Non-contact total emission detection method and system for multi-photon microscopy

A multi-photon microscope comprising:

• an illumination source for transmitting an illumination light into a housing;
• an objective lens arrangement defining an aperture and disposed inside the housing, the objective lens arrangement being oriented in an optical pathway of the illumination source to direct the illumination light through the aperture of the objective lens arrangement to a focused illumination region of a sample under observation disposed outside the housing;
• a light collection system disposed inside housing, the light collection system having a reflector defining an aperture and arranged proximate the objective lens arrangement such that the aperture of the objective lens arrangement is oriented towards the aperture of the reflector;
• and
• a detection system oriented in an optical pathway of the objective lens arrangement and the light collection system for detection of light emitted by the sample,
• wherein the objective lens arrangement is configured to direct a first portion of emission light that is emitted from the illumination region of the sample under observation in response to being illuminated by the illumination light to the detection system;
• wherein the light collection system is configured to direct a second portion of emission light that is emitted from the illumination region of the sample under observation in response to the illumination light to the detection system.