CN105785560B - Dark field microscope for cell culture on-line observation - Google Patents
Dark field microscope for cell culture on-line observation Download PDFInfo
- Publication number
- CN105785560B CN105785560B CN201610325563.7A CN201610325563A CN105785560B CN 105785560 B CN105785560 B CN 105785560B CN 201610325563 A CN201610325563 A CN 201610325563A CN 105785560 B CN105785560 B CN 105785560B
- Authority
- CN
- China
- Prior art keywords
- movable support
- unit
- pipe diameter
- objective lens
- dark field
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000004113 cell culture Methods 0.000 title claims abstract description 16
- 238000005286 illumination Methods 0.000 claims abstract description 15
- 238000006243 chemical reaction Methods 0.000 claims description 8
- 230000007246 mechanism Effects 0.000 claims description 6
- 230000005012 migration Effects 0.000 claims description 2
- 238000013508 migration Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 4
- 239000002245 particle Substances 0.000 description 3
- 230000008859 change Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/06—Means for illuminating specimens
- G02B21/08—Condensers
- G02B21/10—Condensers affording dark-field illumination
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/24—Base structure
Abstract
The invention provides a dark field microscope for cell culture online observation, which at least comprises: illumination element, movable support, support base, objective, photosensitive element, pipe diameter unit, AD converting unit, power module and control system, illumination element sets up the top at movable support, movable support sets up on supporting the base, the pipe diameter unit is connected in movable support lower part, objective setting is on the pipe diameter unit and corresponds the below at illumination element, photosensitive element sets up the light source signal that is used for gathering on the objective in the pipe diameter unit, AD converting unit connects photosensitive element, control system connects AD converting unit, power module adopts this device can realize direct, relapse, real-time observation to the observation target for the dark field microscope power supply that is used for cell culture on-line observation, need not the sample preparation.
Description
Technical Field
The invention relates to a dark field microscope, in particular to a dark field microscope for cell culture online observation.
Background
Dark field microscopes typically use special dark field condensers to deflect the illumination light from entering the objective, and only the scattered light from the sample enters the objective. Thus a bright image is obtained on a dark background, as opposed to dark field illumination, where the illuminating light directly reaches the imaging plane, known as bright field illumination.
The Tyndall phenomenon (Tyndall phenomenon) means that when a condensed light beam is incident into a sol, a luminous light column can be seen from the side surface of the light beam, and when light rays are irradiated on particles in the process of light propagation, if the particle diameter is slightly smaller than the incident light wavelength, light scattering occurs, each particle becomes a luminous point, and a light column can be seen from the side surface.
Dark field microscopes make use of the above principle. The special optical configuration of the darkfield microscope prevents incident illumination from being incident on the objective, but the sample is illuminated and emits reflected light. When the microscope is used for microscopic examination, the illumination light cannot be directly observed, the plane view field vertical to the optical axis is dark, a bright object image formed by scattered light and reflected light can be clearly seen on a dark background, and the object image and the background cause great contrast. The sample, within the field of view, is illuminated by the oblique light rays, which scatter and reflect light from the surface of various structures of the sample, seeing bright contours of many organelles, such as nuclei, mitochondria, vacuoles, and certain contents. In the case of dividing cells, spindle threads and chromosomes are visualized.
However, most of the dark field microscopes now have the following problems: firstly, under general conditions, the user must extract the sample earlier before observing the sample, then place on the slide glass, then place and observe under objective, and the operation is very loaded down with trivial details, is unfavorable for observing at any time of sample.
Disclosure of Invention
In view of the above-mentioned shortcomings of the prior art, the present invention aims to provide an efficient dark-field microscope for online observation of cell cultures, which can realize real-time observation.
To achieve the above and other related objects, the present invention provides a dark-field microscope for on-line observation of cell culture, the microscope comprising at least: illuminating element, movable support, support base, objective, photosensitive element, pipe diameter unit, AD converting unit, power module and control system, illuminating element sets up the top at movable support, movable support sets up on supporting the base, the pipe diameter unit connection is in movable support lower part, objective sets up on the pipe diameter unit and corresponds the below at illuminating element, photosensitive element sets up the light source signal that is used for gathering on the objective in the pipe diameter unit, AD converting unit connects photosensitive element, control system connects AD converting unit, power module is for being used for the darkfield microscope power supply of cell culture on-line observation.
Preferably, the lighting element comprises a circular lamp holder and a plurality of lighting lamps arranged on the lamp holder and arranged in a surrounding manner to form a plurality of concentric circles, and the centers of the concentric circles are consistent with the center of the lamp holder.
Preferably, the lamp holder is at a distance of 10-30 cm from the objective lens.
Preferably, the light source of the illuminating lamp and the objective lens form a dark field, so that the requirement of dark field illumination is met.
Preferably, the illuminating lamp is an LED lamp.
Preferably, the support base includes step motor, slide mechanism and base, slide mechanism includes slider and slide rail, preferably, control system connects and controls step motor, step motor connects the slider, but the slider is through slide rail sliding connection on the base and horizontal migration, the movable support sets up on the slider.
Preferably, the pipe diameter unit is connected to the movable support in a sliding manner, a sliding groove is formed in the position, connected to the movable support, of the pipe diameter unit, and the pipe diameter unit can slide up and down along the sliding groove.
Preferably, the control system comprises:
a movable support control unit for controlling the movement of the movable support;
a lighting element control unit for controlling the switching of the lighting element;
the viewing unit is used for viewing the content observed by the objective lens in the current state;
and the storage unit is used for storing the observed contents needing to be saved.
Preferably, the movable stand has a hollow structure, and the movable stand control unit is disposed in the hollow structure.
Preferably, the photosensitive element is a CCD sensor.
As described above, the dark field microscope for cell culture online observation according to the present invention has the following advantageous effects:
the device can realize continuous and repeated observation of the sample without repeated sampling, and the sample is placed between the photosensitive element and the objective lens by only moving the movable bracket through the control system when the sample is observed each time, so that the operation is more convenient and quicker; meanwhile, the damage to the sample during the preparation of the microscopic sample is reduced, so that the detection result is more objective.
Drawings
FIG. 1 is a schematic view showing the overall structure of a dark field microscope according to the present embodiment;
FIG. 2 is a block diagram of the control system;
fig. 3 shows a schematic diagram of the dark field illumination principle.
The lamp comprises a lamp holder 1, a lamp holder 2 and a movable support; 3. objective lens, 4, photosensitive element, 5, support base, 6, pipe diameter unit, 7, control system, 501, stepping motor, 502, slider, 503, base.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the disclosure of the present invention.
Please refer to fig. 1-3. It should be understood that the structures, ratios, sizes, etc. shown in the drawings and attached to the description are only for understanding and reading the disclosure of the present invention, and are not intended to limit the practical conditions of the present invention, so that the present invention has no technical significance, and any modifications of the structures, changes of the ratio relationships, or adjustments of the sizes, should still fall within the scope of the technical contents of the present invention without affecting the efficacy and the achievable purpose of the present invention. In addition, the terms "upper", "lower", "left", "right", "middle" and "one" used in the present specification are for clarity of description, and are not intended to limit the scope of the present invention, and the relative relationship between the terms and the terms is not to be construed as a scope of the present invention.
The invention provides a dark field microscope for cell culture online observation, which comprises but is not limited to: the device comprises an illuminating element, a movable support 2, a supporting base 5, an objective lens 3, a photosensitive element 4, a pipe diameter unit 6, an A/D conversion unit, a control system and a power module. The illumination element sets up at movable support 2 top, movable support 2 sets up on supporting base 5, pipe diameter unit 6 sets up in movable support 2 lower part, objective 3 sets up on pipe diameter unit 6 and corresponds in illumination element's below, photosensitive element 4 sets up and is used for gathering the light source signal on objective 3 in pipe diameter unit 6, A/D conversion unit connects photosensitive element 4, control system connects A/D conversion unit, power module is for being used for the dark field microscope power supply of cell culture on-line observation.
In the embodiment, the photosensitive element adopts a CCD sensor, and the A/D conversion unit transmits an image signal obtained by the CCD sensor to the control system. The control system further comprises a viewing unit provided in the computer for viewing the content observed by the objective lens in the current state, and a storage unit for storing the observed content to be saved.
In the present embodiment, the lighting element is disposed on the top of the movable support 2, and the lighting element includes, but is not limited to, a circular lamp socket 1, and LED lamps arranged on the circular lamp socket 1 and arranged around 5 concentric circles, the distance from the objective lens to the lamp socket is 25 cm, and may be generally set to 10-30 cm, each concentric circle includes ten to twenty LED lamps, and the light source is provided for the whole device. The lamps on the circumference of each concentric circle may be turned on or off simultaneously.
As shown in fig. 3, the lighting element further includes a lens disposed below the LED lamp, so that light emitted from the LED lamp is refracted after passing through the lens, thereby satisfying the requirement of dark field lighting. The included angle between the light emitted by the lighting element and the horizontal shaft of the objective lens 3 is smaller than an angle A, and the angle A is the included angle between the most central light of the lighting element and the horizontal shaft of the objective lens 3.
In this embodiment, the support base 5 includes a stepping motor 501, a sliding mechanism and a base 503, the sliding mechanism includes a slider 502 and a slide rail, the slider 502 is connected to the base 503 through the slide rail, the stepping motor 501 is connected to the control system and is disposed at one end of the base 503, the slider 502 is connected to the stepping motor 501, and the movable support 2 is disposed on the slider 502. Those skilled in the art will appreciate that the step motor 501 can be implemented by using a common step motor 501 in the market, and the step motor 501 can drive the slide block 502 to move so as to change the relative position of the movable support 2 of the whole microscope.
In this embodiment, pipe diameter unit 6 sliding connection is on movable support 2, and pipe diameter unit 6 connects movable support 2 department and is equipped with the spout, and pipe diameter unit 6 can slide from top to bottom along the spout, sets up the through-hole in the relative both sides of 2 on movable support, sets up the screw on two through-holes respectively, when needs adjust rising pipe diameter unit 6, can loosen the screw, removes pipe diameter unit 6, when adjusting to suitable position, screws up the screw. Of course, this adjustment of raising and lowering is only a fine adjustment, with a maximum adjustment range of 5 cm. Other commercially known snap-like means for the securing can of course also be used. Of course, the control system 7 can be used to control the on and off of the LED lamps as needed to meet the needs of observation.
Referring to fig. 2, to further optimize the design, the control system 7 includes, but is not limited to, a lighting element control unit; a movable support control unit, for example, a user may input a specific value to control the stepping motor and thus the moving distance of the movable support 2; the checking unit is mainly used for checking the content which can be observed through the objective lens in the current state by a user; and the storage unit is mainly used for storing the observed contents which are considered to be required to be stored by the user for reviewing.
In another embodiment, the movable support 2 has a hollow structure, and the lighting element control unit is disposed within the hollow structure.
It will be appreciated by those skilled in the art that the control system described above with respect to the control of the illumination elements, the conversion of the image signals acquired by the CCD and the control of the movement of the support base 5 can be implemented using computers, integrated circuit modules, programmable logic devices, other hardware or existing software modules, as is known in the art.
The use of the device has great advantages when observing samples, in particular cells cultured in cell bags or cell factories. When observing the specimen, the movable holder 2 is moved by the control system 7 so that the entire culture vessel is interposed between the illumination element and the objective lens 3, and when the observation is finished, the movable holder 2 is moved by the control system 7 so as to be apart from the culture vessel, and when the observation is required again, the position of the movable holder 2 is moved by the control system.
Therefore, the invention effectively overcomes various defects in the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be made by those skilled in the art without departing from the spirit and scope of the present invention as defined in the appended claims.
Claims (4)
1. A dark field microscope for on-line observation of cell cultures without moving the sample, the microscope comprising at least: the device comprises an illuminating element, a movable support, a supporting base, an objective lens, a photosensitive element, a pipe diameter unit, an A/D conversion unit, a power module and a control system, wherein the illuminating element is arranged at the top of the movable support, the movable support is arranged on the supporting base, the pipe diameter unit is connected to the lower part of the movable support, the objective lens is arranged on the pipe diameter unit and corresponds to the lower part of the illuminating element, the photosensitive element is arranged in the pipe diameter unit and is used for collecting light source signals on the objective lens, the A/D conversion unit is connected with the photosensitive element, the control system is connected with the A/D conversion unit, the power module supplies power for a darkfield microscope for cell culture online observation, the illuminating element comprises a circular lamp holder and illuminating lamps arranged on the lamp holder in a surrounding mode to form a plurality of concentric circles, the circle center of the concentric circles is consistent with the circle center of the lamp holder, the distance from the lamp holder to the objective lens is 10-30 cm, the illuminating lamps are LED lamps, the illuminating element further comprises lenses, and the lenses are arranged below the LED lamps, so that light emitted by the LED lamps is refracted after passing through the lenses, and the requirements of the darkfield of illumination are met; the included angle between the light rays emitted by the lighting element and the horizontal axis of the objective lens is smaller than an angle A, wherein the angle A is the included angle between the light rays at the center of the lighting element and the horizontal axis of the objective lens; the pipe diameter unit is connected to the movable support in a sliding mode, a sliding groove is formed in the position, connected with the movable support, of the pipe diameter unit, the pipe diameter unit can slide up and down along the sliding groove, and the maximum adjusting range is 5 cm;
the support base includes step motor, slide mechanism and base, slide mechanism includes slider and slide rail, control system connects and controls step motor, step motor connects the slider, but the slider just horizontal migration on the base through slide rail sliding connection, movable support sets up on the slider.
2. The dark field microscope for cell culture online observation without moving a sample according to claim 1, characterized in that: the control system includes:
a movable support control unit for controlling the movement of the movable support;
a lighting element control unit for controlling the switching of the lighting element;
the viewing unit is used for viewing the content observed by the objective lens in the current state;
and the storage unit is used for storing the observed contents needing to be saved.
3. The dark field microscope for on-line observation of cell culture without moving a specimen according to claim 2, characterized in that: the movable support has a hollow structure, and the movable support control unit is arranged in the hollow structure.
4. The dark field microscope for cell culture online observation without moving a sample according to claim 1, characterized in that: the photosensitive element is a CCD sensor.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610325563.7A CN105785560B (en) | 2016-05-16 | 2016-05-16 | Dark field microscope for cell culture on-line observation |
| US16/064,414 US10934513B2 (en) | 2015-12-23 | 2016-12-15 | Fully automated continuous cell culture system |
| PCT/CN2016/110035 WO2017107837A1 (en) | 2015-12-23 | 2016-12-15 | Fully automated continuous cell culture system |
| JP2018533724A JP6889723B2 (en) | 2015-12-23 | 2016-12-15 | Cell culture device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610325563.7A CN105785560B (en) | 2016-05-16 | 2016-05-16 | Dark field microscope for cell culture on-line observation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105785560A CN105785560A (en) | 2016-07-20 |
| CN105785560B true CN105785560B (en) | 2022-11-08 |
Family
ID=56378895
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610325563.7A Active CN105785560B (en) | 2015-12-23 | 2016-05-16 | Dark field microscope for cell culture on-line observation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105785560B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10934513B2 (en) | 2015-12-23 | 2021-03-02 | Shanghai GenBase Biotechnology Co., Ltd. | Fully automated continuous cell culture system |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4921978B2 (en) * | 2004-11-09 | 2012-04-25 | 株式会社カネカ | Cell culture device, image processing device, and cell detection system |
| DE102011003603B4 (en) * | 2011-02-03 | 2019-12-24 | Leica Microsystems (Schweiz) Ag | Transmitted light illumination device for a microscope |
| CN103197413B (en) * | 2013-04-15 | 2017-03-15 | 厦门大学 | The optical microscope imaging method of three-dimensional image can be produced |
| CN103207151A (en) * | 2013-04-18 | 2013-07-17 | 上海科源电子科技有限公司 | On-line cell microscopic observer for biological reactor |
| JP6411763B2 (en) * | 2013-07-18 | 2018-10-24 | オリンパス株式会社 | Microscope system and objective lens unit |
| CN204575517U (en) * | 2015-04-29 | 2015-08-19 | 南京英恩特环境技术有限公司 | A kind of micro-imaging dew point hygrometer of dark field illumination |
| CN205750090U (en) * | 2016-05-16 | 2016-11-30 | 上海吉凯基因科技有限公司 | The online dark field microscope observed is cultivated for cell |
-
2016
- 2016-05-16 CN CN201610325563.7A patent/CN105785560B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN105785560A (en) | 2016-07-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103293162B (en) | Lighting system and method used for dark field detection of defect in spherical optical element surface | |
| CN2800304Y (en) | Composite lighting system for mini size on-line high precision vision measure | |
| CN206876951U (en) | A kind of biomicroscope | |
| CN1537225B (en) | Macro illumination device | |
| CN103913830B (en) | Axiomat | |
| CN105785560B (en) | Dark field microscope for cell culture on-line observation | |
| CN203811888U (en) | Box type microscope | |
| CN201812083U (en) | Digital optical zoom stereo microscope for display | |
| TWM427578U (en) | Stereomicroscope | |
| CN102243367A (en) | Pocket microscope | |
| US10775600B2 (en) | Smart media device platform as an inverse microscopic imaging apparatus | |
| CN212965607U (en) | Microscope for laboratory | |
| US20220270279A1 (en) | Sample imaging via two-pass light-field reconstruction | |
| CN201397421Y (en) | Lighting device of stereo microscope | |
| CN204065546U (en) | A kind of Novel microscope | |
| CN2849753Y (en) | Horizontal microscope | |
| CN205750090U (en) | The online dark field microscope observed is cultivated for cell | |
| CN105549193A (en) | Stereoscopic microscope system with 360-degree omnibearing observation function | |
| CN2473630Y (en) | Oil drop tester | |
| CN1474210A (en) | Laser micro function digital microscope device | |
| WO2022111710A1 (en) | Optical path conversion device and optical system | |
| CN205404944U (en) | Stereo microscope system with 360 all -round observation of degree | |
| CN209728317U (en) | A kind of biomicroscope dimming device and biomicroscope | |
| RU207250U1 (en) | PORTABLE MICROSCOPE | |
| CN2775698Y (en) | Low cost stereomicroscope |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20160720 Assignee: HUADAO (SHANGHAI) BIOPHARMA Co.,Ltd. Assignor: SHANGHAI JI KAI GENE TECHNOLOGY Co.,Ltd. Contract record no.: 2017310000066 Denomination of invention: Dark field microscope used in cell culture online observation License type: Exclusive License Record date: 20171207 |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20171226 Address after: Room 101-7, No. 332, Edison Road, China (Shanghai) free trade test area, Shanghai Applicant after: SHANGHAI GENBASE BIOTECHNOLOGY Co.,Ltd. Address before: 201203 room 101-A, No. 328 Edison Road, Zhangjiang hi tech park, Shanghai, Pudong New Area Applicant before: SHANGHAI JI KAI GENE TECHNOLOGY Co.,Ltd. |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |