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

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

Solution 1：saturated urea solution dissolved in 75% ethanol

Solution 2：SDBS prepared by mixing 0.7 M NaOH with dodecyl benzene sulfonic acid

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

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

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

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