The present disclosure relates to a Mitochondria-Associated endoplasmic reticulum Membrane (MAM)-specific fluorescence calcium sensor and the use thereof. The present disclosure can surmount the limitations of a conventional technique in that verification of calcium migration through MAM requires separate measurements of calcium ion concentrations within ER and mitochondria and situational explanations of the phenomena, and can directly measure concentrations in the paths through which calcium ions move to exclude influences on calcium ion changes through numerous different calcium ion channels existing in mitochondria, thereby providing a convenient and accurate MAM-specific calcium ion sensor.