Optical image separation system and confocal scanning unit

Optical: systems and elements – Compound lens system – Microscope

Reexamination Certificate

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C359S234000

Reexamination Certificate

active

06631029

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of Invention
This invention relates to a confocal observation system; and more particularly, to an optical image separation system connected to a confocal image output port of a Nipkow disk type confocal scanner.
2. Description of the Prior Art
FIG. 1
shows a conventional Nipkow disk type confocal scanner used with a microscope, wherein confocal scanner
100
is connected to microscope
200
. Illuminating parallel excitation light beam
1
(depicted as a chain line) is converged into individual beams by micro lens array disk (called “ML disk”)
2
, and passed through pinholes of pinhole array disk (called “Nipkow disk”)
4
after being transmitted through dichroic mirror (called “DM”)
3
. Light beam
1
is then focused onto sample
6
by objective lens
5
of the microscope
200
. Fluorescence signal
7
(depicted as a continuous line) emitted from sample
6
is transmitted through objective lens
5
and is focused on the pinholes of Nipkow disk
4
. Fluorescence signal
7
, passed through the pinholes, is reflected by DM
3
and is caused to form a fluorescence image on a two dimensional sensor
10
by relay lens
9
. DM
3
is designed to transmit excitation light
1
and to reflect the desired fluorescence signal
7
.
ML disk
2
, and Nipkow disk
4
are concurrently turned by rotation shaft
11
, both being mechanically connected to each other by member
8
. Mico lenses and pinholes formed on Nipkow disk
4
are arranged so that the pinholes scan over plane
12
to observe sample
6
. The plane on which pinholes of Nipkow disk
4
are arranged, plane
12
to be observed on sample
6
, and light detecting surface of the sensor
10
are arranged in optically conjugate relationship to each other. Accordingly, an optical sectional image of sample
6
, that is, a confocal image is formed on sensor
10
.
A plurality of specimens placed on the same plane can be simultaneously observed because a Nipkow disk type confocal scanner uses a two dimensional sensor. Also, a confocal image of sample
6
can be formed on the light detecting surface of the sensor
10
in a short time by rotating ML disk
2
and Nipkow disk
4
together at a high speed. This makes it possible to acquire at high speeds confocal images of all of the samples
60
, on which many specimens to be inspected are arranged in a matrix as shown in
FIG. 2
, moving them relative to the microscope and the confocal scanner. Accordingly, confocal scanners are used for screening procedures used in developing new medicine. Furthermore, when a sample emitting sufficiently bright fluorescence signals is used as the object of test or inspection, use of the ML disk
2
is sometimes not necessary.
The conventional confocal scanner has a problem in that the number of samples that can be processed per unit time in the screening procedure of
FIG. 2
, is limited to those where a sample emits only one type of fluorescence. Disadvantageously, when a multiple staining procedure is used, a plurality of different fluorescence wavelengths, depending on sample reactions to the different wavelengths, are detected. This is done by adding separate fluorescence reagents having different wavelengths of fluorescence for each sample
60
, or by adding a plurality of types of fluorescence reagents to all of the samples. In that case, sample
60
having a plurality of different specimen reacting to different fluorescence reagents, is first inspected using a DM
3
that has a specific reflection characteristic and can detect that specific first fluorescence wavelength. Then, after replacing the DM
3
with another DM
3
having another reflection characteristic corresponding to the second fluorescence wavelength, the entire specimen having the plurality of samples, must again be inspected. As can be appreciated, the conventional confocal scanner can be improved.
As shown in
FIG. 3
, one way of handling the foregoing problem is to use a plurality of sets of microscopes
200
a
,
200
b
,
200
c
; confocal scanners
100
a
,
100
b
,
100
c
; and two dimensional sensors
10
a
,
10
b
,
10
c
, arranged, respectively, for each fluorescence wavelength. When inspection for fluorescence wavelength &lgr;n to sample
60
n
is completed for one wavelength, sample
60
n
is sent to the next inspections system n+
1
comprising microscope
200
n
+
1
, scanner
100
n
+
1
and two dimensional sensor
10
n
+
1
, and then the process is repeated for the next fluorescence wavelength of the sample. In other words, the inspection for each fluorescence wavelength is repeated using the different sets. However, this arrangement is not satisfactory since it involves use of a plurality of sets of devices and hence cost is expensive, and repeated procedures are time consuming.
SUMMARY OF THE INVENTION
Accordingly, an object of the invention is to overcome the aforementioned and other deficiencies, disadvantages and problems of the prior art.
Another object is to provide an optical image separation system connected to a Nipkow disk type confocal scanner, wherein return light from a sample is detected by separating the return light concurrently into a plurality of different wavelength regions without repeating the DM changes, as done in the prior art; and inspection of the total number of specimen by using the entire number of wavelengths, without using a plurality of sets of scanners and microscopes, as done in the prior art.
The foregoing and other objects are attained in the invention which encompasses an optical image separation system connected to a confocal image output port of a Nipkow disk type confocal scanner, wherein the optical image separation system comprises a return light separating means that separates the light returned from a sample and emitted from the output port of the scanner into light beams of a plurality of wavelength regions or a plurality of portions of the same wavelength region.
The various features, aspects, and advantages and effects of the invention are further detailed hereinbelow.


REFERENCES:
patent: 5717519 (1998-02-01), Sugiyama et al.
patent: 5969846 (1999-10-01), Kishi
patent: 6297904 (2001-10-01), Kitigawa et al.
patent: 6426835 (2002-07-01), Endo et al.

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