11Britton Chance Center for Biomedical Photonics & MoE Key Laboratory for Biomedical Photonics,Wuhan National Laboratory for Optoelectronics,Huazhong University of Science and Technology,Wuhan 430074,China
12Advanced Biomedical Imaging Facility,Huazhong University of Science and Technology,Wuhan 430074,China
Dongyu LI, Tingting YU, Jingtan ZHU, Dan ZHU. In vivo Skull Optical Clearing Technique and its Applications(Invited)[J]. Acta Photonica Sinica, 2022, 51(8): 0851514
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Fig. 5. Skull optical clearing window combined with photodynamic effect for blood brain barrier opening and two-photon fluorescence observing of the leakage of Rhodamine-Dextran in blood vessels [62]
Fig. 6. Skull optical clearing window for establishment of targeted photothrombosis and evaluation of thrombolytic effect of urokinase for vessels with different sizes [65]
EDTA,dimethyl sulfoxide,sorbitol,laurinol,alcohol,glucose and weak alkaline substances
Application of the optical clearing agent on the skull
25 min
Imaging of cortical blood vessels
No assessment
SOCW[53]
Solution 1:10% collagenase
Solution 2:10% EDTA disodium
Solution 3:80% glycerol
For mice aged P15-P20:the intact skull was topically treated with solution 1 for 5~10 min;Then,solution 3 was dropped on the skull.
For mice aged P21-P30:the intact skull was topically treated with solution 2 for 5~10 min;Then,solution 3 was dropped on the skull.
For mice aged more than P30:the skull was thinned down to about 100 μm and treated with solution 2 for 5~10 min. Then,solution 3 was dropped onto the skull
5~10 min
Imaging of dendritic spines of neurons at a depth of 0~80 μm
Repeated optical clearing for 2~3 times
Targeted laser ablation of neurons
Immune response of microglia or astrocytes in the cortex was not activated
USOCA[54,61-65]
Solution 1:saturated urea solution dissolved in 75% ethanol
Solution 2:SDBS prepared by mixing 0.7 M NaOH with dodecyl benzene sulfonic acid
Solution 1 was applied to the intact skull for 10 min. Then,solution 2 was applied for 5 min
15 min
Imaging of nerves and blood vessels at a depth of 0-300 μm
Repeated daily optical clearing imaging for a week
Repeated monthly optical clearing imaging for 5 months.
Realization of blood-brain barrier opening combined with photodynamic effect or establishment of targeted photothrombosis
Immune response of microglia or astrocytes in the cortex was not activated.
No effect on the structure of the liver and kidneys
SOCW+USOCA
Solution 1:saturated urea solution dissolved in 75% ethanol
Solution 2:SDBS prepared by mixing 0.7 M NaOH with dodecyl benzene sulfonic acid
Solution 3:10% EDTA disodium
Solution 4:80% glycerol
The intact skull was treated with solution 1 for 20 min,solution 2 for 5 min,solution 3 for 25 min and solution 4 for 5 min in sequence
55 min
Imaging of blood vessels in the cortex at a depth of 900 μm combined with three-photon imaging
No assessment
VNSOCA[51]
Solution 1:saturated urea solution dissolved in 75% ethanol and deuteroxide
Solution 2:SDBS prepared by mixing 0.7 M NaOH and deuteroxide with dodecyl benzene sulfonic acid
The intact skull was topically treated with solution 1 for 10 min and solution 2 for 5 min
15 min
Compatible with visible-NIR-Ⅱ band
Imaging of cortex at a depth of 650 μm combined with third harmonic generation excited by femtosecond laser in the NIR-Ⅱ band
Establishment of targeted hemorrhagic stroke model induced by NIR-Ⅱ laser
No assessment
Table 1. Comparisons of skull optical clearing techniques
Dongyu LI, Tingting YU, Jingtan ZHU, Dan ZHU. In vivo Skull Optical Clearing Technique and its Applications(Invited)[J]. Acta Photonica Sinica, 2022, 51(8): 0851514