Inverted confocal microscope

Details Inverted confocal microscope Inverted confocal microscope

1999-09-21

2001-10-02

Sikder, Mohammad (Department: 2872)

Optical: systems and elements

Compound lens system

Microscope

C359S369000, C359S370000, C359S372000

Reexamination Certificate

active

06297904

ABSTRACT:

CROSS REFERENCE TO THE RELATED APPLICATION
Priority is claimed for the present application based on Japanese Patent Application Nos. 98/268338 (filing date: Sep. 22, 1998), 98/268312 (filing date: Sep. 22, 1998) and 98/268313 (filing date: Sep. 22, 1998), the entire contents of which are incorporated herein by reference.
BACKGROUND OF THE INVENTION
The present invention relates to an inverted confocal microscope used for the observation of biological sample as well as for metallic and industrial purposes.
An inverted confocal microscope having an objective lens arranged below the bottom of a culture container, such as a petri dish and a flask, has been conventionally used as a microscope for observing, measuring and imaging the dynamic behavior of a biological sample such as a cultured cell.
The inverted confocal microscope of this type for the study of cell biology involves one disclosed by Jpn. Pat. Appln. KOKAI Publication No. 7-035986. This microscope is designed to have a vertical fluorescent light projection tube for observing a fluorescent light arranged so that the fluorescent image of a live cell into which a fluorescent indicator such as for Ca ions or pH is injected. This microscope includes therein an optical path branching optical system to serve a plurality of image pick-up means such as a photographing still camera, a CCD camera having high space resolution and a high-speed image pick-up device having high time resolution in addition to a device for the observation of a sample, and obtains a plurality of image pick-up optical paths besides an observation optical path.
Meanwhile, as an inverted microscope to provide an image of a live cell at high speed, there is disclosed a microscope in which a confocal scanner unit is detachably attached to a microscope main body by Jpn. Pat. Appln. KOKAI Publication No. 5-60980. The microscope disclosed by 5-60980 employs a confocal disk scanner as a confocal scanner unit. The confocal disk used for confocal disk scanner is constructed by coupling a light condensing disk having multiple pinhole openings into which a plurality of microlenses are embedded, to a pinhole disk having a plurality of pinholes formed to correspond to the multiple pinholes.
Here, each of the pinholes formed in the pinhole disk transmits both an illumination light to a sample and a fluorescent light from the sample, and forms a confocal optical system. With this constitution, the optical disconnect effect of the confocal microscope allows obtaining a high-contrast, clear image. In addition, since the microscope uses a rotary disk scanner, it can observe and pick up an image in a real time manner. It is, in particular, desirable to use this microscope for observing and measuring the physiologically dynamic behavior of a cell by picking up the fluorescent image of a live cell into which a fluorescent indicator for, for example, Ca ions and pH is injected, at high speed.
By the way, the confocal scanner unit as stated above is attached to a microscope main body using a C mount adapter. As a specific way to attach the confocal scanner unit to an inverted microscope is, as shown in, for example,
FIG. 1A
, the confocal scanner unit is attached to one of three optical takeout ports including a straight barrel
101
on an observation lens barrel
100
, a left (or right) side port
103
of a lens barrel
102
and a bottom port
104
below the lens barrel
102
. Namely, the confocal scanner unit is attached to one of the three ports and a still camera or a small size CCD camera is attached the remaining two ports.
FIG. 1B
shows a state in which the confocal scanner unit
105
is attached to the respective optical path takeout ports through the C mount adapter.
However, if the confocal scanner unit
105
is attached to one of the optical path output ports stated above through the C mount adapter or the like, the following disadvantages occur.
First, in case of attaching the confocal scanner unit
105
to the straight barrel
101
above the observation lens barrel
100
, an observer's eye point is higher than that during ordinary observation and only a monocular lens can be disadvantageously used. Due to this, at the time of observing a confocal image macroscopically, not only an overstrain affects the observer's posture but also it becomes difficult for the observer to operate the microscope and handle the sample during observation.
In case of attaching the confocal scanner unit
105
to the side port
103
on the left (or right) of the lens barrel
102
, the confocal scanner unit
105
is attached at a lower position of the side of the lens barrel, with the result that operability deteriorates. In case of macroscopically observing a confocal image with an eye piece fixed to the confocal scanner unit, in particular, the observer is forced to endure an overstraining posture.
In case of attaching the confocal scanner unit
105
to the bottom port
104
below the lens body
102
, the confocal scanner is attached onto the bottom of the lens barrel, i.e., the back side of a table on which the microscope is mounted and the operability of the confocal scanner unit is considerably hampered. The macroscopic observation of a confocal image with the confocal scanner unit is particularly quite difficult to make.
BRIEF SUMMARY OF THE INVENTION
The present invention has been made under the above-stated circumstances. It is, therefore, an object of the present invention to provide an inverted confocal microscope which enables comfortably making macroscopic confocal observation, which has excellent operability and which enables easily comparing a confocal image with a non-confocal image on an observation optical path and a image pick-up optical path.
To attain the above object, according to the main scope of the present invention, there is provided an inverted microscope that reflects an observation light passing through an image-formation optical system upward using at least one reflection light optical system and that leads the observation light to an observation optical path, said inverted microscope comprising:
a confocal scanner arranged at an image surface position of the image-formation optical system;
a light source for applying a light onto a sample through the confocal scanner and the image-formation optical system;
a confocal image formation optical system leading the light passing through the confocal scanner to the sample through the image-formation optical system, returns a return light from the sample to the confocal scanner along a route opposite to that for leading the light to the sample, and obtains a confocal image; and
a confocal observation optical system for leading the return light from the sample passing through the confocal scanner to the observation optical path.
With this constitution, even if the confocal scanner is attached to the inverted microscope, an observer can observe a confocal image without changing the height of the observer's eye point.
According to one embodiment, the confocal image formation optical system is provided between the image-formation optical system and a reflection optical system to be freely insertable and detachable and includes a first optical path branching optical system forming a branching optical path; and
the confocal scanner is provided on a branching optical path formed by the first optical path branching optical system.
According to one embodiment, the confocal observation optical system comprises:
a second optical path branching optical system provided at the confocal scanner and leading a light passing through the confocal scanner from the scanner to the observation optical system; and
a third optical path branching optical system putting the light branched by the second optical system on the observation optical path.
According to one embodiment, the frequency of reflection of light by the first to third optical path branching optical systems is set at even-number times.
According to one embodiment, the confocal scanner is provided on an observation optical path formed by the reflection optica

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