Same cells, same fixative, same immunostaining protocol for microtubules but one little thing is making all the difference in the amount of non-specific stain localized using dSTORM. Details to come, hopefully in our soon-to-be-submitted protocols and artefacts paper.
We are back from Germany! After a quick detour to visit the Sauer lab in Wurzburg, Toby and Donna met up with Rosey and Heather in Berlin for the PicoQuant Single Molecule Workshop. Posters were presented with lots of good feedback and Toby gave the shortest talk of his life. It was fantastic to see so many different labs from all over the world presenting state-of-the-art fluorescence research and in between the onslaught of information that the conference provided many ideas for new and ongoing research projects were hatched. A river cruise in beautiful weather and dinner at the top of the Radio tower capped off a truly wonderful trip.
The first cell line we ever tried to image is back and yielding some stunning whole-cell 2D images of the actin network around the nucleus. The glutaraldehyde protocol we have specifically optimized for actin ultrastructure preservation has now been tested on three different mammalian cell lines, all with great results, and should be published as part of an upcoming paper.
Almost time for the 20th PicoQuant Single Molecule Workshop. Four posters will be present spanning nanocrystals to Rabies, pH sensors to imaging artefacts. Heather will be arriving in Berlin via Denmark along with Bell group alumni Emma Hooley who is now doing a postdoc at the University of Coperhagen. Rosey will be flying in direct from Melbourne. And Donna and Toby will be coming up from Wurzburg along with members of Markus Sauer’s group.
Certainly a good excuse to touch base with our dSTORM collaborators and revisit Wurzburg once more…
This week we had the pleasure of hosting several science-savvy Year 10 work-experience students and were tasked with showing them how a working research lab operates. In between looking at the fluorescence of fingerprints and various dust samples from around the department, we also spent time checking and calibrating the pH of the stock solutions and imaging buffers in use with our favorite high-tech pH paper. Pretty cool work experience if you ask me.
Stretched DNA labelled using YOYO-1 and imaged on the home built super resolution setup with varying success. Ongoing work by Donna and Lukas has confirmed that sub-diffraction imaging of DNA is really, really, really hard but absolutely possible. Some great images have been captured. Sometimes, as in the middle, the DNA isn’t so well stretched or labelled. But other times, as in the top and bottom images, a magnitude-improved spatial resolution is demonstrated in the double strand cross-sections.
20th International Workshop on
“Single Molecule Spectroscopy and Ultra Sensitive Analysis in the Life Sciences”
Abstracts have been submitted and registrations paid. Toby, Donna, Rosey and Heather are all excited to be heading to Germany for this fantastic meeting!