One-scan fluorescence emission difference nanoscopy developed with excitation orthogonalized upconversion...

Area of Science:

• Nanotechnology
• Optical Microscopy
• Materials Science

Background:

• Super-resolution microscopy aims to overcome the diffraction limit of light.
• Fluorescence Emission Difference (FED) nanoscopy offers potential for high-resolution imaging.
• Developing faster and more versatile FED techniques is crucial for advanced imaging applications.

Purpose of the Study:

• To experimentally realize a one-scan Fluorescence Emission Difference (FED) nanoscopy technique.
• To achieve high-resolution imaging of upconversion nanoparticles using orthogonal excitation.
• To enhance the imaging speed and applicability of FED nanoscopy.

Main Methods:

• Utilized NaYF4:Er3+@NaYF4@NaYF4:Yb3+/Tm3+ core-shell upconversion nanoparticles.
• Employed synchronized laser beams: a solid 940 nm and a hollow 808 nm beam.
• Implemented orthogonal generation and collection of green (Er3+) and blue (Tm3+) emissions.

Main Results:

• Successfully achieved simultaneous imaging of two distinct color emissions.
• Obtained orthogonal excitation and collection of Er3+ green and Tm3+ blue emissions.
• Generated a super-resolution image with a resolution of 54 nm via simple subtraction.

Conclusions:

• Demonstrated a novel one-scan FED nanoscopy strategy.
• The excitation orthogonality significantly improves imaging speed and applicability.
• This method provides a powerful tool for high-resolution nanoscale imaging.