Single Molecule Microbiology

It’s that time of year, so the Single Molecule Microbiology lab (Zach Hensel, Sara Costa and Cleusia Manuel) got together and designed and 3d-printed holiday lights based on the x-ray structure of the GFP variant mTurquoise2.

If you want to do the same for your own lab, check out our design and design tips at our github repository. For a more practical application of 3d printing, you’ll also find new designs that summer student Joana Valério made for a linear stage for positioning our sCMOS camera at this repository.

Congrats and good luck to Teresa Carvalho and Ricardo Quiteres who submitted their Masters theses in the last two months and will be defending in the next month! Teresa studied how bacterial effectors influence actin filamentation with a combination of in vitro and in vivo approaches (pictured left) and Ricardo studied intermediate filament defects associated with a neurodegenerative disorder by superresolution microscopy (pictured right).

João Silva’s Masters research culimated in a paper published in mSphere today: Plasmids for Independently Tunable, Low-Noise Expression of Two Genes. The paper reports the development and characterization of two compatible plasmids that we co-transformed into E. coli to independently induce GFP (GFPmut2) and RFP (mScarlet-I) expression. Together with Soraia Lopes and Diogo Grilo in the lab, we also showed how the new IPTG-inducible plasmid that João created can be used for imaging single mRNA molecules.

After almost a year and a half, Soraia’s time in our lab came to an end this week. She was instrumental in getting the lab off the ground and contributed to many on-going projects in RNA detection. She’s off to work in regenerative medicine approaches to ALS treatment!

We’ve upgraded our microscopy setup to add LED illumination and allow for rapid modulation of 5 laser lines using Micro-Manager. Plans to reproduce what we’ve built are available at the github repo.

Our design builds on several other free/open components that are described in github readme.