CN208224044U - Fluorecyte counting criteria piece - Google Patents

Fluorecyte counting criteria piece Download PDF

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Publication number
CN208224044U
CN208224044U CN201820677468.8U CN201820677468U CN208224044U CN 208224044 U CN208224044 U CN 208224044U CN 201820677468 U CN201820677468 U CN 201820677468U CN 208224044 U CN208224044 U CN 208224044U
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China
Prior art keywords
drop coating
fluorecyte
glass slide
framework
coating area
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CN201820677468.8U
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Chinese (zh)
Inventor
沈鹤柏
梅胜兰
相鸿坤
邢国杰
吴炅
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SHANGHAI BHK BIOTECHNOLOGY Co Ltd
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SHANGHAI BHK BIOTECHNOLOGY Co Ltd
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  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The utility model relates to a kind of fluorecyte counting criteria pieces, including glass slide and coverslip, glass slide upper surface is fixed with drop coating area framework, and drop coating is filled with multicolor fluorescence cell sample liquid inside drop coating area framework, and coverslip covers the upper end frame mouth of drop coating area framework.Deviation or excessive occurs after cell sample liquid can be effectively prevent to be squeezed by coverslip for the utility model, and not only standardization level is high for manufactured standard film, increases count results accuracy, and be conducive to the long-term preservation of standard film.

Description

Fluorecyte counting criteria piece
Technical field
The utility model belongs to the technical field of microscopic count glass slide, more particularly to a kind of fluorecyte counting criteria Piece.
Background technique
Glass slide is widely used in biomedical sample production, and histotomy or cell are placed on glass slide, closed the lid Slide is used as observation after mounting.Existing glass slide is when carrying out smear, due to the cell drop coating that do not fix on glass slide Area, drop coating deviate or overflow vulnerable to extruding when the cell on glass slide is in gland slide, generate nonstandardized technique film-making, difference Greatly, phenomena such as easy to fall off, easy spilling.
Circulating tumor cell (circulating tumor cell, CTC) is the immediate cause for being acknowledged as cancer metastasis, Detection CTC is mainly by means such as multicolor fluorescence labels at present.The appearance of automatic fluorescence microscopic analysis system, can efficiently, accurately Detect the CTC in blood in ground.Automatic fluorescence microscopic analysis system is before use, need to calibrate its count accuracy.
Summary of the invention
The technical problem to be solved by the utility model is to provide a kind of fluorecyte counting criteria pieces, can effectively avoid Drop coating liquid is squeezed to form deviation and spill and leakage, guarantees the standardization of film-making.
Technical solution adopted by the utility model to solve its technical problems is providing a kind of fluorecyte counting criteria piece, Including glass slide and coverslip, the glass slide upper surface is fixed with drop coating area framework, and drop coating is filled out inside drop coating area framework Filled with multicolor fluorescence cell sample liquid, the coverslip covers the upper end frame mouth of drop coating area framework.
The glass slide corresponds to the upper surface inside drop coating area framework equipped with hydrophilic coating.
Drop coating area framework is made of hydrophobic material.
The upper surface of the glass slide is equipped with label area.
The label area is frosting.
The label area is located at the side of glass slide, and drop coating area framework is located at the centre in glass slide transparent glass area.
Drop coating area framework is square framework.
Beneficial effect
In the present invention, glass slide upper surface forms the drop coating of multicolor fluorescence cell sample liquid by drop coating area framework Space, and covered by coverslip, deviation or excessive occurs after cell sample liquid can be effectively prevent to be squeezed by coverslip, Not only standardization level is high for manufactured standard film, increases count results accuracy, and be conducive to the long-term preservation of standard film.
Detailed description of the invention
Fig. 1 is the schematic perspective view of the utility model.
Fig. 2 is the positive structure schematic of the utility model.
Specific embodiment
The present invention will be further illustrated below in conjunction with specific embodiments.It should be understood that these embodiments are merely to illustrate this Utility model rather than limitation the scope of the utility model.In addition, it should also be understood that, in the content for having read the utility model instruction Later, those skilled in the art can make various changes or modifications the utility model, and such equivalent forms equally fall within this Shen It please the appended claims limited range.
A kind of fluorecyte counting criteria piece as depicted in figs. 1 and 2, including glass slide 1 and coverslip 2.
Glass slide 1 is with " microscope slide " (JB/T 8230.3-1997) size and material of machinery industry standard Basic (75mm × 25mm × 1.1mm, glass), during 1 upper surface of glass slide is sticked through poly-D-lysine (PLL), PLUS, AdStar One or more coatings form coating so that 1 surface of glass slide have highly stable hydrophilic effect, can effectively adsorb drop Masking liquid prevents from slipping.
The centre of 1 upper surface of glass slide is fixed with a square drop coating area framework 3, is not limited to square framework, drop coating The unilateral width of area's framework 3 is 1mm, and frame thickness is 0.02-0.1mm, and inside is the sample drop coating area of 10mm × 10mm.Drop coating area framework 3 It is made of polytetrafluoroethylene (PTFE) (PTFE), bisphenol A type epoxy resin or carbon nanotube based super hydrophobic material, there is low Poison, surpass The advantages of strong drug resistance, corrosion resistance, drop coating liquid can be effectively prevent excessive.The side of 1 upper surface of glass slide be 19mm × The label area 4 of 25mm white frosting, for pasting bar code or marking pen label.
Framework 3 inside drop coating in drop coating area is filled with multicolor fluorescence cell sample liquid.Cell sample is using the cream to grow fine Gland cancer system SK-BR-3 cell, but not limited to this cell is added complete culture solution and terminates pancreatin digestion, slowly after pancreatin digests Charging and attacking culture solution to cell is single cell suspension, is washed, and the cell after washing is fixed in centrifugation.It is drawn with pipettor a certain amount of The cell fixed washes away extra fixer to centrifuge tube.Select respectively it is different inspire it is blue, green, red fluorescence Dyeing liquor dyes the cells from light after washing, by excess stain liquid washes clean.Above-mentioned cell precipitation piping and druming takes suitable after mixing Drop coating is measured on glass slide, is counted with biology microscope sem observation, mean concentration is recorded, according to this concentration successively by cell precipitation It is diluted to 10 ± 5/5 μ L, 50 ± 10/5 μ L, 100 ± 20/5 μ L.Take above-mentioned cell solution appropriate respectively, drop coating is to originally The drop coating region of utility model glass slide 1, uniformly spreads out, and dark place is placed, and is divided into blank control N, print L, print M, print H, Cell sample concentration is respectively 0,10 ± 5/5 μ L, 50 ± 10,100 ± 20.
Using the mixture and resinene of glycerol buffer as mountant.Glycerol buffer is that glycerol and carbonate are slow The mixture of fliud flushing, by the glycerol buffer of certain volume, drop coating to cell sample area avoids blue-green fluorescent decaying red glimmering Light is deepened, cellular morphology becomes smaller, becomes phenomena such as irregular.3 outside drop coating resinene of drop coating area framework, careful gland coverslip 2, secured mounting thus allows for long-term preservation.
Fluorecyte on glass slide 1 counts it with fluorescence microscope, and aim cell identifies condition are as follows: light field can Observing intact cell form, in fluorescence, same position can observe green fluorescence and blue-fluorescence respectively off field simultaneously.When metering, Repeated n >=6 as effectively count, and the fluorecyte glass slide after metering is fluorecyte counting criteria piece.
Being provided below one uses this fluorecyte counting criteria piece accurate to the analysis of automatic fluorescence microscopic analysis system Property, the specific embodiment calibrated of analytical precision, the analysis performance indicators such as repdocutbility.
1, precision of analysis
Blank control N, print L, print M, print H are respectively tested 10 times, calculate testing mean according to following formula:
X=(X1+X2+X3+ ...+Xi)/n
In formula: Xi-each test value;N-testing time;
Accuracy is calculated according to the following formula:
Accuracy=1- ∣ X-X0 ∣/X0 × 100%
In formula: X0-standard value;X-testing mean;
2, analytical precision
Using the above-mentioned average value X being calculated, the coefficient of variation is calculated according to the following formula:
CV=SD/X × 100%
In formula: X-measurement average value;Xi-i-th test reading;N-pendulous frequency;SD-standard deviation;CV-variation Coefficient.
3, repdocutbility is analyzed
It was an interval every 0.5 hour, retest three times, calculates average value, tests 4 intervals altogether, takes 5 knots Maximum value (minimum value) in fruit;
Instrument repdocutbility error is calculated according to the following formula:
S=∣ Si-S0∣/S0× 100%
In formula: S0--- the reading average value of initial testing;
Si--- the maximum value (minimum value) in test result average value;I=1,2,3,4,5
4, result
It analyzes after tested, it is accurate to print L, print M, the test of print H to can be calculated automatic fluorescence microscopic analysis system Property be respectively 98.96%, 97.96% and 99.17%, test SD value is respectively 1.67,1.00 and 0.5773, and it is accurate to calculate analysis It is respectively 1.72%, 2.00% and 4.77% that degree, which obtains CV value, and repdocutbility error is respectively 0.34%, 1.33% and 2.70%.
5, conclusion
The Quality Control problem of automatic fluorescence microscopic analysis system is successfully solved using this fluorecyte counting criteria piece.

Claims (7)

1. a kind of fluorecyte counting criteria piece, including glass slide (1) and coverslip (2), it is characterised in that: the glass slide (1) upper surface is fixed with drop coating area framework (3), and the internal drop coating of drop coating area framework (3) is filled with multicolor fluorescence cell sample Liquid, the coverslip (2) cover the upper end frame mouth of drop coating area framework (3).
2. a kind of fluorecyte counting criteria piece according to claim 1, it is characterised in that: the glass slide (1) is corresponding The internal upper surface of drop coating area framework (3) is equipped with hydrophilic coating.
3. a kind of fluorecyte counting criteria piece according to claim 1, it is characterised in that: drop coating area framework (3) It is made of hydrophobic material.
4. a kind of fluorecyte counting criteria piece according to claim 1, it is characterised in that: the glass slide (1) it is upper Surface is equipped with label area (4).
5. a kind of fluorecyte counting criteria piece according to claim 4, it is characterised in that: the label area (4) is mill Sand face.
6. a kind of fluorecyte counting criteria piece according to claim 4, it is characterised in that: the label area (4) is located at The side of glass slide (1), drop coating area framework (3) are located at the centre in glass slide (1) transparent glass area.
7. a kind of fluorecyte counting criteria piece according to claim 1, it is characterised in that: drop coating area framework (3) It is square framework.
CN201820677468.8U 2018-05-08 2018-05-08 Fluorecyte counting criteria piece Active CN208224044U (en)

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Application Number Priority Date Filing Date Title
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Application Number Priority Date Filing Date Title
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109884012A (en) * 2019-03-04 2019-06-14 精微视达医疗科技(武汉)有限公司 Fluorescent microsphere testing piece, its production method and production component
CN110673324A (en) * 2019-09-27 2020-01-10 佛山科学技术学院 Glass slide structure suitable for confocal microscope

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109884012A (en) * 2019-03-04 2019-06-14 精微视达医疗科技(武汉)有限公司 Fluorescent microsphere testing piece, its production method and production component
CN109884012B (en) * 2019-03-04 2022-04-08 精微视达医疗科技(武汉)有限公司 Fluorescent microsphere test piece, manufacturing method and manufacturing assembly thereof
CN110673324A (en) * 2019-09-27 2020-01-10 佛山科学技术学院 Glass slide structure suitable for confocal microscope
CN110673324B (en) * 2019-09-27 2022-04-26 佛山科学技术学院 Glass slide structure suitable for confocal microscope

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