Photonic Science
Professional Imaging

Confocal Fluorescence Imaging.

 

Ultra low noise, cooled second generation sCMOS cameras are operated with open software and do not require a frame grabber. They come as a straightforward and affordable upgrade to existing cooled CCD camera systems struggling to deliver high dynamic range, high resolution and high frame rate all at the same time.

In a confocal microscope, the detector aperture obstructs the light that is not coming from the focal point. The out-of-focus light is blocked by the pinhole and in order to produce a crisp image with haze contribution introduced by the depth of field from the objective. The smaller the pinhole, the sharper the image will be as it will effectively block the fluorescence from the nearest neighbouring planes, but also less light will be caught by the detector. Therefore a very sensitive cameras is required to capture the fluorescence arising from the confocal plane. Typical integration time must be kept as short as possible in order to avoid cell damage under extended periods of digital recording. Typically 100ms to 1 second is used per image, depending on sensitivity settings chosen on the camera. Three dimensional re construction is achieved by acquiring multiple images at different Z positions using a piezo / stepper motor driven z axis on a motorized microscope. Z stacks acquired over time allow building multidimensional data sets taking into account multiples variables such as wavelengths, x,y,z positions and time. Multiple cameras can be synchronized as one single detector in order to produce parallel acquisition.

Confocal Fluorescence image

Confocal Fluorescence image