Photon counting imaging with an electron-bombarded CCD: towards a parallel-processing photoelectronic...

We have used an electron-bombarded CCD for optical photon counting imaging. The photon event pulse height distribution was found to be linearly dependent on the gain voltage. We propose on this basis that a gain voltage sweep during exposure in an electron-bombarded sensor would allow photon arrival time determination with sub-frame exposure time resolution. This effectively uses an electron-bombarded sensor as a parallel-processing photoelectronic time-to-amplitude converter, or a two-dimensional photon counting streak camera. Several applications that require timing of photon arrival, including Fluorescence Lifetime Imaging Microscopy, may benefit from such an approach. A simulation of a voltage sweep performed with experimental data collected with different acceleration voltages validates the principle of this approach. Moreover, photon event centroiding was performed and a hybrid 50% Gaussian/Centre of Gravity + 50% Hyperbolic cosine centroiding algorithm was found to yield the lowest fixed pattern noise. Finally, the camera was mounted on a fluorescence microscope to image F-actin filaments stained with the fluorescent dye Alexa 488 in fixed cells.