Method and apparatus for tracking the motion of molecules in a microscope

摘要

An apparatus and method for measuring the mobility of molecules in a sample are disclosed. Molecules in a sample are tagged with a dye having active and inactive states that are generated by exposing dye to light of an activation wavelength and an inactivation wavelength, respectively, the activation wavelength being different from the inactivation wavelength. The sample is illuminated in a microscope with a light pattern that includes a first region in which the dye is activated and a second adjacent region in which the first dye is inactivated. After the sample is so illuminated, an image of the activated first molecules is recorded when the first molecules are illuminated with light of an excitation wavelength. Molecules having the first dye in the inactive state are distinguishable from molecules having the first dye in the active state in the microscope when illuminated with light of the excitation wavelength.

主权项

1. A method for measuring the mobility of molecules comprising:
tagging first molecules in a sample with a first dye, said first dye having an active state that is generated by exposing said first dye to light of an activation wavelength and an inactive state that is generated by exposing said first dye to light of an inactivation wavelength; simultaneously illuminating said sample with light in first and second regions in a microscope with a light pattern in which said first dye is activated by light of an excitation wavelength and substantially all of said molecules tagged with said first dye in said first region are in said active state and in which said first dye is inactivated by light of an inactivation wavelength and substantially all of said molecules tagged with said first dye in said second region are in said inactive state, to generate regions of activated and inactivated first molecules; and recording an image of said activated first molecules while said sample is no longer illuminated by said light pattern and when said first molecules are illuminated with light of an excitation wavelength, wherein molecules having said first dye in said inactive state are distinguishable from molecules having said first dye in said active state in said microscope when illuminated with light of said excitation wavelength.