Optical: systems and elements – Compound lens system – Microscope
Reexamination Certificate
2000-12-01
2004-10-26
Robinson, Mark A. (Department: 2872)
Optical: systems and elements
Compound lens system
Microscope
C359S379000, C359S380000
Reexamination Certificate
active
06809861
ABSTRACT:
This application claims benefit of Japanese Application No. Hei11-344786 filed in Japan on Dec. 3, 1999, the contents of which are incorporated by this reference.
BACKGROUND OF THE INVENTION
The present invention relates to optical apparatus and, more particularly, to fluorescence observation optical apparatus including a fluorescence microscope and a stereoscopic microscope allowing fluorescence observation.
In recent years, fluorescence observation under a fluorescence microscope and a stereoscopic microscope has been widely used not only in micro observation but also in macro observation at low magnification. In particular, fluorescence proteins such as GFP (Green Fluorescence Protein), CFP (Cyan Fluorescence Protein) and YFP (Yellow Fluorescence Protein) have advantages in comparison to conventional fluorescent dyes. That is, such fluorescence proteins show a comparatively low toxicity to cells, suffer less fading and provide brighter fluorescence. Accordingly, the use of fluorescence proteins in the field of genetic research has also increased.
Thus, objects to be observed range from cells in micro observation to individuals such as fruit flies and mice in macro observation. Therefore, there has been proposed a fluorescence observation apparatus including not only an ordinary fluorescence microscope but also a stereoscopic microscope as an apparatus allowing fluorescence observation.
The stereoscopic microscope is a microscope characterized by having a very long working distance in comparison to ordinary microscopes and allowing three-dimensional observation.
FIG. 43
shows a conventional fluorescence observation apparatus including a stereoscopic microscope. First, the observation optical system of the stereoscopic microscope has an interchangeable objective
41
and two variable magnification optical systems
42
R and
42
L associated with right and left eyes, respectively. The observation optical system further has imaging lenses
43
R and
43
L and eyepieces
44
R and
44
L. An image of a sample
47
is magnified by the objective
41
and the variable magnification optical systems
42
R and
42
L, and the magnified image of the sample
47
is viewed through the imaging lenses
43
R and
43
L and the eyepieces
44
R and
44
L.
The objective
41
and each of the variable magnification optical systems
42
R and
42
L are arranged in the form of an afocal optical system. Similarly, the variable magnification optical systems
42
R and
42
L and the imaging lenses
43
R and
43
L are arranged in the form of afocal optical systems, respectively. Thus, the observation optical system is excellent in system flexibility.
The fluorescence illumination optical system of the stereoscopic microscope has a light source
51
, an illumination lens system
52
, an excitation filter
53
, and a dichroic mirror
54
L.
Light from the light source
51
, which is a mercury lamp, is led to the excitation filter
53
through the illumination lens system
52
. Of the light from the light source
51
, only excitation light of wavelength needed to excite the sample
47
is selectively transmitted by the excitation filter
53
. Excitation light emanating from the excitation filter
53
is reflected toward the variable magnification optical system
42
L by the dichroic mirror
54
L and applied to the sample
47
through the variable magnification optical system
42
L and the objective
41
.
At the sample
47
, fluorescent light is produced from portions of the sample
47
stained with a fluorescent dye by illumination with the excitation light. The fluorescent light from the sample
47
is collected by the objective
41
and led to a right observation optical path R for an observer's right eye and also to a left observation optical path L for an observer's left eye. Fluorescent light led to the left observation optical path L passes through the variable magnification optical system
42
L and the dichroic mirror
54
L and reaches an absorption filter
55
L. The absorption filter
55
L transmits only fluorescent light of specific wavelength selected according to the spectral characteristics thereof. The fluorescent light of specific wavelength is imaged through the imaging lens
43
L and viewed as a fluorescence image through the eyepiece
44
L. Fluorescent light led to the right observation optical path R passes through the variable magnification optical system
42
R and a dichroic mirror
54
R and reaches an absorption filter
55
R. Fluorescent light passing through the absorption filter
55
R, as in the case of fluorescent light passing through the absorption filter
55
L, is imaged through the imaging lens
43
R and viewed as a fluorescence image through the eyepiece
44
R.
The arrangement of an ordinary fluorescence microscope is shown in FIG.
44
. The fluorescence illumination optical system of the ordinary fluorescence microscope has a light source
51
, an illumination lens system
52
, an excitation filter
53
, a dichroic mirror
54
, and an absorption filter
55
. Light from the light source
51
, which is a mercury lamp, is led to the excitation filter
53
through the illumination lens system
52
. Of the light from the light source
51
, only excitation light of wavelength needed to excite a sample
47
is selectively transmitted by the excitation filter
53
. Excitation light emanating from the excitation filter
53
is reflected by the dichroic mirror
54
and applied to the sample
47
through an objective
41
. Fluorescent light from the sample
47
is collected by the objective
41
and passes through the dichroic mirror
54
to reach the absorption filter
55
. The absorption filter
55
transmits only fluorescent light of specific wavelength selected according to the spectral characteristics thereof. The fluorescent light of specific wavelength is imaged through an imaging lens
43
and viewed as a fluorescence image through an eyepiece
44
. The fluorescence illumination optical system shown in
FIG. 43
projects an image of the light source
51
in the vicinity of the pupil position of the variable magnification optical system
42
L and allows the illumination area and the observation area to coincide with each other independently of a change in magnification made during observation and also independently of interchanging the objective
41
with another objective. Therefore, the fluorescence illumination optical system is excellent in operability.
Similarly, the fluorescence illumination optical system shown in
FIG. 44
projects an image of the light source
51
in the vicinity of the pupil position of the objective
41
and is therefore capable of making the illumination area and the observation area coincident with each other independently of interchanging the objective
41
with another objective.
FIG. 45
shows an arrangement in which an observation optical path is not used also as an illumination optical path, unlike the illumination method shown in FIG.
43
. The fluorescence illumination optical system shown in
FIG. 45
has a light source
51
, a collector lens system
58
, a light guide fiber
59
, an excitation filter
53
, and an illumination lens system
57
capable of varying the illumination area. Excitation light from the light source
51
is collected by the collector lens system
58
and led to an entrance end surface
59
a
of the light guide fiber
59
. Light emerging from an exit end surface
59
b
of the light guide fiber
59
passes through the illumination lens system
57
, which is capable of varying the illumination area, and further passes through the excitation filter
53
whereby only light in a specific wavelength region is selected and applied to a sample
47
. Fluorescent light from the sample
47
is viewed through an objective
41
, variable magnification optical systems
42
R and
42
L, absorption filters
55
R and
55
L, imaging lenses
43
R and
43
L and eyepieces
44
R and
44
L as in the case of FIG.
43
.
FIG. 46
shows the arrangement of an apparatus proposed in WO99/13370, in which a variable magnification optical system in
Kawasaki Kenji
Nishiwaki Daisuke
Shimizu Keiji
Fineman Lee
Olympus Corporation
Pillsbury & Winthrop LLP
Robinson Mark A.
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