Fluorescence control method of biological tissue labeled with pH-insensitive fluorescent protein
A control method and biological tissue technology, applied in fluorescence/phosphorescence, material analysis through optical means, material excitation analysis, etc., can solve the problems of background fluorescence interference, uncontrollable fluorescence, low axial resolution, etc., and achieve convenient activation , Fluorescence intensity contrast increases, good effect
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Embodiment 1
[0056] (1) The mouse brain labeled with the red fluorescent protein DsRed was fixed with 4% PFA solution and rinsed with 0.9% NaCl solution;
[0057] (2) The fluorescent protein in the mouse brain labeled with the fixed and rinsed red fluorescent protein DsRed was used 100mM CuSO 4 The solution (pH 4~5) is quenched for 30 minutes and fluorescence imaging is performed;
[0058] (3) Use 200 mM EDTA-Na for the fluorescent protein in the mouse brain labeled with the quenched red fluorescent protein DsRed 4 The aqueous solution (pH 11-11.5) was reactivated for 30 minutes while fluorescence imaging was performed.
[0059] Fig. 5 is a quenching-activation imaging diagram of mouse brain tissue labeled with red fluorescent protein DsRed in this example. Figure 5(a) is the red fluorescent protein DsRed labeled mouse brain slice in 100mM CuSO 4 After quenching in the solution for 30 minutes, the contrast of the image was increased 20 times in order to compare with the fluorescence intensity aft...
Embodiment 2
[0062] (1) The mouse brain labeled with the red fluorescent protein DsRed was fixed with 4% PFA solution and rinsed with 0.9% NaCl solution;
[0063] (2) The fluorescent protein in the mouse brain labeled with the fixed and rinsed red fluorescent protein DsRed was used 200mM FeCl 3 The solution (pH 3.5-4.5) was quenched for 30 minutes and fluorescence imaging was performed;
[0064] (3) The fluorescent protein in the mouse brain labeled with the quenched red fluorescent protein DsRed was reactivated with a 400mM sodium protoporphyrin aqueous solution and the pH was adjusted to 11-12 with sodium hydroxide and then reactivated for 30 minutes while performing fluorescence imaging .
[0065] 6 is a quenching-activation imaging diagram of the mouse brain tissue labeled with red fluorescent protein DsRed in Example 2. FIG. Figure 6(a) is the red fluorescent protein DsRed labeled mouse brain slice in 200mM FeCl 3 The image after quenching in the solution for 30 minutes, its contrast has be...
Embodiment 3
[0068] (1) The mouse brain labeled with the red fluorescent protein DsRed was fixed with 4% PFA solution and rinsed with 0.9% NaCl solution;
[0069] (2) The fluorescent protein in the mouse brain labeled with the fixed and rinsed red fluorescent protein DsRed was used 10mM CoSO 4 Adjust the pH of the solution to 3-4 with the solution and acetic acid, perform quenching treatment for 30 minutes, and perform fluorescence imaging;
[0070] (3) Use 100 mM EDTA-Na for the fluorescent protein in the mouse brain labeled with the quenched red fluorescent protein DsRed 3 The aqueous solution (pH 11-11.4) was reactivated for 30 minutes while fluorescence imaging was performed.
[0071] The fluorescence intensity of the mouse brain tissue after reactivation is equivalent to the fluorescence intensity of the mouse brain image after quenching increased by 10 times. It shows that the fluorescence quenching-reactivation control method of this embodiment can be used to reduce the fluorescence of mous...
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