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Article summary:
1. To observe complex biological systems, high-throughput imaging methods are needed to capture fast biological events.
2. Multiphoton microscopy is the gold standard for deep tissue recording in live mammalian models, but it is often too slow to keep up with these fast dynamics.
3. An optical gearbox has been developed to adapt commonly employed laser sources and scanners for high frame rate imaging, overcoming the scan frequency and scan throughput tradeoff.
Article analysis:
The article provides a comprehensive overview of the current state of multiphoton functional imaging and its limitations in terms of speed and depth. The authors provide a detailed description of the various factors that affect the speed of multiphoton imaging, such as excitation power, pulse repetition rate, scanner speed, excitation numerical aperture (NA), fluorescence signal collection efficiency, fluorophore concentration within the focal volume, fluorophore brightness and bleaching rate, photodamage threshold, and thermal limit. They also discuss several new techniques that have been developed to achieve high frame rate recording.
The article is generally reliable and trustworthy in its presentation of information; however there are some potential biases that should be noted. For example, while the authors do mention some existing techniques for achieving high frame rate recording (such as parallel recording via cameras), they focus primarily on their own proposed solution – an optical gearbox – which could be seen as a form of self-promotion or bias towards their own work. Additionally, while they do mention some potential drawbacks to their proposed solution (such as reduced nonlinear excitation efficiency), they do not explore any other possible counterarguments or risks associated with it in detail. Furthermore, while they provide evidence for their claims regarding multiphoton imaging speeds (such as citing relevant studies), they do not provide any evidence for their claims regarding the efficacy or advantages of their proposed solution over existing techniques.
In conclusion, this article provides a comprehensive overview of multiphoton functional imaging and its limitations in terms of speed and depth; however there are some potential biases that should be noted when considering its trustworthiness and reliability.