Dr. Philippe Bastin, Principle Investigator
Bastin Lab, Institut Pasteur, Paris
The lab of Dr. Philippe Bastin at Institut Pasteur is primarily interested in improving understanding of the trypanosome parasites, which are significant in human health due to their role in sleeping sickness. The trypanosomes also offer a useful experimental model to increase understanding of cilia and flagella function. The lab therefore does a significant amount of cell biology investigations including live cell imaging studies, four color immunofluorescence (IF) and dSTORM imaging.
Dr. Bastin’s lab studies protein trafficking in the flagella of trypanosomes. The challenge is that these proteins move quickly, at a rate of 2-5 microns per second in a very narrow (300 nm) environment, which is very close to the resolution limit for light microscopy.
Imaging is a tradeoff between time resolution and sensitivity, even at 100 ms exposure the particle has moved by half a micron during the exposure time leading to smearing of the trains, so it is quite challenging. When the train moves to the end of the flagella they then split into 3 smaller trains making them more difficult to detect.
Dr. Bastin was looking for a camera that had good sensitivity to decrease the exposure time. If the camera was too slow, it wouldn’t capture the movement. The previous EMCCD was used for its sensitivity, but this was limiting the resolution due to the large pixels. Four-color immunofluorescence imaging was also challenging, as when imaging dyes with longer wavelengths, such as Cy5, quantum efficiency is low.
The combination of increased sensitivity and signal to noise ratio meant the 95B was the best camera choice.