HO1N1 cells stained with Bacmam 2.0 Mito-GFP (green), ER-tracker (blue) and SiR-DNA (red). One 3-color image was taken every 10 seconds for 1.5 hours, giving a total of 1000 images. Because the RCM is very sensitive laser power can be kept very low (few microwatts), enabling long-term imaging. The native lateral resolution of the RCM is 170 nm. After deconvolution by SVI Huygens the resolution is improved to 120 nm!
Movie taken by Jeroen Kole (Confocal.nl), sample courtesy Dandan Ma (ACTA, Amsterdam), equipment provided by Marko Popovic (Nikon Center of Excellence, Amsterdam University Medical centers, VUmc, Amsterdam.)
HO1N1 cells expressing Mitochondria-RFP through the Bacmam expression system. One image was taken every 10 seconds for 61 hours (giving a total of almost 22,000 images!). Laser power was measured to be 1 microwatt at the sample plane. Movie taken by Jeroen Kole (Confocal.nl), sample courtesy Dandan Ma (ACTA, Amsterdam), equipment provided by Marko Popovic (Nikon Center of Excellence, Amsterdam University Medical centers, VUmc, Amsterdam.)
The first video shows the RAW data from RCM, the entire 61 hour time lapse.
The second video shows the benefits of deconvolution. We used SVI Huygens CMLE deconvolution to improve the resolution to 120 nm and have an even better signal-to-noise ratio.
Protip: watch in 1440 px HD or 4K!
HeLa cells expressing EB1 tagged with YFP. 2 seconds time interval, (original time lapse 40 minutes), acquired with 8µW laser power. Image taken by and courtesy of Andreas Kurz, Sauer Lab, University of Würzburg, Germany.
HEK 293 cells expressing two membrane-proteins. Image is a maximum-projection of a z-stack in time. Sample courtesy: Reuben Levy-Myers, Sockanathan lab, Johns Hopkins University, Baltimore, USA.
Maximum-projection of a Z-stack in time. Arabidopsis hypocotyl cells expressing GFP-EB1b – the protein binds to +ends of microtubuli and indicates their growth (so called comet-like movement). EB1b also binds to nuclear membrane, as seen by the high intensity structures. Sample courtesy: Dr S L Shaw. Acquired during the AQLM course at the MBL in Woods Hole, MA, USA.