Dual-color Imaging Using Acousto-optic Deflector With Dynamic Compensation Method

Optical imaging methods for biological research has been paid attention to and loved by biologists because of its low cost and high resolution. Automated optical imaging methods can get fine three-dimensional structures with high accuracy which liberates the hands of biologists for making the imaging of biological samples of large volume no more indispensable of a lot human resources to frequently slicing, imaging and registration. Confocal fluorescence microscopy optical sectioning tomography system(confocal-f MOST) devotes to acquire the fluorescent labeled mouse brain in three-dimensional with high accuracy, can get the nerves of long-range projection in whole brain and provides a powerful tool for neurobiologists to study brain structures and functions. The development and maturity of multicolor labelling and imaging technology makes biological studies more scientific and effective, but current confocal fluorescence microscopy optical sectioning tomography system only used for data acquisition of a single color labeled sample. Due to the deflector based on diffraction effect and the scanning polychromatic beams can't coincide, the implemented of multi-color confocal-f MOST has certain technology difficulties.In this paper, multicolor confocal fluorescence micro-optical sectioning tomography system has been focused, based on research of single color imaging system. The deflection angle inconsistencies which exists when acousto-optic deflector scans multicolor simultaneously has been resolved, synchronous scanning method of multicolor laser beams based on dynamic compensation for acousto-optic deflector has been developed and applied to system implementation, making confocal fluorescence microscopy optical sectioning tomography system able to image biological samples labelled by two colors with dual channels simultaneously.On this basis, the discuss proposes the ideas of spinning disk multicolor confocal optical sectioning fluorescence microscopy tomography system. The possible technical solution of the proposed multi-color automation imaging system has been demonstrated, and the use of biological samples confirmed its feasibility. Through preliminary experiments the authors found that the spinning disk confocal microscopy could imaging the axons, and this paper argues that a multi-color automated system for three-dimensional fluorescence imaging set up by spinning disk shows good prospects, especially for submicron level neurite tracking.