A Super-resolution Imaging Method Based on Fluorescence Resonance Energy Transfer
A fluorescence resonance energy, super-resolution imaging technology, applied in the field of super-resolution imaging, can solve problems such as high cost and complex imaging software
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Embodiment 1
[0041] Example 1 Super-resolution imaging of Cy3-Cy5 probe-labeled Hela cell microtubules
[0042] Construction of Cy3-Cy5 fluorescent probes with high FRET efficiency: Fluorescent nanospheres were prepared by inverse microemulsion method.
[0043] Experimental materials: sodium silicate solution (water glass) (the water glass modulus is 3.1, Baume degree is 40), hydrochloric acid (concentration is 1mol / L), sodium lauryl sulfate SDS, toluene, n-amyl alcohol , deionized water (W), fluorescent dye Cy3-Cy5.
[0044] The specific steps of this technique include:
[0045] Osmotic reaction: take 0.1g surfactant SDS (S), 0.2g fluorescent dye (donor Cy3 and acceptor Cy5, donor acceptor molar ratio 1:2), 0.2ml toluene and 5ml water glass solution prepared in advance Put it into a beaker, and mix it uniformly by ultrasonic (50W ultrasonic power, 1 minute mixing time) to obtain a colostrum, add n-amyl alcohol dropwise until the system is suddenly transparent, and obtain microemulsion A...
Embodiment 2
[0055] Example 2 Super-resolution imaging of Hela cell microtubules labeled with Atto488-Atto540 probe
[0056] Construction of Atto488-Atto540 Fluorescent Probe with High FRET Efficiency
[0057] Experimental materials: sodium silicate solution (water glass) (the water glass modulus is 3.2, Baume degree is 40), hydrochloric acid (concentration is 1mol / L), sodium lauryl sulfate SDS, toluene, n-amyl alcohol , deionized water (W), fluorescent dyes Atto488-Atto540.
[0058] Take 0.1g of surfactant SDS (S), 0.2g of fluorescent dye (donor Atto488 and acceptor Atto540, donor-acceptor molar ratio 1:2), 0.2ml of toluene and 5ml of water glass solution prepared in advance into a beaker In the process, ultrasonically mix evenly (ultrasonic power 50W, mixing time 1 minute) to obtain a primary emulsion, add n-amyl alcohol dropwise until the system is suddenly transparent, and obtain microemulsion A.
[0059] Polymerization reaction: replace the water glass solution with 1mol / L HCl solut...
Embodiment 3
[0067] Example 3 Super-resolution imaging of Hela cell microtubules labeled with Atto550-Atto647N probe
[0068] Construct Atto550-Atto647N fluorescent probe with high FRET efficiency:
[0069] Experimental materials: sodium silicate solution (water glass) (the water glass modulus is 3.3, Baume degree is 40), hydrochloric acid (concentration is 1mol / L), sodium lauryl sulfate SDS, toluene, n-pentanol , deionized water (W), fluorescent dye Atto550-Atto647N.
[0070] Osmotic reaction: take 0.1g surfactant SDS (S), 0.2g fluorescent dye (donor Atto550 and acceptor Atto647N, donor acceptor molar ratio 1:2), 0.2ml toluene and 5ml water glass solution prepared in advance Put it into a beaker, and mix it uniformly by ultrasonic (50W ultrasonic power, 1 minute mixing time) to obtain a colostrum, add n-amyl alcohol dropwise until the system is suddenly transparent, and obtain microemulsion A.
[0071] Polymerization reaction: replace the water glass solution with 1mol / L HCl solution, a...
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