A method for observing and determining the size of individual molecules and for
determining the weight distribution of a sample containing molecules of varying
size, which involves placing a deformable or nondeformable molecule in a medium,
subjecting the molecule to an external force, thereby causing conformational and/or
positional changes, and then measuring these changes. Preferred ways to measure
conformational and positional changes include: (1) determining the rate at which
a deformable molecule returns to a relaxed state after termination of the external
force, (2) determining the rate at which a molecule becomes oriented in a new direction
when the direction of the perturbing force is changed, (3) determining the rate
at which a molecule rotates, (4) measuring the length of a molecule, particularly
when it is at least partially stretched, or (5) measuring at least one diameter
of a spherical or ellipsoidal molecule. Measurements of relaxation, reorientation,
and rotation rates, as well as length and diameter can be made using a light microscope
connected to an image processor. Molecule relaxation, reorientation and rotation
also can be determined using a microscope combined with a spectroscopic device.
The invention is particularly useful for measuring polymer molecules, such as nucleic
acids, and can be used to determine the size and map location of restriction digests.
Breakage of large polymer molecules mounted on a microscope slide is prevented
by condensing the molecules before mounting and unfolding the molecules after they
have been placed in a matrix.