CN104865688B - The microscope of external fluorescent moieties - Google Patents
The microscope of external fluorescent moieties Download PDFInfo
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- CN104865688B CN104865688B CN201510338469.0A CN201510338469A CN104865688B CN 104865688 B CN104865688 B CN 104865688B CN 201510338469 A CN201510338469 A CN 201510338469A CN 104865688 B CN104865688 B CN 104865688B
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/06—Means for illuminating specimens
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Abstract
The invention provides a kind of microscope of Novel external fluorescent moieties, its lighting device, comprise the fluorescent moieties of an external light source and a filter set formation.Described fluorescent moieties is placed between object lens and sample, is positioned at immediately below object lens.Microscopical source path is followed successively by external light source, fluorescent moieties, sample, fluorescent moieties, object lens, eyepiece.The microscope of Novel external fluorescent moieties of the present invention is compared with traditional microscope, and light loss is very little, can obtain that hot spot is large, light intensity is large, the specimen illumination of homogeneous and bright.Be particularly useful for space required large, the bio signal detection that fluorescence signal is weak.
Description
Technical field
The present invention relates to a kind of microscope, particularly a kind of microscope of Novel external fluorescent moieties.
Background technology
The fluorescent moieties of current fluorescent microscope is all between object lens and eyepiece, adopt mercury lamp, xenon lamp or Halogen lamp LED as light source, exciting light arrives colour filter block group (colour filter block group is made up of to color separation film, transmitting optical filter exciter filter, two) by cola illuminator, light source obtains required excitation spectrum through exciter filter, be incident upon sample on to color separation film reflection through object lens through two, after being excited by the sample of fluorescent dyeing, emission wavelength is greater than the spectrum of exciting light.Utilizing emitted light arrives eyepiece or imaging device through object lens, two to color separation film, transmitting optical filter (beyond utilizing emitted light, the spectrum of wave band is cut off).
At present, publication number be CN104035195A patent discloses a kind of fluorescent microscope, comprise support body, objective table and base, described support body, objective table and base set gradually from top to bottom, the all sides of described support body are provided with more than one adjusting knob, described support body bottom is provided with object lens, described objective table is positioned at position immediately below object lens, described objective table bottom position is disposed with condenser, diaphragm and fixed support, medium position on described base is provided with LED fluorescence excitation light source, described LED fluorescence excitation light source is just to the medium position of objective table, more than one reflective mirror swivel mount is provided with around described LED fluorescence excitation light source, described LED fluorescence excitation light source includes lampshade, described lampshade is provided with radome, described radome is flexibly connected with LED fluorescence excitation light source, each reflective mirror swivel mount described is all provided with reflective mirror, this fluorescent microscope strengthens fluorescent effect by being provided with multiple concentrating device.Publication number be CN202735583A patent discloses a kind of fluorescent microscope, comprise object lens, object lens are arranged on object lens converter, object lens converter is connected with eyepiece stalk, eyepiece stalk is arranged on handel, handel is arranged on fixed station, objective table is connected with fixed station, sample is installed above objective table and advances chi, sample advances chi to advance chi spiral to be connected with sample, below objective table, condenser is installed, condenser is connected with diaphragm, fixed station is provided with coarse adjustment knob and fine adjustment knob, fixed station is arranged on microscope base, described microscope base is provided with LED fluorescence excitation light source, its lighting system is for falling to penetrating formula, namely light source will be projeced on sample by object lens, but light loss can be caused like this to strengthen.
The fluorescent moieties of conventional microscope is between micro objective and imaging len, and because of the impact of diaphragm, various lens and object lens, the hot spot light loss obtained is very large, sometimes reaches 90%, cause light intensity not or light source visual field little.
But for some Bioexperiment, need the uniform sample illumination of large area, light field.As digital pcr, its sample, for including several ten thousand even up to a million parts of reaction members, will carry out observation and analysis to these reaction members simultaneously, and sample illumination must meet following three conditions simultaneously: (1) Large visual angle light source, (2) light intensity is large, (3) uniform intensity.Meet observation and analysis while that these three conditions just can being carried out the sample of the reaction member of these large quantity, especially area is large, and light intensity is greatly the factor of overriding concern simultaneously.And traditional lighting device visual field is very little, this demand cannot be met; Although traditional microscope can increase sample visual field by adopting the object lens of low range, the intensity of the sample illumination obtained like this reduces with the second power of the increase multiple of visual field, cannot meet the needs that digital pcr is analyzed equally.Thus us are impelled to develop a kind of microscope illuminator of Novel external fluorescent moieties.
Summary of the invention
The object of the invention is for solving the problems such as conventional microscope light loss is large, light source visual field is little, light intensity is not enough, providing that a kind of light source visual field is large, light intensity large, the lighting device of uniform intensity, its technical scheme is as follows:
The microscope of Novel external fluorescent moieties, its lighting device comprises the fluorescent moieties of an external light source and filter set formation.
Described fluorescent moieties is placed between object lens and sample, is positioned at immediately below object lens.
External light source is selected from the one in mercury lamp, Halogen lamp LED, xenon lamp or LED.
When external light source is LED, the microscope of described Novel external fluorescent moieties also comprises light collecting device.
Described filter set is made up of to color separation film, transmitting optical filter exciter filter, two.Exciter filter with transmitting optical filter become 90 degree vertical, two to color separation film divide equally exciter filter with transmitting optical filter formed by right angle, two become miter angle to color separation film with microscopical optical axis.
The effect of exciter filter is by exciting light filtering, obtains narrowband excitation wave band, avoids the long-wave band light in light source to enter filter set.Two is reflected by short-wavelength light to the effect of color separation film, long wavelength light transmission.Launching optical filter is by the long wavelength light transmission of fluorescent emission, filters the light of short wavelength.
Described filter set adopts the optical filter of different spectral characteristic for different fluorophor.When external light source is LED, the now LED of the corresponding different-waveband of the optical filter of different spectral characteristic.
The microscopical source path of Novel external fluorescent moieties of the present invention is followed successively by external light source, fluorescent moieties, sample, fluorescent moieties, object lens, eyepiece.Compare traditional microscope, the microscope light loss of Novel external fluorescent moieties of the present invention is very little, the facula area obtained and light intensity large.
The present invention is particularly useful for needing sample visual field large, the bio signal detection that fluorescence signal is weak.
The lighting device of external fluorescent moieties provided by the invention has the following advantages:
1, facula area is large, and traditional microscope, facula area is no more than 0.5cm
2, and microscopes of the present invention produces at least 2cm
2hot spot, also customizable fluorescent moieties, produce 9cm
2above hot spot.
2, light loss is little, and light intensity is large, and fluorescent moieties is placed between object lens and sample by microscope of the present invention, and its light loss is only 30%, compare conventional microscope up to 90% light loss, microscope of the present invention effectively can improve the intensity of light source.
3, as adopted LED to also have that power is low, price is low, the life-span is long as external light source, start shooting without the need to preheating, shut down without the need to advantages such as coolings.
Accompanying drawing explanation
Fig. 1 is the microscopical structural drawing of Novel external fluorescent moieties of the present invention.
Fig. 2 is the microscopical light path process flow diagram of Novel external fluorescent moieties of the present invention.
Fig. 3 is the structural drawing of conventional microscope.
Fig. 4 is the light path process flow diagram of conventional microscope.
Embodiment
Now the invention will be further described in conjunction with the accompanying drawings and embodiments.
Embodiment 1
Shown in seeing figures.1.and.2, fluorescent moieties 1 comprises exciter filter 11, two and forms to color separation film 12, transmitting optical filter 13.Fluorescent moieties is placed between object lens 3 and sample 2, be positioned at immediately below object lens 3, exciter filter 11 is vertical with transmitting optical filter 13 one-tenth 90 degree, and two divide exciter filter 11 and right angle formed by transmitting optical filter 13 equally to color separation film 12, and two become miter angle to color separation film 12 with microscopical optical axis.
First, the light that external light source 5 sends enters fluorescent moieties 1, through exciter filter 11 filtering, obtain the light of specific band short wavelength, the light of this specific band short wavelength be mapped to become miter angle to place with microscopes optical axis two on color separation film 12, now two show reflection characteristic to color separation film 12, the light of this short wavelength is reflexed on sample 2, after sample 2 is excited, emission wavelength is greater than the spectrum of exciting light, and then through two to color separation film 12, through launching optical filter 13, received by object lens 3, then enter eyepiece 4 or sniffer.
The microscope light source path of external fluorescent moieties of the present invention is followed successively by external light source, fluorescent moieties, sample, fluorescent moieties, object lens, eyepiece.
Adopt the method for the present embodiment, owing to being placed between object lens 3 and sample 2 by fluorescent moieties, the light path making light source arrive sample shortens, and avoids the light loss that light source causes through the convergence effect of diaphragm, various lens and object lens, therefore, light loss can be dropped to minimum.When exciter filter 11, two is of a size of 3cm × 3cm to color separation film 12, transmitting optical filter 13, available facula area is 9cm
2, adopt the blue-ray LED of 3W, now light intensity reaches 20mW/cm
2above.
Embodiment 2
With reference to shown in Fig. 3 and Fig. 4, first, the light transmission diaphragm 6 that external light source 5 sends and lens 7, enter fluorescent moieties 1, through exciter filter 11 filtering, obtain the light of short wavelength, the light of short wavelength is mapped to two on color separation film 12, now two show reflection characteristic to color separation film, reflexed to by the light of short wavelength on object lens 3, be irradiated on sample 2 through object lens 3, the light that sample 2 inspires long wavelength enters fluorescent moieties 1 through object lens 3 again, through launching optical filter 13, then enter eyepiece 4 or sniffer.
The source path of conventional microscope is followed successively by external light source, fluorescent moieties, object lens, sample, object lens, fluorescent moieties, eyepiece.
The area of the hot spot of conventional microscope is general less than 1cm
2, its index path needs through multiple diaphragm and lens, the impact of object lens in addition, and the hot spot light loss obtained is very large, sometimes reaches 90%, and adopt the blue-ray LED of 3W, light intensity is only 1mW/cm
2.
Above to the microscope of the Novel external fluorescent moieties that the embodiment of the present invention provides, be described in detail, apply specific case herein to set forth principle of the present invention and embodiment, the explanation of above embodiment just understands method of the present invention and core concept thereof for helping; Meanwhile, for one of ordinary skill in the art, according to thought of the present invention, all will change in specific embodiments and applications, in sum, this description should not be construed as limitation of the present invention.
Claims (5)
1. a microscope for external fluorescent moieties, described microscopical lighting device comprises the fluorescent moieties of an external light source and filter set composition, it is characterized in that: described fluorescent moieties is placed between object lens and sample, is positioned at immediately below object lens;
Wherein, filter set is made up of to color separation film and transmitting optical filter exciter filter, two, exciter filter with transmitting optical filter become 90 degree vertical, two to color separation film divide equally exciter filter with transmitting optical filter formed by right angle, two become miter angle to color separation film with microscopical optical axis.
2. the microscope of external fluorescent moieties according to claim 1, is characterized in that: described microscopical source path is followed successively by external light source, fluorescent moieties, sample, fluorescent moieties, object lens, eyepiece.
3. the microscope of external fluorescent moieties according to claim 1, is characterized in that: described microscope also comprises light collecting device.
4. the microscope of external fluorescent moieties according to claim 1, is characterized in that: described filter set adopts the optical filter of different spectral characteristic for different fluorophor, the LED of the corresponding different-waveband of optical filter of different spectral characteristic.
5. the microscope of external fluorescent moieties according to claim 1, is characterized in that: described microscope is applicable to need sample visual field large, the bio signal detection that fluorescence signal is weak.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108226104A (en) * | 2016-12-22 | 2018-06-29 | 中国人民公安大学 | The LED phosphorimagers that a kind of long wave inhibits |
| CN106950686A (en) * | 2017-04-14 | 2017-07-14 | 苏州影睿光学科技有限公司 | A kind of near-infrared fluorescent inverted microscope system and its application |
| CN107271442A (en) * | 2017-07-21 | 2017-10-20 | 黄宁 | A kind of medical test multipurpose test system |
| CN110554009A (en) * | 2018-05-30 | 2019-12-10 | 宝山钢铁股份有限公司 | Magnetic powder inspection fluorescence imaging device for detecting steel profiled parts |
| CN110554085A (en) * | 2018-05-30 | 2019-12-10 | 宝山钢铁股份有限公司 | Integrated camera assembly for detecting magnetic powder inspection fluorescence imaging of small-sized special-shaped piece |
| CN109106326A (en) * | 2018-08-20 | 2019-01-01 | 广西奥顺仪器有限公司 | A kind of fluorescent apparatus for surgical operation microscope |
| CN112255207B (en) * | 2020-09-22 | 2023-12-12 | 广州源起健康科技有限公司 | Fluorescence excitation acquisition system |
| WO2023075695A2 (en) * | 2021-10-29 | 2023-05-04 | Agency For Science, Technology And Research | External illuminator, a microscope, and a microscopy method |
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| JP2014098835A (en) * | 2012-11-15 | 2014-05-29 | Canon Inc | Illumination optical system for microscope and microscope using the same |
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| CN1700051A (en) * | 2004-05-21 | 2005-11-23 | 株式会社其恩斯 | Fluorescence microscope |
| DE202007019607U1 (en) * | 2007-06-12 | 2014-04-01 | Carl Zeiss Microscopy Gmbh | Microscope for observing a sample in brightfield transmitted light or fluorescence incident light contrasting |
| CN102089418A (en) * | 2008-04-09 | 2011-06-08 | 耐克思乐生物科学 | Systems and methods for counting cells and biomolecules |
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Effective date of registration: 20180509 Address after: 116023 Liaoning high tech Industrial Park, Dalian, new high tech 1, E 2, 203-208 rooms. Patentee after: Dalian Yin Rui biomedical Instruments Co., Ltd. Address before: 100083 No. 35, Qinghua East Road, Beijing, Haidian District Patentee before: Semiconductor Inst., Chinese Academy of Sciences |
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Effective date of registration: 20190821 Address after: 315300 Room 801, Building 2, Science and Technology Plaza, Zhixi Wisdom Valley, Gutang Street, Cixi City, Ningbo City, Zhejiang Province Patentee after: Ningbo Xurui Biomedical Instruments Co., Ltd. Address before: Room 203-208, 2nd floor, Pioneer Port E, No. 1 Gaoxin Street, Dalian High-tech Industrial Park, Liaoning Province Patentee before: Dalian Yin Rui biomedical Instruments Co., Ltd. |