Semiautomatic Method And Device For Plasticembedding Of Biological Samples

Human beings have never stopped exploring the brain,but it is still unable to explain the mechanism of the brain.And neurodegenerative diseases has become a burden on human health.So we are eager to know how the brain works.As the basic unit of the brain,neurons are the basic scientific research of brain research.However,due to the size of neurons across multiple measurements with six orders of magnitude.The need of modern neuroscience is a tool that can obtain the fine structure of sub-micron level in the whole brain.Our lab had developed the fluorescence Micro-Optical Sectioning Tomography.This system can acquire long-range projection of axons and structure of dendritic spines in the whole brain.Due to the need for imaging,the sample must be plastic embedded in order to meet the needs of precision cutting and imaging stably for a long time.To this need,our lab developed the method of plastic embedding of large volume sample,and it has been widely used to the whole brain imaging of mouse.But this protocol contains thirty steps and continue for 158 h,and all of the work is complete by hand.This cause the difficulty of condition unification of sample preparation and may reduce the efficiency of scientific research.In order to simplify the operation and to enhance the quality of the sample,this paper puts forward a set of semiautomatic sample preparation plan.Through the analysis of the manual operation,we designed the automatic change and match device of the liquid and automatic temperature constant device with the knowledge of the circuit design and mechanical engineering technology.The first device has realized the automatic processing of rinse,dehydration,infiltration.The other device has realized the automatic temperature control.Researchers can embedded the sample easily with the help of these two device,in another word we have realized the semiautomatic plan.The semiautomatic plan of plastic embedding solves the problem of complex operation,which not only improves the work efficiency,but also unifies the parameters of sample preparation.The device laid the foundation for further stabilization of imaging quality.