CN108707551A - A kind of cell observation device and cell observation method - Google Patents
A kind of cell observation device and cell observation method Download PDFInfo
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- CN108707551A CN108707551A CN201810678273.XA CN201810678273A CN108707551A CN 108707551 A CN108707551 A CN 108707551A CN 201810678273 A CN201810678273 A CN 201810678273A CN 108707551 A CN108707551 A CN 108707551A
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- cell observation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/04—Flat or tray type, drawers
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
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Abstract
The invention belongs to cell observation technical field, a kind of cell observation device and cell observation method are disclosed.The cell observation device includes：Tissue culture plate which is provided with multiple culture holes for containing culture solution；Filling mechanism is used for needing to be added dropwise the culture hole of culture solution to carry out liquid feeding；Micro-imaging mechanism is used for the culture hole after filling mechanism carries out liquid feeding to the culture hole and carries out cell observation.The present invention passes through the culture hole liquid feeding to needing progress cell observation, so that the liquid level of culture solution becomes flat liquid level from concave meniscus, so that when the entire culture hole of light normal illumination, it will not be the phenomenon that bore edges generate black ring, promote observing effect, when in the prior art observing the cell of small-bore tissue culture plate to solve, it is easy to lead to the problem of black ring in bore edges.
Description
Technical field
The present invention relates to cell observation technical field more particularly to a kind of cell observation device and cell observation methods.
Background technology
Tissue culture plate is a kind of labware for cell culture, is formed using special process, cell can be made to reach
To optimal adsorption state, there are multiple culture holes that can cultivate multigroup cell simultaneously.When using tissue culture plate culture cell, experiment
Cell situation is observed usually using phase contrast microscope in room, but uses phase contrast microscope cost higher.
In addition, small-bore tissue culture plate, since aperture is small, the liquid level of culture solution can be because of liquid infiltration and surface tension
Form obvious concave meniscus.When observing cell, due to the presence of the concave meniscus of culture solution, cause close to hole edge
The light of edge can be refracted, and to occur a black ring of circle at bore edges, can only finally observe the cell of hole immediate vicinity, sternly
Weight interference cell observes result.
It would therefore be highly desirable to need to provide a kind of novel cell observation device and cell observation method to solve the above problems.
Invention content
The purpose of the present invention is to provide a kind of cell observation device and cell observation methods, right in the prior art to solve
When the cell of small-bore tissue culture plate is observed, it is easy to lead to the problem of black ring in bore edges.
For this purpose, the present invention uses following technical scheme：
A kind of cell observation device, including：
Tissue culture plate which is provided with multiple culture holes for containing culture solution；
Filling mechanism is used for needing to be added dropwise the culture hole of culture solution to carry out liquid feeding；
Micro-imaging mechanism, be used for when the filling mechanism to the culture hole carry out liquid feeding after the culture hole into
Row cell observation.
Preferably, the tissue culture plate is 96 orifice plates.
Preferably, the amount that the filling mechanism every time carries out the culture hole liquid feeding is 50 μ l.
Preferably, the micro-imaging mechanism within the time no more than eight minutes to be added dropwise culture solution after the training
It supports hole and carries out cell observation.
Preferably, the micro-imaging mechanism includes along showing source of parallel light, microcobjective that light path is set gradually, anti-
Mirror and camera are penetrated, tissue culture plate is between source of parallel light and microcobjective.
Preferably, the multiple of the microcobjective is 4 times.
Preferably, the cell observation device carries out at 20 DEG C~40 DEG C.
A kind of cell observation method, includes the following steps：
Culture solution is added dropwise into the tissue culture plate for be loaded with culture solution；
Cell observation is carried out to the tissue culture plate after dropwise addition culture solution.
Preferably, the amount that culture solution is added dropwise is 50 μ l.
Preferably, carrying out cell sight to the tissue culture plate after dropwise addition culture solution within the time no more than eight minutes
It examines.
Beneficial effects of the present invention：
The present invention passes through the culture hole liquid feeding to needing progress cell observation so that the liquid level of culture solution is become from concave meniscus
Flat liquid level will not promote observing effect so that when the entire culture hole of light normal illumination the phenomenon that bore edges generate black ring,
When in the prior art observing the cell of small-bore tissue culture plate to solve, it is easy to generate black ring in bore edges
Problem.
Description of the drawings
Fig. 1 is the structural schematic diagram of cell observation device provided by the invention.
In figure：
1, source of parallel light；2, tissue culture plate；3, microcobjective；4, speculum；5, camera；6, filling mechanism.
Specific implementation mode
Technical solution to further illustrate the present invention below with reference to the accompanying drawings and specific embodiments.
As shown in Figure 1, it is a kind of cell observation apparatus structure schematic diagram provided by the invention.The cell observation device is thin
Born of the same parents' culture plate 2, filling mechanism 6 and micro-imaging mechanism, wherein：
Tissue culture plate 2 is equipped with multiple culture holes for containing culture solution, and filling mechanism 6 is used for needing that training is added dropwise
The culture hole of nutrient solution carries out liquid feeding, micro-imaging mechanism be used for culture hole after filling mechanism 6 is to culture hole progress liquid feeding into
Row cell observation.The present invention passes through the culture hole liquid feeding to needing progress cell observation so that the liquid level of culture solution is by concave meniscus
Become flat liquid level, so that when the entire culture hole of light normal illumination, observation effect will not be promoted the phenomenon that bore edges generate black ring
Fruit when in the prior art observing the cell of small-bore tissue culture plate to solve, is easy to generate in bore edges black
The problem of ring.
Specifically, tissue culture plate 2 is preferably 96 orifice plates, it is to be understood that can also be 48 orifice plates and 24 orifice plates etc.
Other specification culture plates, when tissue culture plate 2 is 96 orifice plate, a diameter of 9mm of orifice plate, in the range, culture solution liquid level meeting
Because liquid infiltrates and surface tension forms obvious concave meniscus, the influence that black ring generates is the most serious, therefore more suitable
For cell observation device provided by the invention and cell observation method.
Specifically, the amount that filling mechanism 6 every time carries out culture hole liquid feeding is the preferably 40 μ l of μ l~60, specific real at this
It applies in mode, preferably 50 μ l, according to the experiment of a large amount of repeatability of staff, obtains as drawn a conclusion：When liquid volume added is 30 μ l
Concave meniscus can be made to reach flat liquid level substantially when concave meniscus can not be made to reach flat liquid level, and be added to 40 μ l or more.
Specifically, micro-imaging mechanism carries out the culture hole after dropwise addition culture solution within the time no more than eight minutes thin
Born of the same parents observe, and according to the experiment of a large amount of repeatability of staff, obtain as drawn a conclusion：Liquid level can be extensive by about 2 hours after liquid feeding
Complete concave meniscus is arrived again, and is probably initially formed by 8-10 minutes concave meniscus, i.e., image border (i.e. bore edges) start to occur
Shade (i.e. black ring), therefore should ensure that after liquid feeding and shot in 8 minutes.Liquid level is flat liquid level when taking pictures, and avoids recessed liquid
Black ring phenomenon caused by face, the cell situation that can be intuitive to see in entire culture hole are simultaneously automatically snapped.
Specifically, micro-imaging mechanism include along show light path set gradually source of parallel light 1, microcobjective 3, speculum
4 and camera 5, tissue culture plate 2 is between source of parallel light 1 and microcobjective 3.Specifically, the multiple of microcobjective 3 is 2 times
~40 times, preferably 4 times when being observed cell using the micro-imaging mechanism, compared to for phase contrast microscope, are more saved
About cost.
Specifically, cell observation device carries out at 20 DEG C~40 DEG C, such as can carry out at room temperature, practical to thin
Born of the same parents cultivate observation when temperature be 37 DEG C, at this time culture solution volatilization faster, therefore formed the concave meniscus time it is shorter, i.e., cell observation when
Between should be more below 8 minutes time and shot.
A kind of cell observation method, preferred specific embodiment are the above-mentioned cell observation device provided, are specifically included
Following steps：
Culture solution is added dropwise into the tissue culture plate 2 for being loaded with culture solution, to be added dropwise culture solution after tissue culture plate 2 into
Row cell observation.
Since the aperture of small-bore tissue culture plate 2 (such as 96 orifice plates) is small, the liquid level of culture solution can because liquid infiltration and
Surface tension forms obvious concave meniscus.When observing cell, due to the presence of the concave meniscus of culture solution, cause
Light close to bore edges can be refracted, and to occur a black ring of circle at bore edges, can only finally observe hole immediate vicinity
Cell, severe jamming cell observation result.The present invention passes through the culture hole liquid feeding to needing progress cell observation so that culture
The liquid level of liquid becomes flat liquid level from concave meniscus, so that when the entire culture hole of light normal illumination, will not generate black ring in bore edges
The phenomenon that, observing effect is promoted, when being observed in the prior art the cell of small-bore tissue culture plate to solve, is held
Easily black ring is led to the problem of in bore edges.
Obviously, the above embodiment of the present invention is just for the sake of clearly illustrating examples made by the present invention, and it is pair to be not
The restriction of embodiments of the present invention.For the ordinary skill user of fields, may be used also on the basis of the above description
To make other variations or changes in different ways.There is no necessity and possibility to exhaust all the enbodiments.It is all this
All any modification, equivalent and improvement etc., should be included in the claims in the present invention made by within the spirit and principle of invention
Protection domain within.
Claims (10)
1. a kind of cell observation device, which is characterized in that including：
Tissue culture plate (2) which is provided with multiple culture holes for containing culture solution；
Filling mechanism (6) is used for needing to be added dropwise the culture hole of culture solution to carry out liquid feeding；
Micro-imaging mechanism, be used for when the filling mechanism (6) to the culture hole carry out liquid feeding after the culture hole into
Row cell observation.
2. cell observation device according to claim 1, which is characterized in that the tissue culture plate (2) is 96 orifice plates.
3. cell observation device according to claim 1, which is characterized in that the filling mechanism (6) is every time to the training
It is 50 μ l to support hole and carry out the amount of liquid feeding.
4. cell observation device according to claim 1, which is characterized in that the micro-imaging mechanism is no more than eight points
Cell observation is carried out to the culture hole after dropwise addition culture solution in the time of clock.
5. cell observation device according to claim 1, which is characterized in that the micro-imaging mechanism includes along display light
Source of parallel light (1), microcobjective (3), speculum (4) and the camera (5) that road is set gradually, the tissue culture plate (2) are located at
Between the source of parallel light (1) and the microcobjective (3).
6. cell observation device according to claim 5 and cell observation method, which is characterized in that the microcobjective
(3) multiple is 4 times.
7. according to cell observation device according to any one of claims 1 to 6, which is characterized in that the cell observation device
It is carried out at 20 DEG C~40 DEG C.
8. a kind of cell observation method, which is characterized in that include the following steps：
Culture solution is added dropwise to being loaded in the tissue culture plate (2) of culture solution；
Cell observation is carried out to the tissue culture plate (2) after dropwise addition culture solution.
9. cell observation method according to claim 8, which is characterized in that the amount that culture solution is added dropwise is 50 μ l.
10. cell observation method according to claim 8, which is characterized in that drop within the time no more than eight minutes
The tissue culture plate after culture solution (2) is added to carry out cell observation.
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| CN201810678273.XA CN108707551B (en) | 2018-06-27 | 2018-06-27 | Cell observation method |
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| CN201810678273.XA CN108707551B (en) | 2018-06-27 | 2018-06-27 | Cell observation method |
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| CN108707551B CN108707551B (en) | 2022-12-20 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109374376A (en) * | 2018-11-09 | 2019-02-22 | 上海市农业科学院 | A kind of slice preparation method suitable for mushroom lamella Basidium morphologic observation |
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| CN108707551B (en) | 2022-12-20 |
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