Optical microscope device, microscopic observation method and computer program for microscopic observation using single light-emitting particle detection technique

An optical microscopic observation method of detecting light from a light-emitting particle in a sample liquid to detect the light-emitting particle, using an optical system of a confocal microscope or a multiphoton microscope, comprising the steps of:

• (a) moving a position of a light detection region continuously multiple times within each observed subregion obtained by dividing a region to be observed within a field of view of the microscope into plural regions;
• (b) detecting light from the light detection region by a light detector;
• and
• (c) generating time series light intensity data of the light from the light detection region detected by the light detector while moving the position of the light detection region in each observed subregion, detecting a signal having a profile of a light intensity variation indicating light from the light-emitting particle in the time series light intensity data, and determining a position of the light-emitting particle corresponding to the detected signal in the region to be observed, the position being at least an X coordinate and a Y coordinate of a subregion, in the region to be observed, in which the light-emitting particle is detected.

A computer readable storage device having a computer program product including programmed instructions for observation with an optical microscope for detecting light from a light-emitting particle in a sample liquid to detect the light-emitting particle, using an optical system of a confocal microscope or a multiphoton microscope, said programmed instructions causing a computer to perform the steps of:

• moving a position of a light detection region continuously multiple times within each observed subregion obtained by dividing a region to be observed within a field of view of the microscope into plural regions;
• detecting light from the light detection region by a light detector;
• and
• generating time series light intensity data of the light from the light detection region detected by the light detector while moving the position of the light detection region in each observed subregion, detecting a signal having a profile of a light intensity variation indicating light from the light-emitting particle in the time series light intensity data, and determining a position of the light-emitting particle corresponding to the detected signal in the region to be observed, the position being at least an X coordinate and a Y coordinate of a subregion, in the region to be observed, in which the light-emitting particle is detected.