Fluorescence microscopy is a defining technology in cellular biology but the diffraction of light limits the resolution of standard microscopes to ~200 nm, thus preventing detailed analyses of molecular structures. We are actively interested in microscopy methods based on stochastic photoswitching and computational localization of single fluorophores (“pointillism”), which enable greatly improved resolution. We have implemented pointillist super-resolution microscopy hardware and reconstruction software (Figure 2) and applied this imaging technique to various studies of pathogens in their cellular hosts [7][8][9][10].
In addition to pursuing biological applications, we aim to push some of the limitations of existing super-resolution methods. One such limitation is the use of invasive fluorescent tags, which can perturb biological functions. We demonstrated FlAsH-PALM, which combines FlAsH-tetracysteine tagging with stochastic localization microscopy. FlAsH-PALM allows to obtain <30 nm resolution images of delicate microbial proteins such as the HIV integrase, without disrupting their function [11][12].
We currently explore different experimental and computational strategies to further improve the information content, spatio-temporal resolution and reliability of pointillist microscopy.Figure 2: Super-resolution PALM/STORM imaging. Left: home-made imaging system. Middle: Dual color 3D STORM image of an axonal track, with a resolution of ~20 nm. Right: magnified view shows an the hollow structure of an individual ~50 nm diameter synaptic vesicle. From [13].
Fundings
[:en]We use cookies on our website to give you the most relevant experience by remembering your preferences and repeat visits. By clicking “Accept”, you consent to the use of ALL the cookies.[:fr]En cliquant sur « Accepter tous les cookies », vous acceptez le stockage de cookies sur votre appareil pour améliorer la navigation sur le site, analyser son utilisation et contribuer à nos efforts de marketing pour soutenir la recherche.[:] Read More
Cookies are small text files that can be used by websites to make a user's experience more efficient. The law states that we can store cookies on your device if they are strictly necessary for the operation of this site. For all other types of cookies we need your permission. This site uses different types of cookies. Some cookies are placed by third party services that appear on our pages.
Necessary cookies help make a website usable by enabling basic functions like page navigation and access to secure areas of the website. The website cannot function properly without these cookies.
Marketing cookies are used to track visitors across websites. The intention is to display ads that are relevant and engaging for the individual user and thereby more valuable for publishers and third party advertisers.
Preference cookies enable a website to remember information that changes the way the website behaves or looks, like your preferred language or the region that you are in.
![Figure 2: Super-resolution PALM/STORM imaging. Left: home-made imaging system. Middle: Dual color 3D STORM image of an axonal track, with a resolution of ~20 nm. Right: magnified view shows an the hollow structure of an individual ~50 nm diameter synaptic vesicle. From [13].](https://research.pasteur.fr/wp-content/uploads/2015/06/research.pasteur.fr_image-palm.png)
