Laser scanning microscope system
View Patent ↗An immersion microscope objective formed of thirteen or fewer lens elements includes, in order from the object side, first and second lens groups of positive refractive power, a third lens group, a fourth lens group having negative refractive power with its image-side surface being concave, and a fifth lens group having positive refractive power with its object-side surface being concave. The first lens group includes, in order from the object side, a lens component that consists of a lens element of positive refractive power (when computed as being in air) and a meniscus lens element having its concave surface on the object side. Various conditions are satisfied to ensure that images of fluorescence, obtained when the immersion microscope objective is used in a laser scanning microscope that employs multiphoton excitation to observe a specimen, are bright and of high resolution. Various laser scanning microscopes are also disclosed.
1. A laser scanning microscope system comprising:
a laser beam deflection means for deflecting a laser beam over a scanning range onto a specimen; and
an immersion microscope objective having a pupil diameter;
wherein
an object-side numerical aperture NA of said immersion microscope objective lies within the following range
0.85<NA<1.15, and
an effective diameter of the laser beam which is set by the laser beam deflection means is substantially equal to a pupil diameter at the pupil of said immersion microscope objective.
2. The laser scanning microscope system according to claim 1 , and further comprising:
a pupil projection lens; and
an imaging lens;
wherein the following condition is satisfied
0.9<(2×NA× f )/( S ×ft1/fp1)<1.1
where
NA is the object-side numerical aperture of said immersion microscope objective,
f is the focal length, in mm, of said immersion microscope objective,
S is the size, in mm, of the effective diameter of said laser beam deflection means,
ft1 is the focal length, in mm, of said imaging lens, and
fp1 is the focal length, in mm, of said pupil projection lens.
3. The laser scanning microscope system according to claim 2 , and further comprising:
a non-confocal detection optical system;
wherein
the non-confocal detection optical system has a detection field of view that is larger than the scanning range of said laser beam deflection means.
4. The laser scanning microscope system according to claim 3 , wherein:
NAe>0.06,
where
NAe is a numerical aperture of said non-confocal detection optical system in ray tracing from the pupil.
5. The laser scanning microscope system according to claim 3 , and further comprising:
a non-scanning illumination introducing optical system; and
said non-confocal detection optical system and said non-scanning illumination introducing optical system are provided independently.
6. The laser scanning microscope system according to claim 3 , wherein said non-confocal detection optical system includes:
a first pupil projection lens that converts a light beam from the pupil of said immersion microscope objective to a nearly collimated, but somewhat converging, light flux;
a second pupil projection lens that projects said somewhat converging light flux onto a detector; and
a laser cut-off filter and a dichroic mirror that are positioned between said first pupil projection lens and said second pupil projection lens.
7. The laser scanning microscope system according to claim 1 , wherein at least a lens group in said immersion microscope objective is movable along an optical axis.
8. The laser scanning microscope system according to claim 1 , wherein:
a scanning optical system, along an optical path from said laser beam deflection means to said immersion microscope objective, and said non-confocal detection optical system are provided independently; and
a means for splitting the optical path between said scanning optical system and said non-confocal detection optical system is provided on the image side of said immersion microscope objective.