Three-dimensional Low Temperature Imaging Of The Whole Organ

Energy metabolism observations are of great significance for revealing many physiological and pathological mechanisms. Low-temperature optical microscopy imaging system, combining low temperature environment with optical microscopy imaging system,break through the limit of optic imaging depth by machinery milling and therefore provide a method to obtain deep biological tissue energy metabolism in a high accuracy. However, now available low-temperature optical microscopy imaging system have two technical issue: firstly, low-temperature environment contribute to prevent deformation of sample, fix sample metabolic state and enhance endogenous autofluorescence brightness of sample,simultaneously to reduce the imaging quality of system and stability of image; secondly, now available low-temperature imaging system cannot efficiently achieve threedimensional structure information and three-dimensional metabolic information of biological sample because of non-automation.This article primarily spread the discuss surrounding the first issue, influence of low temperature environment on imaging quality is reflected in three aspects: firstly, low temperature will make sample section produce ice crystals; secondly, low temperature will make objective lens produce mist; thirdly, closed liquid nitrogen circulation environment will make sample section produce fragment. Influence of low temperature environment on imaging stability is reflected as follows: liquid level fluctuation as evaporation of liquid nitrogen in sample sink will convert to temperature fluctuation of sample section, and then temperature fluctuation will influent fluorescence intensity furtherly. In order to improve system imaging quality and image stability, we design pneumatic liquid nitrogen sprinkling can device, objective circulating heating device and sample sink automatic fluid infusion device for this system.This article next spread the discuss around the second issue, to realize system automation and improve the efficiency of system imaging, we design optic automatic imaging project and pneumatic liquid nitrogen sprinkling can alarm device for this system, respectively. Finally, the above devices communicate with the other module of this system achieve the whole system automation.This article finally demonstrates level of achieve three-dimensional structure information and three-dimensional metabolic information of improved system through a set of mice brain white light and a set of mice kidney fluorescence.