Dr. Christoph Baumann, Lecturer at the University of York, Department of Biology and his group work with advanced imaging techniques to push forward our understanding of spatio-temporal dynamics in the bacterial cell envelope. Using a Photometrics camera, the group was the ﬁrst to observe that, contrary to expectations, proteins in the outer cell membrane don’t diffuse signiﬁcantly when tracked, and that new proteins are inserted predominantly at mid-cell during growth.1 This means that bacteria can very quickly turnover their outer membrane proteins to adapt to new environmental challenges during growth, and this work initiated a new investigation of inter-membrane crosstalk in the Gram negative bacterial cell envelope.2
To pursue this research area, Dr. Baumann and his colleagues use TIRF microscopy alongside laser scanning confocal FRAP microscopy. When tracking single molecules, sensitivity and speed are all-important. “Better temporal and spatial resolution means better quality data,” Dr. Baumann shares. Previously, the large pixels, slow speed, and the excess noise factor of their EMCCD camera limited both these aspects. Further, the small sensor size and the need to use a 1.6× magniﬁcation optic to match pixel size lead to a very small ﬁeld of view, and using additional lenses lost them precious light.
The Photometrics Prime 95B Scientiﬁc CMOS (sCMOS) camera was the perfect ﬁt for the team’s research, with the 11 µm pixels giving optimal resolution paired with the 100× objectives used for TIRF microscopy. The faster speed and higher signal to noise ratio of the back-illuminated CMOS also provided better temporal resolution. “This increased resolution is of great beneﬁt for all the research we do, not just this experiment,” Dr. Baumann told us. “Since the camera is USB3.0, it’s very easy to move the camera to other setups for short periods.”
The huge ﬁeld of view of the Prime 95B is not only useful for increasing throughput, but allows the team to use single-camera optical splitting technology. Dr. Baumann concluded, “We’re going to use a polarization splitter in an upcoming experiment, and the Prime 95B’s chip is large enough to still have a great ﬁeld of view with both images side by side.”