Fluorescence correlation spectroscopy detection method
A spectral detection and fluorescence-related technology, applied in the field of fluorescence-related spectral detection, can solve the problem of not being able to detect the complete process of dynamic reactions, and achieve the effect of broadening the time scale
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Embodiment 1
[0049] Example 1 The fluorescence luminescence scintillation kinetics of ATTO655 fluorescent dye molecule 5 was studied by using the fluorescence correlation spectroscopy detection method.
[0050] The ATTO655 fluorescent dye molecule 5 flashes between a fluorescent and non-fluorescent state, and the presence of reducing agents and oxidizing agents such as ascorbic acid (AA) and methyl viologen (MV) accelerates this process. Take the scintillation kinetics of ATTO655 fluorescent dye molecule 5 under different concentrations of AA and MV as an example, see Figure 9, the fluorescence spectrum detection method described in this embodiment can realize the detection of the intramolecular dynamics process on the time scale from 1.1ms to 168ms.
[0051] The specific experiments are as follows:
[0052] Slide 1 suitable for microscopy imaging was treated with aminosilane reagent (1 ml aminopropyltriethoxysilane, 5 ml acetic acid, and 94 ml methanol) overnight at room temperature, th...
Embodiment 2
[0055] Example 2 The kinetics of DNA double-strand formation was studied by using the fluorescence correlation spectroscopy detection method.
[0056] In this example, the fluorescence correlation spectroscopy detection method is combined with fluorescence resonance energy transfer, the target molecule 3 single-stranded DNA molecule labeled with Alexa488 fluorescent dye molecule 5 is used as a fluorescent donor, and the complementary effect of Cy5 fluorescent dye molecule 5 labeled Molecular 3' single-stranded DNA molecule is used as a fluorescent receptor to study the kinetics of target molecule 3 single-stranded DNA and its complementary action molecule 3' to form double-stranded DNA and DNA end respiration. see Figure 10 , the fluorescence spectroscopy detection method described in this embodiment can realize the kinetic detection of the intermolecular interaction in a large time scale range spanning three orders of magnitude from 0.45ms to 3.78s.
[0057] The specific ex...
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