Microscope and method for observing biological specimen in living state

A method for observing a biological specimen in a living state, wherein the method comprises:

• adding a luminescence substrate to the biological specimen, wherein:
• the luminescence substrate includes a self-luminous fused protein that contains a luminescence enzyme and a fluorescent protein, and
• the self-luminous fused protein generates light by chemiluminescence;
• illuminating the biological specimen with a radiation pattern of a control light that changes a state of the chemiluminescence, wherein the radiation pattern has a defined shape;
• and
• detecting a light emitted from the biological specimen by chemiluminescence, wherein:
• when illuminating an observation surface of the biological specimen with the control light, the shape of the control light is defined as a first pattern, and
• when detecting the light emitted from the biological specimen by chemiluminescence, a second pattern formed on the observation surface by a mutual effect between a frequency component of the biological specimen and a frequency component of the first pattern is detected, and the frequency component of the biological specimen is determined based on the frequency component of the first pattern and a frequency component of the second pattern.