The ONI Blog

The best fluorophores for PALM imaging

Posted by Anna Caballe on 27 Jun 2019

Photoactivated Localization Microscopy, or PALM imaging, is a super-resolution technique that relies on the random activation of fluorophores to spatially resolve molecular details (Betzig et al. 2006). In brief, upon activation with the adequate laser, a sparse number of fluorophores emit for a short period before bleaching. Because fluorophores are only activated at the same time in small numbers until all have emitted, it is possible to localize and track single molecules over time.

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Topics: Technical, Microscopy

Using dSTORM to study purified extracellular vesicles

Posted by Andras Miklosi on 11 Jun 2019

Cells release different types of small-sized membrane vesicles (from 30nm to 1000nm) as a way of communicating with other cells and remove unwanted cell contents; these vesicles are collectively known as extracellular vesicles (EVs). Here we share how we have optimized the imaging of purified EVs to study their size, content and behavior using super-resolution microscopy.

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Topics: Technical, Microscopy

Our 9 tips to optimize your immunofluorescence staining

Posted by Anna Caballe on 15 May 2019

We have all been there, after spending a day or two fixing and staining your samples, the images don't look as good as expected, the signal is not optimal, and no obvious conclusion can be drawn from the data. We have put together some tips to help you optimize your immunofluorescence staining protocol and increase the signal-to-noise ratio in your samples. If you are unsure about how the technique works, we recommend you read our immunofluorescence guide first.

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Topics: Technical

Beginner's guide to STORM buffers

Posted by Mark Harmon on 28 Mar 2019

 When preparing a sample for STORM microscopy, one of the most important factors to consider is the choice of fluorophore. The ability to accurately localize single molecules is dependent on the use of fluorescent molecules that stochastically switch between a fluorescent “on” state and a dark “off” state – a photophysical behavior that is often referred to as blinking.

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Topics: Technical