Protecting method for fluorescence signal of fluorescent protein in tissue sample and dehydration composition
A composition and sample technology, which is applied in the fields of biomedicine and transparent specimen preparation, can solve the problems of unfavorable sample stability observation and long-term research, fluorescent protein signal quenching, etc., and achieve the effect of good transparency and difficult quenching
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Embodiment 1
[0043] Prepare 50% tetrahydrofuran, 70% tetrahydrofuran, 80% tetrahydrofuran, 90% tetrahydrofuran, and 97% tetrahydrofuran with distilled water, each 50ml, put them in sealed glass bottles, and wrap the glass bottles tightly with aluminum foil to avoid light. Soak the brain specimens in the THF dehydrating agent from low concentration to high concentration for 12 hours each, and then replace the fresh 97% THF solution twice, and then use benzyl ether as a transparent agent to transparentize the brain specimens. Take it out from the 97% tetrahydrofuran solution and immerse it in a sealed glass bottle filled with 50ml of benzyl ether solution for 2 days, and the glass bottle is also wrapped with aluminum foil to avoid light. figure 1 and figure 2 They are respectively the transparency effect diagram of the fluorescent protein transgenic mouse brain specimen treated by Example 1 of the present invention and the light sheet illumination microscope imaging image 1 day after the tr...
Embodiment 2
[0045] Prepare 50ml of 50% ethanol, 70% ethanol, 80% ethanol, 90% ethanol, and 98% ethanol with distilled water, put them in sealed glass bottles, and wrap the glass bottles tightly with aluminum foil to avoid light. The brain specimens were soaked in ethanol dehydrating agents from low concentration to high concentration for 12 hours each, and then replaced with fresh 98% ethanol solution twice. Then use benzyl ether as a transparent agent to transparentize the brain specimen. Take the brain specimen out of the 97% ethanol solution and immerse it in a sealed glass bottle containing 50ml of benzyl ether solution for 1 day. The glass bottle is also treated with aluminum foil. Wrap away from light. image 3 It is the light-sheet illumination microscope imaging image of the fluorescent protein transgenic mouse brain specimen 3 months after being treated in Example 2 of the present invention.
[0046] The fluorescence signal of fluorescent protein remains intact after being trans...
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