CN101477024B - Visible component counting pool, its production method and use method - Google Patents
Visible component counting pool, its production method and use method Download PDFInfo
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- CN101477024B CN101477024B CN 200810241789 CN200810241789A CN101477024B CN 101477024 B CN101477024 B CN 101477024B CN 200810241789 CN200810241789 CN 200810241789 CN 200810241789 A CN200810241789 A CN 200810241789A CN 101477024 B CN101477024 B CN 101477024B
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- component
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- solid phase
- support component
- visible component
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- 238000000034 method Methods 0.000 title claims abstract description 17
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 5
- 239000007790 solid phase Substances 0.000 claims abstract description 29
- 230000000007 visual effect Effects 0.000 claims abstract description 19
- 229920001296 polysiloxane Polymers 0.000 claims abstract description 11
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 239000004005 microsphere Substances 0.000 claims description 14
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- 239000011159 matrix material Substances 0.000 claims description 11
- 239000000203 mixture Substances 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 7
- 239000004033 plastic Substances 0.000 claims description 7
- 239000012153 distilled water Substances 0.000 claims description 6
- 238000011010 flushing procedure Methods 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 3
- 238000013019 agitation Methods 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
- 238000011033 desalting Methods 0.000 claims description 3
- 235000019441 ethanol Nutrition 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 239000002184 metal Substances 0.000 claims description 2
- 238000004140 cleaning Methods 0.000 abstract 1
- 238000001035 drying Methods 0.000 abstract 1
- 150000002148 esters Chemical class 0.000 abstract 1
- 208000015181 infectious disease Diseases 0.000 abstract 1
- 230000002458 infectious effect Effects 0.000 abstract 1
- 238000003825 pressing Methods 0.000 abstract 1
- 150000002632 lipids Chemical class 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 208000032420 Latent Infection Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 208000037771 disease arising from reactivation of latent virus Diseases 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 239000007769 metal material Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention relates to a counting cell for physical components, which comprises a bottom solid phase supporting plane (1), and three or more than three supporting components (2) for controlling counting depth are arranged on the bottom solid phase supporting plane. A manufacturing method thereof comprises the following steps: cleaning the supporting components; drying the supporting components; fully and evenly mixing high vacuum silicone ester and the supporting components according to a volume ratio of 2-15:1; and arranging the supporting components on the bottom solid phase supporting plane to form at least three supporting points. A using method comprises the following steps: adding samples (4) to be tested among the supporting components on the bottom solid phase supporting plane of the counting cell; covering the samples by a cover plate (3), and pressing the cover plate to the lowest position; and placing the counting cell below the microscope to count, wherein a computing formula comprises that: the density of the physical components is equal to average physical component number of each visual field divided by a product of the height of the supporting components multiplying the visual field area of the microscope. The counting cell has the advantages of low cost, simple manufacture, convenient use and long valid period, and can reduce threat of infectious samples to health by once use.
Description
Technical field
The present invention relates to the visible component counting assembly, particularly relate to a kind of visible component counting pool, its and preparation method thereof and detect the method for visible component density with this counting chamber.
Background technology
When the laboratory is measured cell, sperm or other tiny visible component density, usually need to use professional counting chamber (plate).Because there is distributional difference in the visible component density to be measured in dissimilar sample, for guaranteeing the reliability of testing result, therefore dissimilar sample need to use corresponding specialized counting chamber, need to use blood countng chamber as detecting red, leucocyte density in blood preparation, measure sperm concentration in semen sample and need to use sperm count pond etc.But the sample of not all type can obtain the commercialization counting chamber that mensuration demand with it is complementary.The sample that cell density is relatively low needs the darker counting degree of depth and the more counting visual field; Otherwise the sample that cell density is relatively high needs the counting chamber of the more shallow counting degree of depth.And commercial professional counting chamber price is relatively high, especially such as the counting chamber of the specific uses such as sperm count pond.In general, these commercial professional counting chambers are Reusabilities, need manually repeatedly to clean the sample that has latent infection in counting chamber, and this certainly will threaten to operator's health.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of simple, degree of depth can be according to the visible component counting pool, its that detects needs and adjust; The present invention also provides its method for making and detects the method for visible component density with this visible component counting pool, its.
For solving the problems of the technologies described above, the invention provides a kind of visible component counting pool, its, it is characterized in that, comprise solid phase support plane, bottom, be interval with to control three or three above support components of the counting degree of depth on solid phase support plane, described bottom.
Cover plate is stamped at described support component top.
Described support component is microballoon.
Described support component is high with viscosity, difficult volatile material adhesion is on solid phase support plane, bottom.
Described support component is four; Described support component is high with viscosity, difficult volatile vacuum silicone matrix material sticks on solid phase support plane, bottom; Described support component is glass or metal or plastic material; Solid phase support plane, described bottom is slide or plastic slide.
Support component height in each counting chamber is identical, and the described support component height degree of depth required according to visible component counting pool, its decided.
The present invention also provides a kind of method of making described visible component counting pool, its, comprises the steps:
Step 1: clean support component;
Step 2: support component is dry;
Step 3: with vacuum silicone matrix material and support component 2～15: 1 abundant mixing by volume;
Step 4: support component is arranged at solid phase support plane, bottom, forms at least 3 strong points.
In described step 1, the method for washing support component is:
A. soak the 50ml microballoon with absolute ethyl alcohol or acetone, be placed in magnetic stirrer, the ethanol that inclines or acetone.So repeated washing is 2 times.With distilled water flushing 5 times.
B. soak microballoon with 1mol/L HCl, be placed in magnetic stirrer 30 minutes, the acid of desalting of inclining.With distilled water flushing 5 times.
C. with deionized water washing 10 times, magnetic agitation is 2 minutes at every turn.
When support component was microballoon, the method that in described step 4, support component is arranged on solid phase support plane, bottom was the point sample method, and concrete steps are as follows:
A. extract the pintle of 10ml syringe, lipid and the mixture of microspheres of described step 3 added wherein, load onto pintle;
B. drain the air in syringe, No. 10 indenting balls are contained on syringe, stand-by;
C. promote to be equipped with the syringe pintle of lipid and mixture of microspheres, with lipid and mixture of microspheres push out the indenting ball point a little, at solid phase support plane, bottom point sample, the strong point that four of each counting chamber points are square and arrange, the scope of four strong point point samples must not surpass the coverage of cover plate.
The present invention also provides a kind of method of using described visible component counting pool, its to detect visible component density, comprises the steps:
A. get the solid phase support plane, bottom with support component, sample to be measured is added on zone between support component;
B. give cover plate at several support component upper covers, and on cover plate downward gentle pressure, until extreme lower position;
C., the counting chamber that sample will be housed is placed in microscopically and counts several visuals field, calculate the number of average each visual field visible component to be measured, and by formula calculate visible component density: the number ÷ (the counting chamber degree of depth * field of microscope area) of the counting volume of the number ÷ counting chamber of visible component density=average every visual field visible component=average every visual field visible component=average every visual field visible component number ÷ (support component height * field of microscope area).
Beneficial effect of the present invention is: (1) controls the distance of solid phase support plane, bottom and top cover plate with several small support components, can detect the needs of using according to reality, realize making very simple to freely the controlling of the counting chamber degree of depth by the support component (being further the microballoon of changing different-diameter in scheme) of changing differing heights.(2) sample to be measured is added between several strong points, thereby counts the degree of depth according to the theory of support component height acquisition counting chamber, and can calculate thus the density of visible component in sample to be measured, very easy to use.(3) material used be easy to obtain and cost very cheap; (4) visible component counting pool, its of made can be disposable operative installations, has greatly reduced the potential threat of infectiousness sample to health.
Further in technical scheme, microballoon is embedded in that viscosity is high, in difficult volatility vacuum silicone matrix material, easily sets up the strong point by the mode of point sample, and be difficult for coming off from solid phase support plane, bottom, the term of validity can reach 3 years at least at ambient temperature.
Description of drawings
Below in conjunction with the drawings and specific embodiments, the present invention is further elaborated.
Fig. 1 adheres to four microballoon strong point schematic diagram on counting chamber base plane of the present invention.
Fig. 2 adds sample schematic diagram to be counted in counting chamber of the present invention pond.
Fig. 3 adds in counting chamber of the present invention pond to add a cover cover plate and the schematic diagram of exerting pressure after counting sample.
Embodiment
See also Fig. 1～shown in Figure 3, visible component counting pool, its of the present invention, by four, solid phase support plane, bottom 1, middle part control the counting degree of depth support component 2---the cover plate 3 at microballoon and top forms, four microballoons form the square zone that holds sample to be detected on solid phase support plane, bottom.Described microballoon is high with viscosity, difficult volatile vacuum silicone matrix material sticks on solid phase support plane, bottom, difficult drop-off.4 microsphere diameters in each counting chamber are identical, so the support component height is identical, form the counting chamber degree of depth of homogeneous.But the different counting chambers that need if the required counting degree of depth is different, can be realized required by the microballoon of choosing different-diameter.
Described support component can be glass or metal material, can be also plastic material; Solid phase support plane, described bottom is slide or plastic slide, can also be the solid phase material of other surfacings.The shape of support component adopts microballoon to make and easy to use, but also can adopt other shapes, such as right cylinder etc.The number of microballoon also can adopt three, the degree of depth that overhangs counting chamber that also can be stable.
The making of visible component counting pool, its of the present invention can be adopted following method:
One, microballoon is prepared
1. soak the 50ml microballoon with absolute ethyl alcohol or acetone, be placed in magnetic stirrer, the ethanol that inclines or acetone.So repeated washing is 2 times.With distilled water flushing 5 times.
2. soak microballoon with 1mol/L HCl, be placed in magnetic stirrer 30 minutes, the acid of desalting of inclining.With distilled water flushing 5 times.
3. with deionized water washing 10 times, magnetic agitation is 2 minutes at every turn.
4. microballoon is toppled over spreading in oven dry in pallet or the inherent baking oven of other Flat bottom containers (note controlling baking temperature when using plastic microsphere and prevent the microballoon distortion), stand-by.
5. pour microballoon vacuum silicone matrix material and above-mentioned processing after into beaker at 2～15: 1 by volume, and abundant mixing, wherein, preferred volume ratio is 5: 1.
6. extract the pintle of 10ml syringe, lipid and mixture of microspheres after step 5 is processed add wherein, load onto pintle.
7. drain the air in syringe, No. 10 indenting balls are contained on syringe, stand-by.
Two, point sample and storage
8. promote to be equipped with the syringe pintle of lipid and mixture of microspheres, with lipid and mixture of microspheres push out the indenting ball point a little, at slide or the surperficial point sample of other smooth solid phase materials, the strong point (referring to Fig. 1) that four of each counting chamber points are square and arrange, the scope of four strong point point samples must not surpass the coverage of cover plate.
9. the slide after point sample vertically is stored in slide box, so that maintain a certain distance between slide, thereby prevents from sticking to the strong point and another slide adhesion on slide.Room temperature preservation, the term of validity can reach 3 years at least.
In conjunction with microscope, use visible component counting pool, its of the present invention to detect the method for visible component density, comprise the steps:
1. in the detection sample during density of visible component, get one of the slide of above-mentioned steps 9 preparations, sample to be measured is added between four strong points that microballoon forms (referring to Fig. 2).
2. cover plate is covered on 4 strong points, and with finger downward gentle pressure on cover plate, until extreme lower position (referring to Fig. 3), namely being assembled into can be for detecting the counting chamber of using.By to the cover plate applying light, make the microballoon that is embedded in matrix material be in sustained height (being the distance of microsphere diameter) in this step.
3. the counting chamber that sample will be housed is placed in microscopically and counts several visuals field, calculate the number of average each visual field visible component to be measured, and by formula calculate visible component density: the number ÷ (the counting chamber degree of depth * field of microscope area) of the counting volume of the number ÷ counting chamber of visible component density=average every visual field visible component=average every visual field visible component=average every visual field visible component number ÷ (microsphere diameter * field of microscope area).
Calculated example: if microsphere diameter 100 μ m, microscope adopts 40 times of object lens and 10 times of wide-angle view eyepieces (aperture is 20mm), and on average the cell number that detects of each visual field is 10, visible component density (/mm
3)=10 ÷ { 0.1 * [(20/40) ÷ 2]
2* π }=509 cell/mm
3
Claims (5)
1. a visible component counting pool, its, is characterized in that, comprises solid phase support plane (1), bottom, is interval with to control three or three above support components (2) of the counting degree of depth on solid phase support plane, described bottom;
Cover plate (3) is stamped at described support component top;
Described support component (2) is microballoon;
Described support component (2) is high with viscosity, difficult volatile vacuum silicone matrix material sticks on solid phase support plane, bottom (1);
Support component height in each counting chamber is identical, and the described support component height degree of depth required according to visible component counting pool, its decided.
2. visible component counting pool, its according to claim 1, is characterized in that, described support component (2) is four; Described support component is glass or metal or plastic material; Solid phase support plane, described bottom is slide or plastic slide.
3. a method of making visible component counting pool, its claimed in claim 1, comprise the steps:
Step 1: clean support component (2);
Step 2: support component is dry;
Step 3: with vacuum silicone matrix material and support component 2～15: 1 abundant mixing by volume;
Step 4: adopt the point sample method that support component is arranged at solid phase support plane (1), bottom, form at least 3 strong points;
In described step 1, the method for washing support component is:
A. soak the 50ml microballoon with absolute ethyl alcohol or acetone, be placed in magnetic stirrer, the ethanol that inclines or acetone; So repeated washing is 2 times; With distilled water flushing 5 times;
B. soak microballoon with 1mol/L HCl, be placed in magnetic stirrer 30 minutes, the acid of desalting of inclining; With distilled water flushing 5 times;
C. with deionized water washing 10 times, magnetic agitation is 2 minutes at every turn.
4. the method for making visible component counting pool, its according to claim 3, is characterized in that, when support component is microballoon, adopts the point sample method that the concrete steps that support component is arranged on solid phase support plane, bottom are as follows in described step 4:
A. extract the pintle of 10ml syringe, vacuum silicone matrix material and the mixture of microspheres of described step 3 added wherein, load onto pintle;
B. drain the air in syringe, No. 10 indenting balls are contained on syringe, stand-by;
C. promote to be equipped with the syringe pintle of vacuum silicone matrix material and mixture of microspheres, with vacuum silicone matrix material and mixture of microspheres push out the indenting ball point a little, at solid phase support plane, bottom point sample, the strong point that four of each counting chamber points are square and arrange, the scope of four strong point point samples must not surpass the coverage of cover plate.
5. right to use requires 1 described visible component counting pool, its to detect the method for visible component density, comprises the steps:
A. get the solid phase support plane, bottom (1) with support component (2), sample to be measured (4) is added on zone between support component;
B. at several support component upper cover cover plates (3), and on cover plate gentle pressure downwards, until extreme lower position;
C., the counting chamber that sample will be housed is placed in microscopically and counts several visuals field, calculate the number of average each visual field visible component to be measured, and by formula calculate visible component density: the number ÷ (the counting chamber degree of depth * field of microscope area) of the counting volume of the number ÷ counting chamber of visible component density=average every visual field visible component=average every visual field visible component=average every visual field visible component number ÷ (support component height * field of microscope area).
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200810241789 CN101477024B (en) | 2008-12-31 | 2008-12-31 | Visible component counting pool, its production method and use method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200810241789 CN101477024B (en) | 2008-12-31 | 2008-12-31 | Visible component counting pool, its production method and use method |
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| Publication Number | Publication Date |
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| CN101477024A CN101477024A (en) | 2009-07-08 |
| CN101477024B true CN101477024B (en) | 2013-05-08 |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103196816B (en) * | 2013-04-08 | 2014-11-12 | 桂林优利特医疗电子有限公司 | Flow type counting chamber with detection inner cavity with adjustable thickness |
| CN105670919A (en) * | 2016-01-05 | 2016-06-15 | 上海北昂精密光电仪器股份有限公司 | Cover-pressing type disposable sperm counting chamber |
| CN106053165A (en) * | 2016-05-16 | 2016-10-26 | 常州大学 | Novel optical method for observing microscopic structure of mucoprotein of sputum |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2297731Y (en) * | 1997-05-28 | 1998-11-18 | 徐元诚 | Counting plate for sperm |
| CN2472233Y (en) * | 2001-03-09 | 2002-01-16 | 彭钧 | Cell counter |
-
2008
- 2008-12-31 CN CN 200810241789 patent/CN101477024B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2297731Y (en) * | 1997-05-28 | 1998-11-18 | 徐元诚 | Counting plate for sperm |
| CN2472233Y (en) * | 2001-03-09 | 2002-01-16 | 彭钧 | Cell counter |
Non-Patent Citations (1)
| Title |
|---|
| JP特开2007-303958A 2007.11.22 |
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| CN101477024A (en) | 2009-07-08 |
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Effective date of registration: 20150211 Address after: 518000, Guangdong, Baoan District, Fuyong Shenzhen street, Huafeng science and Technology Park, tenth blocks C, seventh floor Patentee after: BRED Life Science Technology Inc. Address before: 518049, 93, Meilin Village, Shenzhen, Guangdong, Futian District Patentee before: Liu Yu |
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