce very quickly their customized buffer to test it".

Contact us about your sample preparation workflow

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