Radiometric calibration of a multiphoton microscope capable of measuring absolute photon flux of single photon...

Precise photon flux measurement of single photon sources (SPSs) is essential to the successful application of SPSs. In this work, a novel method, to our knowledge, was proposed for direct measurement of the absolute photon flux of single photon sources with a femtosecond laser multiphoton microscope. A secondary 2-mm-diameter aperture was installed under the microscope objective to define the numerical aperture (NA) of the microscope. The defined NA was precisely measured to be 0.447. An LED-based miniaturized integrating sphere light source (LED-ISLS) was used as a standard radiance source to calibrate the photon flux responsivity of the multiphoton microscope, with the defined NA. The combined standard uncertainty of the measured photon flux responsivity was 1.97%. Absolute photon flux from a quantum-dot based emitter was measured by the multiphoton microscope. The uncertainty of the photon flux was evaluated to be 2.1%. This work offers a new, to our knowledge, radiometric method for fast calibration of photon flux responsivity of microscopes, and absolute photon flux calibration of single photon sources.