The Research Of New Tissue Optical Clearing Technique

In recent years,with the rapid development of biomedical process,life sciences and medical research transits from the cell research with heavy artificial traces to the organization,embryos even model animal research,which have more physiological meaning.In order to obtain three-dimensional optical imaging of these tissues and organs,the traditional technique is to perform tissue slice re-stratification imaging,it makes the whole process time-consuming and effortless.Moreover,this slicing process damage the integrity of the organ and the continuity of certain subcellular cell structures and easily cause cell disease physiological information partial loss,thus promotes the tissue optical clearing technology emergence and rapid development.Tissue optical clearing technology,which transforms intact opaque tissue into an optically transparent construct by reducing light scattering and then can be further combined with optical imaging technology to facilitate intact tissue organ deep optical(fluorescence/absorption)three-dimensional imaging.It has been attracted increasing interest in biomedical photonics area due to their advantages compared to conventional mechanical-sectioning-based methods.These characteristics of tissue optical clearing technology make it widely applied in the field of biological medicine drug development and is playing an increasingly important role.This paper first summarizes optical clearing technology principles,main approaches and their current research situation and achievements in biomedicine,especially in the field of brain science.The comparison of their performance parameters and the further development challenge are also presented.After the experimental study carried out,the specific research contents and results are as follows:(1)Comparative researched tissue optical clearing technology which has been published on the international journal.The results show that: the fluorescent protein signal preservation effect of u DISCO is poor,the transparency of See DB in adult big mouse tissue is very low,the sample volume expansion of Scale is large,CUBIC isn't compatible with Di I dyes,and Scale S has a lower clearing rate in adult tissue samples.At the same time,for the problem that the clearing rate of See DB is relatively slow,it is proposed that the clearing rate of See DB can be improved by adding the chemical permeation enhancer agent dimethyl sulfoxide.(2)We develop a new tissue optical clearing technology?Ubas M to solve the problem of fluorescence quenching problem,cracking cell membrane structure(which can not be compatible with Di I dye),low clearing rate and large change in tissue volume shape,which can been found in the old tissue optical clearing technology.Ubas M: The reagents with good clearing function of urea,meglumine,sucrose,1,3-dimethyl-2-imidazolidinone and Triton X-100 were screened and invented with Ubas M-1 solution: 25wt%meglumine+ 25wt%urea+20wt%1,3-dimethyl-2-imidazolidinone+0.2wt%Triton X-100,Ubas M-2 Solution: 25wt%urea+40wt%sucrose+20wt%dimethyl-2-imidazolidinone,at the same time,we put forward a set of complete method for sample clearing.(3)We discuss varous characteristics of Ubas M,including clearing rate,transparency,fluorescent protein signal preservation,ability to be compatible with Di I staining(membrane structure protection),changes in tissue volume morphology,and application in biomedical research.The results show that Ubas M has less influence on fluorescent protein signal and can be compatible with Di I dye,the clearing rate is faster,the sample transparency is higher,the volume change of tissue volume is very small,the safety is good and the operation is simple and the cost is low.Meanwhile,this technology has been used in Alzheimer's disease and breast cancer research and other biomedical research and achieved good experimental results.In conclusion,compared to other tissue optical clearing technology,Ubas M is a new tissue optical clearing technology,which has a lot of advantages,such as simple,efficient,strong applicability,wide range of applications.It has great application prospect in life science and medicine and drug development.