CN101477024A - Visible component counting pool, its production method and use method - Google Patents

Abstract

Translated fromChinese

一种有形成份计数池，包括底部固相支撑平面(1)，其上设有用来控制计数深度的三个或三个以上支撑部件(2)。其制作方法包括如下步骤：清洗支撑部件；将支撑部件干燥；将高度真空硅酮酯与支撑部件按体积比2～15∶1充分混匀；将支撑部件设置于底部固相支撑平面，形成至少3个支撑点。使用方法：待测标本(4)加于计数池底部固相支撑平面的支撑部件之间；盖予盖片(3)，将盖片压至最低位置；计数池置于显微镜下计数，计算公式为：有形成份密度＝平均每视野有形成份个数÷(支撑部件高度×显微镜视野面积)。本发明计数深度可据检测需要调整支撑部件高度来获得。成本低，制作简单，使用方便，有效期长，一次性使用可减少传染性标本对健康的威胁。

A formed component counting pool comprises a bottom solid support plane (1), on which three or more support components (2) for controlling the counting depth are arranged. The production method comprises the following steps: cleaning the supporting part; drying the supporting part; fully mixing the high-vacuum silicone ester and the supporting part according to the volume ratio of 2-15:1; setting the supporting part on the bottom solid-phase supporting plane to form at least 3 support points. Method of use: add the sample to be tested (4) between the supporting parts of the solid phase support plane at the bottom of the counting cell; cover with a cover slip (3), press the cover slip to the lowest position; place the counting cell under a microscope for counting, and calculate the formula It is: density of formed components = average number of formed components per field of view ÷ (height of supporting part × field of view of microscope). The counting depth of the present invention can be obtained by adjusting the height of the supporting part according to the detection requirement. The method is low in cost, simple in preparation, convenient in use, long in validity period, and can reduce the health threat of infectious specimens for one-time use.

Description

A kind of visible component counting chamber and preparation method thereof and using method

Technical field

The present invention relates to the visible component counting assembly, particularly relate to a kind of visible component counting chamber and preparation method thereof and use this counting chamber to detect the method for visible component density.

Background technology

When laboratory pair cell, sperm or other tiny visible component density are measured, need to use professional counting chamber (plate) usually.Because there is distributional difference in the visible component density to be measured in the dissimilar samples, for guaranteeing the reliability of testing result, therefore dissimilar samples need use corresponding specialized counting chamber, need use the sperm concentration in blood countng chamber, the mensuration semen sample need use sperm count pond or the like as red, the leucocyte density that detect in the blood preparation.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 higher relatively then needs the counting chamber of the more shallow counting degree of depth.And commercial professional counting chamber price is higher relatively, especially such as the counting chamber of specific uses such as sperm count pond.In general, these commercial professional counting chambers use repeatedly, need manually repeatedly to clean the sample that has latent infection in the 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 provides a kind of simple, degree of depth can be according to detecting the visible component counting chamber that needs are adjusted; The present invention also provides its method for making and uses this visible component counting chamber to detect the method for visible component density.

For solving the problems of the technologies described above, the invention provides a kind of visible component counting chamber, it is characterized in that, comprise solid phase support plane, bottom, on solid phase support plane, described bottom, be interval with three or three above support components being used for controlling the counting degree of depth.

Cover plate is stamped at described support component top.

Described support component is a microballoon.

Described support component with viscosity height, difficult volatile material adhesion on solid phase support plane, bottom.

Described support component is four; Described support component sticks on the solid phase support plane, bottom with viscosity height, difficult volatile vacuum silicone matrix material; 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 described support component height is decided according to the required degree of depth of visible component counting chamber.

The present invention also provides a kind of method of making described visible component counting chamber, comprises the steps:

Step 1: clean support component;

Step 2: with the support component drying;

Step 3: with high vacuum silicone fat and the support component abundant mixing of 2～15:1 by volume;

Step 4: support component is arranged at solid phase support plane, bottom, forms at least 3 strong points.

The method of washing support component is in the described step 1:

A. soak the 50ml microballoon with absolute ethyl alcohol or acetone, place 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, placed magnetic stirrer 30 minutes, the acid of desalting of inclining.With distilled water flushing 5 times.

C. use deionized water wash 10 times, each magnetic agitation 2 minutes.

When support component was microballoon, the method that in the described step 4 support component is arranged on the solid phase support plane, bottom was the point sample method, and concrete steps are as follows:

A. extract the pintle of 10ml syringe, the lipid of described step 3 and mixture of microspheres are added wherein, load onto pintle;

B. drain the air in the syringe, No. 10 indenting balls are contained on the 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 chamber to detect visible component density, comprises the steps:

A. get the solid phase support plane, bottom that has support component, sample to be measured is added on zone between the support component;

B. give cover plate at several support component loam cakes, and on cover plate downward gentle pressure, until extreme lower position;

C., the counting chamber that sample will be housed places microscopically to count 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) is with the several small support component control solid phase support planes, bottom and the distance of top cover plate, can be according to the needs of actual detected application, realize making very simple by the support component (further being the microballoon of changing different-diameter in the scheme) of changing differing heights to freely the controlling of the counting chamber degree of depth.(2) sample to be measured is added between several strong points, thereby obtains the theory counting degree of depth of counting chamber, and can calculate the density of visible component in the sample to be measured thus according to the support component height, very easy to use.(3) used material be easy to obtain and cost very cheap; (4) the visible component counting chamber of made can be disposable operative installations, has significantly reduced the potential threat of infectiousness sample to health.

Further in the technical scheme microballoon is embedded in viscosity height, the 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 synoptic diagram on the counting chamber base plane of the present invention.

Fig. 2 waits to count the sample synoptic diagram for adding in the counting chamber of the present invention pond.

Fig. 3 adds a cover cover plate and the synoptic diagram of exerting pressure after adding waits to count sample in the counting chamber of the present invention pond.

Embodiment

See also Fig. 1～shown in Figure 3, visible component counting chamber of the present invention, by solid phase support plane, bottom 1, middle part four control countings degree of depth support component 2---the cover plate 3 at microballoon and top is formed, four microballoons form the square zone that holds sample to be detected on solid phase support plane, bottom.Described microballoon sticks on the solid phase support plane, bottom with viscosity height, difficult volatile vacuum silicone matrix material, 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 counting chamber of different needs is if required counting degree of depth difference then can realize required by the microballoon of choosing different-diameter.

Described support component can be glass or metal material, also can be 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 chamber of the present invention can be adopted following method:

One, microballoon is prepared

1. soak the 50ml microballoon with absolute ethyl alcohol or acetone, place 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, placed magnetic stirrer 30 minutes, the acid of desalting of inclining.With distilled water flushing 5 times.

3. use deionized water wash 10 times, each magnetic agitation 2 minutes.

4. microballoon is toppled over and sprawl 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.

With the microballoon after high vacuum silicone fat and the above-mentioned processing by volume 2～15:1 pour beaker into, 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 handled add wherein, load onto pintle.

7. drain the air in the syringe, No. 10 indenting balls are contained on the 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 other smooth solid phase material surface point samples, 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 behind the point sample vertically is stored in the slide box, so that maintain a certain distance between slide, thereby prevents to stick to the strong point and another slide adhesion on the slide.Room temperature preservation, the term of validity can reach 3 years at least.

In conjunction with microscope, use visible component counting chamber 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), promptly 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 the matrix material be in sustained height (being the distance of microsphere diameter) in this step.

3. the counting chamber that sample will be housed places microscopically to count 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 detected cell number in each visual field is 10, then visible component density (/mm³)=10 ÷ { 0.1 * [(20/40) ÷ 2]²* π }=509 cell/mm³

Claims (10)

Translated fromChinese

1、一种有形成份计数池，其特征在于，包括底部固相支撑平面(1)，在所述底部固相支撑平面上间隔设有用来控制计数深度的三个或三个以上支撑部件(2)。1. A formed component counting pool is characterized in that it comprises a bottom solid phase support plane (1), and three or more support members (2) for controlling the counting depth are arranged at intervals on the bottom solid phase support plane. ).2、根据权利要求1所述的有形成份计数池，其特征在于，所述支撑部件顶部盖有盖片(3)。2. The formed component counting cell according to claim 1, characterized in that, the top of the support member is covered with a cover sheet (3).3、根据权利要求1或2所述的有形成份计数池，其特征在于，所述支撑部件(2)为微球。3. The formed component counting cell according to claim 1 or 2, characterized in that the support member (2) is a microsphere.4、根据权利要求1或2所述的有形成份计数池，其特征在于，所述支撑部件(2)以粘稠度高、难挥发性的材料粘附在底部固相支撑平面(1)上。4. The formed component counting cell according to claim 1 or 2, characterized in that, the support member (2) is adhered to the bottom solid phase support plane (1) by a material with high viscosity and low volatility .5、根据权利要求1或2所述的有形成份计数池，其特征在于，所述支撑部件(2)为四个；所述支撑部件以粘稠度高、难挥发性的真空硅酮脂质材料粘附在底部固相支撑平面(1)上；所述支撑部件为玻璃或金属或塑料材质；所述底部固相支撑平面为玻片或塑料载片。5. The formed component counting cell according to claim 1 or 2, characterized in that there are four support members (2); the support members are made of vacuum silicone lipid with high viscosity and low volatility The material is adhered on the bottom solid phase support plane (1); the support component is made of glass, metal or plastic; the bottom solid phase support plane is a glass slide or a plastic slide.6、根据权利要求1或2所述的有形成份计数池，其特征在于，每一个计数池内的支撑部件高度相同，所述支撑部件高度根据有形成份计数池所需的深度而定。6. The formed component counting cell according to claim 1 or 2, characterized in that, the height of the support member in each counting cell is the same, and the height of the support member is determined according to the required depth of the formed component counting cell.7、一种制作权利要求1所述的有形成份计数池的方法，包括如下步骤：7. A method for manufacturing the formed component counting cell according to claim 1, comprising the following steps:步骤一：清洗支撑部件(2)；Step 1: cleaning the supporting part (2);步骤二：将支撑部件干燥；Step 2: drying the supporting parts;步骤三：将高度真空硅酮脂与支撑部件按体积比2～15:1充分混匀；Step 3: Fully mix the high-vacuum silicone grease and the supporting parts at a volume ratio of 2 to 15:1;步骤四：将支撑部件设置于底部固相支撑平面(1)，形成至少3个支撑点。Step 4: Set the support member on the bottom solid phase support plane (1) to form at least 3 support points.8、根据权利要求7所述的制作有形成份计数池的方法，其特征在于，所述步骤一中洗涤支撑部件的方法为：8. The method for manufacturing a formed component counting tank according to claim 7, characterized in that the method for washing the supporting parts in the first step is:A.以无水乙醇或丙酮浸泡50ml微球，置于磁力搅拌器搅拌，倾去乙醇或丙酮。如此重复洗涤2次。用蒸馏水冲洗5次。A. Soak 50ml of microspheres in absolute ethanol or acetone, stir on a magnetic stirrer, and pour off the ethanol or acetone. Repeat the washing 2 times. Rinse 5 times with distilled water.B.用1mol/L HCl浸泡微球，置于磁力搅拌器搅拌30分钟，倾去盐酸。用蒸馏水冲洗5次。B. Soak the microspheres with 1mol/L HCl, place on a magnetic stirrer and stir for 30 minutes, then pour off the hydrochloric acid. Rinse 5 times with distilled water.C.用去离子水洗涤10次，每次磁力搅拌2分钟。C. Wash 10 times with deionized water, stirring magnetically for 2 minutes each time.9、根据权利要求7所述的制作有形成份计数池的方法，其特征在于，当支撑部件为微球时，所述步骤四中将支撑部件设置于底部固相支撑平面上的方法为点样法，具体步骤如下：9. The method for manufacturing a formed component counting cell according to claim 7, characterized in that, when the supporting member is a microsphere, the method of setting the supporting member on the bottom solid-phase support plane in the step 4 is spotting method, the specific steps are as follows:A.拔出10ml注射器的针栓，将所述步骤三的脂质与微球混合物加入其中，装上针栓；A. Pull out the plug of the 10ml syringe, add the lipid and microsphere mixture in step 3, and install the plug;B.排尽注射器中的空气，将10号钝针头装在注射器上，待用；B. Exhaust the air in the syringe, install the No. 10 blunt needle on the syringe, and set aside;C.推动装有脂质与微球混合物的注射器针栓，将脂质与微球混合物推压出钝针头尖少许，在底部固相支撑平面点样，每个计数池点四个呈方形排列的支撑点，四个支撑点点样的范围不得超过盖片的覆盖范围。C. Push the plunger of the syringe containing the mixture of lipid and microspheres, push the mixture of lipid and microspheres out of the blunt needle tip a little, spot the sample on the solid support plane at the bottom, and place four points in each counting pool in a square arrangement The support points of the four support points shall not exceed the coverage of the cover slip.10、使用权利要求1或7所述的有形成份计数池检测有形成份密度的方法，包括如下步骤：10. The method for detecting the density of formed components using the formed component counting cell according to claim 1 or 7, comprising the following steps:A.取带有支撑部件(2)的底部固相支撑平面(1)，将待测标本(4)加于支撑部件之间的区域；A. Take the bottom solid phase support plane (1) with support parts (2), and add the sample to be tested (4) to the area between the support parts;B.在几个支撑部件上盖予盖片(3)，并自盖片上向下轻轻施压，直至最低位置；B. Cover the cover sheet (3) on several supporting parts, and gently press down from the cover sheet until it reaches the lowest position;C.将装有标本的计数池置于显微镜下计数若干个视野，计算平均每个视野待测有形成份的个数，并按公式计算有形成份密度：有形成份密度＝平均每视野有形成份的个数÷计数池的计数体积＝平均每视野有形成份的个数÷(计数池深度×显微镜视野面积)＝平均每视野有形成份个数÷(支撑部件高度×显微镜视野面积)。C. Place the counting pool with the specimen under a microscope to count several fields of view, calculate the average number of formed components to be measured in each field of view, and calculate the formed component density according to the formula: formed component density = average formed component per field of view Number ÷ counting volume of the counting pool = average number of formed elements per field of view ÷ (counting pool depth × microscope field of view) = average number of formed elements per field of view ÷ (supporting part height * microscope field of view).

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