6种光学透明化方法在大鼠脑组织块的透明效果比较

Comparison of transparent effects of six different optical clearing methods on rat brain tissues

目的比较6种光学透明化方法对大鼠脑组织块的透明效果。方法选取14只SD大鼠脑分别采用iDISCO、SeeDB、CUBIC、SCALEVIEW-A2、CLARITY-CUBIC、Passive-CLARITY的方法进行透明化处理。结果对照组PBS透明灰度值为13.031±0.586,iDISCO、SeeDB、CUBIC、SCALEVIEW-A2、CLARITY-CUBIC、passiveCLARITY分别为6.447±0.574、11.690±0.909、2.318±0.986、8.118±1.026、8.591±0.384、4.198±0.182,除SeeDB组外(P=0.185),其余各组均较PBS组差异均有显著性(P<0.01),CUBIC分别与各组有显著性差异(P<0.01)。透明化处理后,iDISCO、SeeDB、CUBIC、SCALEVIEW-A2、CLARITY-CUBIC、Passive-CLARITY面积改变分别为(-30.02±2.39)%、(19.74±4.09)%、(14.7±3.92)%、(10.7±5.55)%、(23.01±4.19)%、(66.51±5.68)%,各组面积改变均较iDISCO组、Passive-CLARITY组有显著性差异(P<0.01)。结论除SeeDB外,各组方法均能达到透明效果,CUBIC方法在大鼠脑组织块的透明效果优于其他各组,iDISCO处理后面积缩小,而Passive-CLARITY处理后变得膨胀。

Objective To compare the transparency efficiency of six different optical clearing method on the rat brain tissues.Methods Brain tissue blocks of 14 SD rats were processed with iDISCO,SeeDB,CUBIC,SCALEVIEW-A2,CLARITY-CUBIC,Passive-CLARITY clearing method,respectively.Results The gray value of PBS group was 13.031±0.586,that of iDISCO,SeeDB,CUBIC,SCALEVIEW-A2,CLARITY-CUBIC,passive-CLARITY clearing were 6.447±0.574,11.690±0.909,2.318±0.986,8.118±1.026,8.591±0.384,4.198±0.182,respectively.Except the SeeDB group(P=0.185),the rest groups showed significant differences compared with the PBS group(P<0.01),and there were significant differences between CUBIC and other groups(P<0.01).After the clearing treatment,the changes of tissue area ratio in the iDISCO,SeeDB,CUBIC,SCALEVIEW-A2,CLARITY-CUBIC,Passive-CLARITY method were(-30.02±2.39)%,(19.74±4.09)%,(14.7±3.92)%,(10.7±5.55)%,(23.01±4.19)%,(66.51±5.68)%,respectively.Each group showed a significant difference compared with the groups iDISCO and the Passive-CLARITY,P<0.01.Conclusions Except the SeeDB method,all the clearing methods can achieve a transparent effect,while CUBIC is better than the other groups applied for rat brain tissues.The tissue block volume is shrunken after iDISCO clearing,and expanded after Passive-CLARITY processing.