U.S. patent application number 11/976001 was filed with the patent office on 2008-04-17 for optical scanning microscope.
Invention is credited to Ralf WOLLESCHENSKY.
Application Number | 20080088920 11/976001 |
Document ID | / |
Family ID | 35134713 |
Filed Date | 2008-04-17 |
United States Patent
Application |
20080088920 |
Kind Code |
A1 |
WOLLESCHENSKY; Ralf |
April 17, 2008 |
Optical scanning microscope
Abstract
Microscope, in particular an optical scanning microscope with
illumination of a specimen via a beam splitter, which is arranged
in an objective pupil and includes at least a reflecting first
portion and at least a transmitting second portion, whereby the
reflecting portion serves to couple in the illumination light and
the transmitting portion serves to pass the detection light in the
detection direction or the transmitting portion serves to couple in
the illumination light and the reflecting portion serves to couple
out the detection light, with a first scanning arrangement. Means
are provided in the detection light path for the overlay of at
least one further scanning arrangement for illumination and
detection.
Inventors: |
WOLLESCHENSKY; Ralf;
(Apolda, DE) |
Correspondence
Address: |
JACOBSON HOLMAN PLLC
400 SEVENTH STREET N.W.
SUITE 600
WASHINGTON
DC
20004
US
|
Family ID: |
35134713 |
Appl. No.: |
11/976001 |
Filed: |
October 19, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10967321 |
Oct 19, 2004 |
7301696 |
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11976001 |
Oct 19, 2007 |
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Current U.S.
Class: |
359/385 |
Current CPC
Class: |
G02B 21/002 20130101;
G02B 21/0032 20130101 |
Class at
Publication: |
359/385 |
International
Class: |
G02B 21/06 20060101
G02B021/06 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 16, 2004 |
DE |
10 2004 034 983.5 |
Claims
1. An optical scanning microscope comprising: a source of
illumination light, an objective pupil, a beam splitter for
illuminating a specimen with the illumination light, the beam
splitter being arranged in the objective pupil and including at
least one reflecting first portion and at least one transmitting
second portion, wherein one of the first and second portion couples
in the illumination light and the other of the, first and second
portion passes detection light in a detection direction, a first
scanning arrangement for scanning the illumination light onto the
beam splitter, and at least one further scanning arrangement for
illumination and detection, provided in the detection light
path.
2. An optical scanning microscope comprising: a source of
illumination light, an objective pupil, a beam splitter for
illuminating a specimen with the illumination light, the beam
splitter being arranged in the objective pupil and including at
least one reflecting first portion and at least one transmitting
second portion, wherein one of the first and second portions
couples in the illumination light and the other of the first and
second portion passes detection light in a detection direction, and
wherein the beam splitter has a side facing towards the specimen
and a side facing away from the specimen, a first scanning
arrangement for scanning the illumination light onto the beam
splitter, a detection unit for detecting specimen light from the
first scanning arrangement, at least one further scanning
arrangement coupled in via the beam splitter, and returning means
for detouring of a part of the specimen light at the side of the
beam splitter facing away from the specimen and for then returning
the detoured specimen light toward the detection unit for detecting
specimen light from the first scanning arrangement.
3. An optical scanning microscope comprising: a source of
illumination light, an objective pupil, a beam splitter for
illuminating a specimen with the illumination light, the beam
splitter being arranged in the objective pupil and including at
least one reflecting first portion and at least one transmitting
second portion, wherein one of the first and second portion couples
in the illumination light and the other of the first and second
portion passes detection light in a detection direction, a first
scanning arrangement for scanning the illumination light onto the
beam splitter, the first scanning arrangement having at least one
scan mirror, the scan mirror having a reflecting rear side, at
least one further scanning arrangement coupled in via the beam
splitter, and returning means for directing at least one part of
the specimen light to the reflecting rear side of the scan mirror
for return in the detection direction.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present patent application is a divisional of
application Ser. No. 10/967,321, filed Oct. 19, 2004, which is
incorporated in its entirety by reference herein.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention is directed to a microscope, in
particular an optical scanning microscope with illumination of a
specimen via a beam splitter
[0004] 2. Related Art
[0005] In U.S. Pat. No. 6,888,148 among other things a beam
splitter is described for a line scanner.
[0006] In a line scanner the specimen is illuminated with a line
focus (e.g. along the X-coordinate), which is shifted in the
coordinate (Y) perpendicular to the line. For this the source of
light is linearly focused into an intermediate image plane of the
microscope mechanism by means of optics. By the focusing in Y
direction in the intermediate image, for example by a cylinder
lens, a linear and diffraction-limited distribution of intensity
arises along X on the specimen. With further optics the light is
focused into the pupil of the microscope arrangement. In the pupil
levels of the microscope arrangement a line focus results in each
case. The pupil levels and the scanner are conjugate to each other
and to the rear focal plane of the microscope arrangement, so that
the scanner can induce the linear and diffraction-limited focused
distribution of intensity perpendicular to this (Y-coordinate in
the specimen). The focusing into the specimen is made by scan
optics, the tube lens and the objective. Relay optics produces
conjugate pupil levels of the microscope arrangement. Due to the
kind of the specimen reciprocal effect e.g. during an excitation
for fluorescence or luminescence the light emitted from the
specimen is of small spatial coherency. That is each point excited
in the specimen radiates essentially independently of the
neighboring points as point emitter into all directions in space.
The optics, (e.g. a microscope objective) displays the individual
point emitters together with the tube lens TL into an intermediate
image plane ZB of the microscope mechanism, whereby the pupil P is
illuminated homogeneously (broken light path) by wave fronts that
are essentially incoherent to each other and of different
directions of propagation. In the pupil is the element which
separates the excitation light from the detection light. It is
constructed as described in DE.
SUMMARY OF THE INVENTION
[0007] The present invention is directed to a microscope, in
particular an optical scanning microscope with illumination of a
specimen via a beam splitter, which is arranged in an objective
pupil and consists of at least a reflecting first portion and at
least a transmitting second portion, whereby the reflecting portion
serves to couple in the illumination light and the transmitting
portion serves to pass the detection light in the detection
direction or the transmitting portion serves to couple in the
illumination light and the reflecting portion serves to couple out
the detection light, with a first scanning arrangement, whereby
means are provided in the detection light path for the overlay of
at least one further scanning arrangement for illumination and
detection.
BRIEF DESCRIPTION OF THE DRAWINGS
[0008] FIG. 1 is a schematic view of a first embodiment of an
optical scanning microscope in accordance with the present
invention.
[0009] FIG. 2 is a schematic view of a second embodiment of an
optical scanning microscope in accordance with the present
invention.
[0010] FIG. 3 is a schematic view of a third embodiment of an
optical scanning microscope in accordance with the present
invention.
[0011] FIG. 4 is a schematic view of a fourth embodiment of an
optical scanning microscope in accordance with the present
invention.
[0012] FIG. 5 is a schematic view of a fifth embodiment of an
optical scanning microscope in accordance with the present
invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0013] FIG. 1:
[0014] The light of a far field-source of light LQ is focused with
suitable optics for the production of an illumination line, for
example a cylinder lens ZL, linearly into one level that is
conjugate to the pupil P of the microscope objective O, in which
there is a developed beam splitter ST in accordance with U.S. Pat.
No. 6,888,148, which exhibits a narrow linear transmitter range,
over which the line is displayed via transmission optics L1, L2,
scan optics SO, tube lens TL and objective O into the specimen PR.
A scanner SC is arranged in a pupil P, that moves the line quickly
over the specimen in a scan direction perpendicular to the line
expansion.
[0015] The light (broken) coming from the specimen is returned by
the beam splitter reflecting up to the narrow transmitter range in
direction of detection via a replaceable filter F as well as
detection optics PO toward a detector DE1, in front of which a slit
diaphragm can be arranged.
[0016] FIG. 2:
[0017] Here exemplary sources of light LQ1, LQ2 are represented in
addition to the elements represented in FIG. 1 on cross ports,
which can result also from bypass of only one source of light,
whereby wavelength and intensity can be adjusted
advantageously.
[0018] By use of achromatic beam splitters the special advantage is
that the same wavelength can be used for both sources of light LQ1,
LQ2, which can be formed also by allocation in and of the same
source of light. The intermediate images ZB and ZB1 are levels
conjugate to each other. Furthermore, the pupil levels of the
microscope arrangement P are conjugate levels to each other. The
conjugate levels in each case are produced by the effect of the
optics lying between them in each case (those acting as relay
optics--light paths only schematically drawn).
[0019] LQ 2 can be for example a point scanner. The illumination
light of the point scanner can be used advantageously for the
purposeful manipulation (e.g. uncaging) on certain specimen
ranges.
[0020] The illumination light of LQ2 is faded after passage by
separate scan optics SO2 as well as a scanner SC1 (a X/Y scanner
favorable) over a usual dichroic color divider FT1 into the
detection light path of the line scanner and arrives over the
reflective range of the divider ST toward the specimen PR.
[0021] The reflecting range of the beam splitter ST is thus used
advantageously for the reflection of a further scan light path.
[0022] The light coming from the specimen arrives on the one hand
at the detector DE1 and on the other hand depending on
interpretation of the color divider FT1 also via a further color
divider FT2 toward a second detector DE2.
[0023] For example fluorescence light excited by LQ1 coming from
the specimen arrives during appropriate interpretation by FT1 on
the detector DE1 while reflected light of the point scanner LQ2
arrives on the detector DE2. Furthermore different fluorescence
wavelengths excited also by LQ1 and LQ2 can arrive on the different
detectors DE1 and DE2.
[0024] Since the light moved by the scanner SC1 is moved here
additionally by the scanner SC, the scanner SC1 must be controlled
in such a way that it compensates for the movement of the scanner
SC and additionally realizes a relative position for line
illumination.
[0025] That is simple to realize if scanner SC1 moves slower in
comparison to the scanner SC.
[0026] The fluorescence light induced by LQ2 can be also guided on
the line detector DE1.
[0027] Depending on the position of the scanner 2 the fluorescent
spot moves away over the line detector DE 1, i.e. the light is
separated by the scanner 2 toward DE1.
[0028] FIG. 3:
[0029] Here a cross port KS1 is provided, that can be a separate
module and is between a microscope stand S with tube lens and
objective, a first scan unit SC1 and a second scan unit SC 2.
[0030] SC 1 can contain the described line scanner and SC2 a point
scanner for scanning and/or manipulation.
[0031] SCI and SC 2 are couplable with KS1 at interfaces.
[0032] For this several intermediate images ZB that are conjugate
to each other are available in KS1 (via the optics L1, L2). The
conjugate levels in each case are produced by the effect of the
optics lying between them (light paths only schematic).
[0033] At the beam splitter ST1, which is developed analogous to
the beam splitter ST a line is focused on the specimen by the
transmitting range. It is attached in one pupil level of the
microscope arrangement.
[0034] For example with SC1 excited light such as fluorescence
light in the specimen is reflected downward at ST1 and arrives over
FT3, which is here constructed such that it lets this light portion
pass through, as well as over several reflectors RF onto the other
side of ST1. This light is diverted by ST1 toward the detector DE1
via ST.
[0035] The fluorescence light excited by the line scanner, which is
reflected at ST1 to the side, is thus brought advantageously in the
entire width back into the light path toward DE1.
[0036] Thus a further scanner SC2 can be reflected via FT3, whereby
by appropriate training of FT3, which can be replaceable, different
fluorescence wavelengths can arrive at DEl and/or DE2. The mode of
operation is similar to the one described above.
[0037] Contrary to FIG. 2 the scanners SC1 and SC2 can work here
advantageously independent of each other.
[0038] FIG. 4:
[0039] Here the light is not guided via reflectors RF on the back
side of the beam splitter ST1 as in FIG. 3 but on the back side of
the scanner SC3, which is here a mirror that can reflect on its
front and back sides and further guides with its back side the
specimen light (descanned) coming from the specimen and excited by
the line scanner (LQ1 arrives from above on the front side of the
scanner mirror) to the detector DE1. FT3 is constructed here in
such a way that it lets through the light intended for the detector
DE1 and only reflects the light intended for DE2.
[0040] Thereby again different fluorescences excited by the line
scanner and the point scanner can be detected advantageously at the
same time.
[0041] FIG. 5:
[0042] Here the light excited by the line scanner is not descanned
as in FIG. 4 but arrives via FT1 directly at a surface detector
(CCD matrix, gegatete camera), i.e. the linear light distribution
coming from the specimen runs in the direction of the scan via the
receiver surfaces, which records thereby a specimen image.
[0043] Further scan arrangements can also be reflected by cascading
(arrangement of further color dividers FT into a common light
path). The scan arrangements can be arbitrary image-giving
arrangements. Examples are the already mentioned point scanners,
scanners of point of resonance, Nipkow scanner, line scanners and
multi-point scanners. Furthermore, these can also be far-field
based microscope systems. It is advantageous here that they exhibit
an intermediate image plane as interface.
* * * * *