|Karolinska Institutet - Department of Medical Biochemistry and Biophysics
|Bidrag från Vinnova
|1 617 864 kronor
|juli 2017 - juni 2019
Syfte och mål
The goal of the project is to connect morphological and genetic information on the single cell level, using DNA nanostructures as labels. The nanostructures carry a set of sequences which make them detectable as a sequence of five color sites in fluorescent microscopy, using exchange DNA PAINT. The set of about a hundred of these structures are attached to cells which are then sequenced, during which another sequence of the nanostructures which is unique to each color sequence is used to identify the cells and to pair its genetic information to the already recorded morphological data.
Resultat och förväntade effekter
Parts of the project has been tested: I have collected superresolution images of the structures, demonstrated that the cell-structure attachment and structure-surface detachment chemistry works, built an aligner which can bring two samples within each others close proximity in a precisely controlled manner, up to 1 um lateral precision and was able to collect individual cells from a surface. What remains is to test the method with structures folded from a library of scaffolds with different color sequences and with known identifier sequences. This would complete a high impact work.
Upplägg och genomförande
I have proposed a chemistry for cell-structure attachment and - after some modification - I have demonstrated that it is working by confocal microscopy images. To connect the localization data of the cells and the structures, I have attempted to image the two entities together and concluded that the signal-to noise ratio with these structures is insufficient. So I have built a precision aligner to bring the previously separately imaged cells and structures in each other´s close proximity and then to collect the cells one by one for sequencing. This method also allows for live cell studies.