SU122305A1 - Fluorescence Microscope for Lifetime Study of Animal Organs - Google Patents

Description

Fluorescence microscopes are used for the in vivo study of animal organs.

The proposed microscope is more sophisticated in comparison with the known microscopes of this kind, since it allows one to increase the contrast of the image of an object by eliminating the scattered light of the fluorescence.

This is achieved due to the fact that a combined light filter is installed in the exit pupil of the microscope objective, dividing the aperture, on the one hand, on the outer ring, in the zone of which the object is illuminated, and, on the other hand, on the central one, which is used for observations. .

FIG. 1 and 2 shows the scheme of the proposed microscope.

FIG. 1 the illumination of the object / rays that excite its fluorescence is produced through the lens of the microscope with the help of a special optical illuminator 2 with an interference dividing mirror. The microscope lens has a thickened front lens 3, abutting the outer surface of the animal under study, moistened with physiological saline (for carrying out the optical interface between the lens and the organ).

The thickness of the front lens 3 is designed so that the focus of the microscope on the object side is inside the studied organ at a small distance from its surface facing the microscope. Accurate focusing of a certain layer of cells in the field of view of a microscope is achieved by moving the focus inside the object by moving the ocular along its optical axis or by moving lens 4 in the microscope tube.

The front lens 3 can be given the shape of a truncated cone for better penetration of the lens into the body under study.

JVb 122305

The powerful scattered fluorescence light is eliminated by a two-color light filter placed in the exit pupil of the lens, dividing the aperture into two parts: an annular, in the zone of which the objective is illuminated, and a central one, which is used for observation.

The outer ring of the light filter 5 transmits only the blue rays, and the central circle - all the rays of the visible, except for the blue (yellow filter). In addition, in the illumination system of the microscope, as usual, a blue filter b is placed, and in the ocular part there is a protective yellow filter 7 crossed with it.

Under these conditions, illumination of the object} with light that excites the fluorescence of the drug (in this case blue) is produced by the hollow cone of the beam 8 (Fig. 2) within the outer part of the objective aperture corresponding to the blue ring 9, and the observation within the inner part corresponding to , yellow circle 10. The observer examines a thin layer of cells against the background of the upper and lower dark cones 11 and 12, whose Q limits do not excite the object fluorescence; the fluorescence light emitted outside these cones due to the blue ring filter does not enter the eye of the observer or on the photographic plate. Since it is not necessary to sharply divide the aperture of the lens into two parts, it is possible to place the color filter not exactly in the pupil of the lens, but to pull it out of the lens.

A similar effect can also be obtained by placing a central blend of the corresponding diameter in the plane of the aperture stop of the illuminator and the aperture limiting the size of the pupil in the exit pupil of the microscope above the oculum.

Subject invention

A fluorescent microscope for the intravital radiation of animal organs, characterized in that, in order to increase the image contrast of the object by eliminating the scattered fluorescence light, a combined light filter is installed in the exit pupil of the microscope objective, dividing the aperture, on the one hand, on the outer annular, , the object is illuminated, and, on the other hand, on the central one, which is used for observations.