CN1300563C - Minisize three-dimensional self-scanning confocal microscope - Google Patents
Minisize three-dimensional self-scanning confocal microscope Download PDFInfo
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- CN1300563C CN1300563C CNB2005100184294A CN200510018429A CN1300563C CN 1300563 C CN1300563 C CN 1300563C CN B2005100184294 A CNB2005100184294 A CN B2005100184294A CN 200510018429 A CN200510018429 A CN 200510018429A CN 1300563 C CN1300563 C CN 1300563C
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Abstract
The present invention discloses a miniature three-dimensional self-scanning confocal microscope. A double-comb linear scanner is composed of an actor, an X direction linear driver and a Z direction linear driver which are used as two stators, wherein a second microlens is arranged on the actor; light emitted by a spot type light source becomes a collimated laser beam after passing through a first microlens; after the collimated laser beam penetrates through a miniature optical beam splitter and is reflected by a micro scanning reflector, the beam is focused on a sample by the second microlens; after reflected light or fluorescent light is orderly reflected by the second microlens and the micro scanning reflector along an original optical path, the light returns to the miniature optical beam splitter; the light reflected by the miniature optical beam splitter is irradiated to a miniature optical detector through a third microlens and a pin hole; a control device is used for data acquisition and processing, and controls the double-comb linear scanner to scan in the linear of focal points in X and Y directions. The present invention has the characteristics of small volume, simple structure, good stability, high frequency response of dynamic characteristics and direct three-dimensional solid self-scanning measurement.
Description
Technical field
The invention belongs to the detection technique field, be specifically related to a kind of minisize three-dimensional self-scanning confocal microscope.
Background technology
Confocal microscope obtains widespread use as a kind of strong imaging analysis instrument of translucent biological specimen (comprising cell and tissue) in biomedical research.In addition, it also is widely used in the non-cpntact measurement and the research of microfabrication and various engineering surfaces, and the vision of autonomous Micro-Robot.Scholars have carried out big quantity research to this technology in recent years, the main all the time confocal microscope based on routine.
(see Rep.Prog.Phys.1996,59:427-471 at " Confocal optical microscopy ".) confocal microscope of laser simple scan mode or dish formula conjugation dot matrix scan mode proposed in the literary composition.Its three-dimensional measurement technology normally adopts two-dimensional layer area scanner structure to add one dimension vertical feed mechanism, realizes the measurement of a plurality of two-dimentional aspects, and then the reconstruct three-dimensional feature.This measuring technique effect is remarkable, but the frequency response big, dynamic perfromance of its structure is low, has limited its application, has particularly hindered it and has been directly used in the biosome.And the scanning mechanism of the method is comparatively complicated, and the influence of vibration has limited the raising of measuring accuracy and speed, the incompatible demand of quick online detection at present.
" Stacked two dimensional micro-lens scanner for micro confocalimaging array. " (sees [2] Micro Electro Mechanical Systems, 2002.TheFifteenth IEEE International Conference:483-486) the micro scanning lens mechanism of two-dimentional linear drives, location is used for the array of confocal scan microscope, has realized two-dimensional scan.
Summary of the invention
The objective of the invention is to overcome above-mentioned the deficiencies in the prior art part, a kind of minisize three-dimensional self-scanning confocal microscope is provided.This minisize three-dimensional self-scanning confocal microscope has the advantages that volume is little, the frequency response of dynamic perfromance high and direct 3 D stereo self-scanning is measured.
A kind of minisize three-dimensional self-scanning confocal microscope provided by the invention comprises point type light source, three lenticules, micro-optic beam splitter, micro scanning catoptron, two pectination rectilinear scanner, pin hole, miniature photo-detector and control device; Wherein, two pectination rectilinear scanners are made up of to linear actuator to linear actuator and Z mover and two X as stator, second lenticule is positioned on the mover, X to linear actuator be used to drive mover along X to moving, Z to linear actuator be used to drive mover along Z to moving; The light that the point type light source sends through first lenticule after, become collimated laser beam; Collimated laser beam sees through the micro-optic beam splitter, after the micro scanning mirror reflects, focuses on sample by second lenticule; Reflected light or fluorescence along original optical path successively through after second lenticule and the micro scanning mirror reflects, turn back to the micro-optic beam splitter, the light that is reflected by the micro-optic beam splitter shines miniature photo-detector through the 3rd lenticule and pin hole, is converted into photosignal by it and flows to control device; The scanning of control device control micro scanning mirror deflection forms the focus scanning of Y direction on the second lenticular focal plane; X is to linear actuator in control device control, drives clamping and lenticular mover, forms the focus rectilinear scanning of directions X; Z is to linear actuator in control device control, drives clamping and lenticular mover, forms the focus rectilinear scanning of Z direction; Control device carries out obtaining after the acquisition process position signalling of three directions of the correspondence of these spatial point to the 3-D scanning signal; Miniature photo-detector obtains the light intensity signal of these corresponding spatial point, and sends control device to, and control device obtains the set of light intensity signal and relative position signals thereof.
The present invention adopts a micro scanning catoptron and has the two pectination rectilinear scanners and the lenticular object lens of driving that move along mutually perpendicular direction and realized direct 3 D stereo self-scanning.Simultaneously, the present invention utilizes the technology of MEMS (micro electro mechanical system) (MEMS), on the basis that keeps confocal microscope high measurement resolution and non-cpntact measurement advantage, compact integrated with MEMS devices such as lasing light emitter, micro-optical device, micro scanner and miniature photo-detectors, have that volume is little, the dynamic perfromance frequency response is high and direct characteristics such as 3 D stereo self-scanning measurement, can solve with this that conventional confocal microscope structure is big, the low problem of dynamic perfromance frequency response; That the present invention also has is simple in structure, make the characteristics of easy and good stability.
Compare with the two-dimensional scan method of double micro lens, this minisize three-dimensional self-scanning confocal microscope not only has the three-dimensional self-scanning ability, and have light path weak point, characteristic of simple, and then can under the minimal noise condition miniature three-dimensional scanning be integrated in the 2mm level size from the confocal microscope explorer portion.Because the quality of movement parts is very little, the working limit frequency of its 3-D scanning is greater than 1kHz.
Description of drawings
Fig. 1 is the structural representation of minisize three-dimensional self-scanning confocal microscope.
Fig. 2 is the control principle synoptic diagram of minisize three-dimensional self-scanning confocal microscope.
Embodiment
Be described in further detail for the present invention below in conjunction with accompanying drawing.
As shown in Figure 1, minisize three-dimensional self-scanning confocal microscope comprises point type light source 1, first lenticule 2, micro-optic beam splitter 3, minisize three-dimensional self-scanning device, the 3rd lenticule 10, pin hole 11, miniature photo-detector 12 and control device 13.
The minisize three-dimensional self-scanning device comprises micro scanning catoptron 4, second lenticule 5 and two pectination rectilinear scanner.Wherein, two pectination rectilinear scanners are made up of to linear actuator 8 to linear actuator 7 and Z a mover 6 and two X as stator, second lenticule 5 is positioned on the mover 6, X to linear actuator 7 be used to drive mover 6 along X to moving, Z to linear actuator 8 be used to drive mover 6 along Z to moving.
Point type light source 1 can be miniature point type laser instrument or miniature point type laser diode.
The light that point type light source 1 sends through first lenticule 2 after, become collimated laser beam.Collimated laser beam sees through micro-optic beam splitter 3, after 4 reflections of micro scanning catoptron, focuses on sample 9 by second lenticule 5.Reflected light or fluorescence along original optical path through 4 reflections of second lenticule 5 and micro scanning catoptron after, turn back to micro-optic beam splitter 3.By the light of micro-optic beam splitter 3 reflection through the 3rd lenticule 10 and pin hole 11, after the influence by the 3rd lenticule 10 and pin hole 11 elimination out of focus light, shine miniature photo-detector 12, be converted into photosignal and flow to control device 13, thereby obtain the light signal of a spatial point, the signal of this light intensity is height localization (corresponding spatial point).
The result that the 3 d grid formula stereoscanning of a microchannel is measured is the optical information of 3 D stereo dot matrix, and 3 D stereo is about 40*40*40 μ m.The working limit frequency of its 3-D scanning is greater than 1kHz
Remainder among Fig. 1 except that control device 13 can be referred to as the minisize three-dimensional self-scanning confocal microscope detector.It can form the array of minisize three-dimensional self-scanning confocal microscope detector, so that increase work efficiency.
Point type light source 1 adopts the point type semiconductor laser of power μ W level, and the size of its luminous zone is less than 5 μ m.Micro-optic beam splitter 3 adopts polysilicon to make, and the mirror of micro scanning catoptron 4 adds the aluminium film by polysilicon and makes; Lenticule is made by polymer, and its focal length is less than 1.0mm, and diameter is less than 0.5mm.
The present invention is not limited to example recited above, and persons skilled in the art can adopt numerous embodiments to realize the present invention according to content disclosed by the invention.
Claims (1)
1, a kind of minisize three-dimensional self-scanning confocal microscope is characterized in that: comprise point type light source (1), first lenticule (2), micro-optic beam splitter (3), micro scanning catoptron (4), second lenticule (5), two pectination rectilinear scanner, the 3rd lenticule (10), pin hole (11), miniature photo-detector (12) and control device (13); Wherein,
Two pectination rectilinear scanners are made up of to linear actuator (8) to linear actuator (7) and Z mover (6) and two X as stator, second lenticule (5) is positioned on the mover (6), X to linear actuator (7) be used to drive mover (6) along X to moving, Z to linear actuator (8) be used to drive mover (6) along Z to moving;
The light that point type light source (1) sends through first lenticule (2) after, become collimated laser beam; Collimated laser beam sees through micro-optic beam splitter (3), after micro scanning catoptron (4) reflection, focuses on sample (9) by second lenticule (5); Reflected light or fluorescence along original optical path successively through after the reflection of second lenticule (5) and micro scanning catoptron (4), turn back to micro-optic beam splitter (3), by the light of micro-optic beam splitter (3) reflection through the 3rd lenticule (10) and pin hole (11), shine miniature photo-detector (12), be converted into photosignal by it and flow to control device (13);
Control device (13) control micro scanning catoptron (4) deflection scanning forms the focus scanning of Y direction on the second lenticular focal plane; X is to linear actuator (7) in control device (13) control, drives clamping and the mover (6) of lenticule (5), forms the focus rectilinear scanning of directions X; Z is to linear actuator (8) in control device (13) control, drives clamping and the mover (6) of lenticule (5), forms the focus rectilinear scanning of Z direction; Control device (13) carries out obtaining after the acquisition process position signalling of three directions of the correspondence of these spatial point to the 3-D scanning signal; Miniature photo-detector (12) obtains the light intensity signal of these corresponding spatial point, and sends control device to, and control device (13) obtains the set of light intensity signal and relative position signals thereof.
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US10908403B2 (en) | 2011-02-14 | 2021-02-02 | European Molecular Biology Laboratory (Embl) | Light-pad microscope for high-resolution 3D fluorescence imaging and 2D fluctuation spectroscopy |
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WO2003107064A1 (en) * | 2002-06-01 | 2003-12-24 | オリンパス光学工業株式会社 | Confocal microscope and method for measuring by confocal microscope |
JP2004279342A (en) * | 2003-03-18 | 2004-10-07 | Jasco Corp | Apparatus for measuring depth |
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US6122098A (en) * | 1996-06-11 | 2000-09-19 | Evotec Biosystems A.G. | Confocal microscope for optical determination of an observation volume |
CN2416510Y (en) * | 2000-04-25 | 2001-01-24 | 南京理工大学 | Laser confocal screening microscope |
WO2003036227A1 (en) * | 2001-10-25 | 2003-05-01 | Camtek Ltd. | Confocal wafer-inspection system and method |
WO2003107064A1 (en) * | 2002-06-01 | 2003-12-24 | オリンパス光学工業株式会社 | Confocal microscope and method for measuring by confocal microscope |
JP2004279342A (en) * | 2003-03-18 | 2004-10-07 | Jasco Corp | Apparatus for measuring depth |
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US10908403B2 (en) | 2011-02-14 | 2021-02-02 | European Molecular Biology Laboratory (Embl) | Light-pad microscope for high-resolution 3D fluorescence imaging and 2D fluctuation spectroscopy |
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