Optical clearing agent for bone tissue
A light-transparent agent and bone tissue technology, applied in the field of biomedical optical imaging, can solve the problems of hard bone tissue transparency, restricting the development and application of cortical optical imaging for transcranial living imaging, and achieving improved imaging depth and reduced tissue internal scattering Effect
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Embodiment 1
[0015] The scalp of a C57 mouse (20 weeks old) was cut open to expose a skull of about 1 cm × 1 cm square. According to the ratio of each component of the solution in Example 1 in Table 1, the optical transparent agent is configured into a mixed solution, which is directly added dropwise to the skull of the living mouse, and spread evenly (0.5-0.8ml / cm 2 ). figure 1 The CCD images taken before and after the treatment of the living mouse skull with light clearing agent are given. in figure 1 (A) The skull of a normal C57 mouse is exposed. Due to the opacity of the skull, it is impossible to see the intracranial cortical blood vessels; figure 1 (B) 5 minutes after adding the optical clearing agent, the skull becomes transparent to light, the cortical blood vessels below the skull become clearly visible, and some small branches can also be distinguished in the field of vision.
Embodiment 2
[0017] The isolated skull was taken from a 4-week-old SD rat, which was covered on the encapsulated fluorescent beads solution, and observed with a fluorescent microscope. The optical transparent agent is configured according to the ratio of the components of the solution in Example 2 in Table 1, and the rat skull is soaked in vitro (1.5-2ml / cm 2 ) in the light clearing agent, and after 5 minutes, cover the encapsulated fluorescent beads solution for imaging. The light-clearing effect can be reversed by flushing the skull treated with light-clearing agent with saline, and then imaging under a fluorescent microscope. figure 2 The fluorescence signals observed without skull, with skull, after light transparency of skull and after reversal of light transparency with saline are given respectively. figure 2 (A) shows the fluorescence signal observed without the skull, and the fluorescence information at this time is very strong; from figure 2 (B) It can be seen that the skull ...
Embodiment 3
[0019] Cut the brain slices of GFP-labeled transgenic mice to a thickness of 100 μm. One group was not treated with optical clearing agent and observed directly under a fluorescent microscope; solution, directly applied to mouse brain slices (0.2-0.4ml / cm 2 ), observed with a fluorescence microscope after 1 min. image 3 Respectively given are before the action of the light clearing agent under the 4 times objective lens ( image 3 (A)) after ( image 3 (B) Fluorescence signal of mouse brain slices. Only a small amount of neuron cell bodies could be observed in rat brain slices that were not treated with light clearing agent; however, after short-term treatment with light clearing agent, the cell bodies of nerve cells became obviously brighter, and the dendrite structure was now clearly identifiable.
[0020] The different optical clarity agent formulations shown in Table 1 can be formulated and used according to the methods shown in Examples 1 to 3 above to achieve these e...
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