This technology introduces a fifth-order multiphoton microscopy system. This advanced method enables high-resolution, label-free imaging of biological samples using nonlinear optical effects. Laser scanning multiphoton microscopy has gained prevalence across a wide range of applications due to its high-resolution imaging potential. This system uses both fifth harmonic generation and five-photon excitation signals, enabling the use of longer pump wavelengths. These longer wavelengths can be useful in a wide range of scientific applications, such as biological research, materials characterization, geologic studies, and semiconductor manufacturing. Furthermore, the system offers improved lateral and axial resolution over lower order multiphoton imaging methods. By leveraging the fifth harmonic of a laser and five-photon excitation, this technique can penetrate deeper while providing high-contrast images without the need for external fluorescent labels. Background: Traditional multiphoton microscopy methods, such as two-photon or three-photon excitation, are widely used for deep-tissue imaging due to their ability to minimize photodamage and achieve high-resolution images. However, these methods often require external fluorescent dyes, which can cause photobleaching and may alter the biological properties of samples. Second-order and third-order multiphoton imaging systems are in active use, and fourth-order multiphoton imaging has been demonstrated as well. This system further expands the range of multiphoton microscopy systems by introducing a fifth-order system. The system is capable of increased resolution over lower-order methods. Applications:
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