CN1924543A - Blood smear leucocyte dye liquor and its preparing process - Google Patents
Blood smear leucocyte dye liquor and its preparing process Download PDFInfo
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- CN1924543A CN1924543A CN 200610124443 CN200610124443A CN1924543A CN 1924543 A CN1924543 A CN 1924543A CN 200610124443 CN200610124443 CN 200610124443 CN 200610124443 A CN200610124443 A CN 200610124443A CN 1924543 A CN1924543 A CN 1924543A
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Abstract
This invention relates to blood smear leucocyte dye liquid, which is composed of amine salt basic ion surface active agent, polyvinyl pyrrolidon or carbowax and regular leucocyte dye liquid with their constitution proportion as 0.0001%-0.04%,0.0125%-2.5% and 30%-75%. This invention can effectively remove the erythrocyte mixture to properly add the blood thickness to make leucocyte number add within each eye scope.
Description
Technical field
The present invention relates to Arneth's count, particularly relate to leucocyte dye liquor of blood film and preparation method thereof.
Background technology
Arneth's count is meant that leucocyte is divided into five classes in the tip blood, i.e. neutrophil leucocyte, eosinophil, basophilic granulocyte, lymphocyte and monocyte.They respectively have its special physiological function, give differential count according to all kinds of leucocyte color characteristics, draw relative ratio, to observe the variation of quantity, form and quality, disease are had the auxiliary diagnosis meaning.
Blood smear leucocyte is dyeed, and then, by artificial microscopic count or the micro-counting of taking pictures, be one of mode of present Arneth's count.
Carrying out artificial microexamination of blood film or micro-taking pictures during white blood cell count(WBC) at present, several problems below main the existence: (1) is because the red blood cell number is leukocytic 500-1000 times, erythrocytic a large amount of existence, make that smear must be very thin, each leucocyte number within the vision significantly reduces when examining under a microscope like this.And in traditional blood film, under the situation about having, RCO can cause leucocyte to be beyond recognition even to cannot see at all leucocyte on leucocyte, causes bigger error for so leukocytic counting.(2) leukocyte count is less within sweep of the eye owing to each, then needs just can carry out leucocyte number statistics according to a lot of field ranges, makes every blood film carry out the chronic of leucocyte number statistics like this.(3) the conventional used dye liquor of leucocyte blood film dyeing has Switzerland's dye liquor, quick dye liquor, Ji's nurse Sa dye liquor, Switzerland-Ji's nurse Sa dye liquor etc., but dyeing time is long, every blood film dyeing time reaches more than one minute at least, that have even nearly more than 15 minutes, be unfavorable for the analysis of blood film in enormous quantities, more be unfavorable for realizing the automated analysis of instrument.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, provide a kind of blood film dyeing time short, can effectively dissolve on the blood film red blood cell and remove red blood cell residual assorted, and make leucocyte dye liquor that leukocytic form do not change and preparation method thereof simultaneously at lysed erythrocyte, make white blood cell count(WBC), the statistics and analysis of blood film can realize reaching fast robotization.
Blood smear leucocyte dye liquor of the present invention, mainly by quaternary ammonium salt cationic surfactant, tween (Tween), or Qu Latong (Triton), or polyvinylpyrrolidone, or non-ionic surfactant, conventional leucocyte dye liquor such as polyglycol are formed.
In particular, blood smear leucocyte dye liquor of the present invention, mainly by quaternary ammonium salt cationic water phase surfactant mixture, tween (Tween), or Qu Latong (Triton), or polyvinylpyrrolidone, or the methanol solution of non-ionic surfactant such as polyglycol, conventional leucocyte dye liquor are formed.
The percent concentration that blood smear leucocyte dye liquor each component of the present invention accounts for total solution is:
The quaternary ammonium salt cationic surfactant is: 0.0001%-0.04%
Above-mentioned non-ionic surfactant is: 0.0125%-2.5%
Conventional leucocyte dye liquor is: 30%-75%
The said quaternary ammonium salt cationic surfactant of the present invention is the best with DTAB, DTAC, cetyl trimethyl ammonium bromide, OTAC, stearyl dimethyl benzyl ammonium chloride.
The said conventional leucocyte dye liquor of the present invention can be Switzerland's dye liquor, quick dye liquor, Ji's nurse Sa dye liquor, Switzerland-existing conventional leucocyte dye liquors such as Ji's nurse Sa dye liquor, is the best with Switzerland-Ji's nurse Sa dye liquor.
The percent concentration of the total solution of quaternary ammonium salt cationic surfactant comprise of the present invention is not a limiting concentration, when the percent concentration of this season amine salt cationic surfactant is lower than concentration range of the present invention, when using dye liquor of the present invention, can can reach effect of the present invention equally by temperature or the prolongation dyeing time that suitably improves dye liquor; When the percent concentration of this season amine salt cationic surfactant was higher than concentration range of the present invention, when using dye liquor of the present invention, temperature that can be by suitably reducing dye liquor or shorten dyeing time can reach effect of the present invention too.
Dye liquor preparation method of the present invention is as follows:
At first, get one or more of a certain amount of quaternary ammonium salt cationic surfactant, be dissolved in the distilled water, heating is stirred, and obtaining concentration range is the A solution of 0.0005%-0.05%.
Get a certain amount of tween again, or Qu Latong, or polyvinylpyrrolidone, or in the non-ionic surfactant such as polyglycol one or more are dissolved in the methyl alcohol, being made into concentration is the B solution of 0.5%-5%.
Then, getting A solution mixes mutually with B solution, drip conventional leucocyte dye liquor again, with AB mixed liquor mixing, the percent concentration that makes each component account for total solution is 0.0001%-0.04% for the quaternary ammonium salts cationic surfactant, tween or Qu Latong or polyvinylpyrrolidone or polyglycol are 0.0125%-2.5%, and conventional leucocyte dye liquor is 30%-75%.Promptly be prepared into leucocyte dye liquor product of the present invention.
The present invention illustrates advantage of the present invention and actual effect by the dyeing experiment contrast of following with conventional leucocyte dye liquor.
Usually, every liter of blood middle leukocytes content often has the following order of magnitude: 1.＜2.0 * 10
9Individual/liter; 2. 2.0-3.0 * 10
9Individual/liter; 3.＞3.0 * 10
9Individual/liter.
Get leucocyte content＜2.0 * 10
9Individual/liter blood 4ul carry out the dyeing of Switzerland-Ji's nurse Sa, dyeing time is 15 minutes, then, drip the damping fluid of Switzerland-Ji's nurse Sa dye liquor, washing, oven dry, examine under a microscope discovery: the leukocytic RCO of often accompanying is within sweep of the eye being arranged, and the leucocyte that causes is beyond recognition, thereby causes white blood cell count(WBC) to be difficult to carry out.In addition, the leucocyte number is few within sweep of the eye owing to each, brings very big trouble for leukocytic counting.As shown in Figure 1.Dye to carry out blood film with a blood with dye liquor of the present invention, dyeing time only needs 11 seconds, after dripping the damping fluid of Switzerland-Ji's nurse Sa dye liquor, washing, oven dry, examine under a microscope discovery: each leucocyte number showed increased within the vision, the leucocyte morphosis is complete, and color also can be distinguished.As shown in Figure 2.
The present invention is to 2.0-3.0 * 10
9Individual/as to rise the contrast of dyeing of the leukocytic blood film of the order of magnitude, obtain as shown in Figure 3, to the result that carries out blood film with a blood and dye as shown in Figure 4 with dye liquor of the present invention with the coloration result of conventional Ji's nurse Sa dye liquor.
It is 5.5 * 10 that the present invention gets leucocyte content
9Individual/blood of rising the order of magnitude carries out blood film dyeing, the coloration result that obtains conventional Switzerland dye liquor as shown in Figure 5, with dye liquor of the present invention as shown in Figure 6 to the result that carries out blood film with a blood and dye.
The invention has the beneficial effects as follows: can shorten the blood film dyeing time greatly, generally, dyeing time only needs about 11 seconds, because red blood cell and residual assorted can effectively the removal, so can suitably increase the thickness of the blood on the smear, thereby each leucocyte number within the vision is increased, and leukocytic form does not change, its clear in structure, the endochylema colouring discrimination is obvious, help artificial microscopic count, the perhaps micro-counting of taking pictures, the computing machine of being more convenient for carries out image recognition, thereby can accelerate the process of automated analysis blood film.
Description of drawings
Fig. 1 gets leucocyte content＜2.0 * 10
9Individual/liter blood, with the picture as a result that Switzerland-Ji's nurse Sa dyeing liquor dyes.
Fig. 2 gets leucocyte content＜2.0 * 10
9Individual/liter blood, with the picture as a result of dye liquor of the present invention dyeing.
Fig. 3 is to 2.0-3.0 * 109/the rise leukocytic blood film of the order of magnitude, the picture as a result that dyes with Ji's nurse Sa dyeing liquor.
Fig. 4 is to 2.0-3.0 * 10
9Individual/as to rise the leukocytic blood film of the order of magnitude, with the picture as a result of dye liquor dyeing of the present invention.
Fig. 5 is to be 5.5 * 10 to leucocyte content
9Individual/as to rise the blood film of the order of magnitude, as to dye with Switzerland's dyeing liquor picture as a result.
Fig. 6 is to be 5.5 * 10 to leucocyte content
9Individual/as to rise the blood film of the order of magnitude, with the picture as a result of dye liquor dyeing of the present invention.
Embodiment
Embodiment 1
Get the 0.1g DTAB, be dissolved in earlier in the 100ml distilled water, heating is stirred, and obtains concentration and is 0.1% DTAB solution, and then use distilled water diluting, obtains concentration and be 0.001% A solution.
The Tween-80 of getting 15ml is dissolved in the methyl alcohol of 500ml, is made into concentration and is 3% B solution.
Get A solution 80ml and mix mutually, and then drip Switzerland-Ji's nurse Sa dye liquor of 75ml,, promptly get product of the present invention with AB mixed liquor mixing with B solution 10ml.
Embodiment 2
Get the 2g cetyl trimethyl ammonium bromide, be dissolved in earlier in the 100ml distilled water, heating is stirred, and obtains concentration and is 2% cetyl trimethyl ammonium bromide solution, and then use distilled water diluting, obtains concentration and be 0.01% A solution.
The TritonX-100 that gets 20ml is dissolved in the methyl alcohol of 500ml, is made into concentration and is 4% B solution.
Get A solution 60ml and mix mutually, and then drip Switzerland-Ji's nurse Sa dye liquor of 80ml,, promptly get product of the present invention with AB mixed liquor mixing with B solution 10ml.
Embodiment 3
Get the 2.5g OTAC, be dissolved in earlier in the 100ml distilled water, heating is stirred, and obtains concentration and is 2.5% OTAC solution, and then use distilled water diluting, obtains concentration and be 0.05% A solution.
The TritonX-80 that gets 10ml is dissolved in the methyl alcohol of 2000ml, is made into concentration and is 0.5% B solution.
Get A solution 100ml and mix mutually, and then drip Switzerland's dye liquor of 100ml,, promptly get product of the present invention with AB mixed liquor mixing with B solution 20ml.
Embodiment 4
Get the 0.2g stearyl dimethyl benzyl ammonium chloride, be dissolved in earlier in the 100ml distilled water, heating is stirred, and obtains concentration and is 0.2% stearyl dimethyl benzyl ammonium chloride solution, and then use distilled water diluting, obtains concentration and be 0.002% A solution.
The polyvinylpyrrolidone of getting 30ml is dissolved in the methyl alcohol of 1000ml, is made into concentration and is 3% B solution.
Get A solution 50ml and mix mutually, and then drip Ji's nurse Sa dye liquor of 70ml,, promptly get product of the present invention with AB mixed liquor mixing with B solution 10ml.
Get 0.2g DTAB, 0.2g OTAC, be dissolved in earlier in the 100ml distilled water, heating is stirred, and obtains concentration and is 0.4% mixed solution, and then use distilled water diluting, obtains concentration and be 0.006% A solution.
The polyvinylpyrrolidone of getting 30ml is dissolved in the methyl alcohol of 800ml, is made into concentration and is 3.75% B solution.
Get A solution 55ml and mix mutually, and then drip the quick dye liquor of 75ml,, promptly get product of the present invention with AB mixed liquor mixing with B solution 10ml.
Embodiment 6
Get 0.1g hexadecyltrimethylammonium chloride, 0.1g stearyl dimethyl benzyl ammonium chloride, be dissolved in earlier in the 100ml distilled water, heating is stirred, and obtains concentration and is 0.2% mixed solution, and then use distilled water diluting, obtains concentration and be 0.002% A solution.
Polyvinylpyrrolidone, the 15ml soil temperature-60 of getting 15ml are dissolved in the methyl alcohol of 750ml, are made into concentration and are 4% B solution.
Get A solution 65ml and mix mutually, and then drip Ji's nurse Sa dye liquor of 85ml,, promptly get product of the present invention with AB mixed liquor mixing with B solution 15ml.
Embodiment 7
Get the 0.4g stearyl dimethyl benzyl ammonium chloride, be dissolved in earlier in the 100ml distilled water, heating is stirred, and obtains concentration and is 0.4% mixed solution, and then use distilled water diluting, obtains concentration and be 0.003% A solution.
Get in the polyvinylpyrrolidone of 15ml, the methyl alcohol that 15mlTritonX-100 is dissolved in 1000ml, be made into concentration and be 3% B solution.
Get A solution 70ml and mix mutually, and then drip the quick dye liquor of 90ml,, promptly get product of the present invention with AB mixed liquor mixing with B solution 15ml.
Embodiment 8
Get the 5g stearyl dimethyl benzyl ammonium chloride, be dissolved in earlier in the 100ml distilled water, heating is stirred, and obtains concentration and is 5% mixed solution, and then use distilled water diluting, obtains concentration and be 0.004% A solution.
Polyvinylpyrrolidone, 10mlTritonX-100, the 5ml polyglycol of getting 5ml are dissolved in the methyl alcohol of 400ml, are made into concentration and are 5% B solution.
Get A solution 75ml and mix mutually, and then drip Switzerland-Ji's nurse Sa dye liquor of 75ml,, promptly get product of the present invention with AB mixed liquor mixing with B solution 20ml.
Claims (6)
1, a kind of blood smear leucocyte dye liquor, it is characterized in that mainly by the quaternary ammonium salt cationic surfactant one or more, in tween or Qu Latong or polyvinylpyrrolidone or the polyglycol one or more, conventional leucocyte dye liquor forms.
2, dye liquor according to claim 1 is characterized in that described quaternary ammonium salt cationic surfactant is one or more in DTAB or DTAC or cetyl trimethyl ammonium bromide or OTAC or the stearyl dimethyl benzyl ammonium chloride.
3, dye liquor according to claim 1 is characterized in that described conventional leucocyte dye liquor is Switzerland's dye liquor or quick dye liquor or Ji's nurse Sa dye liquor or Switzerland-Ji's nurse Sa dye liquor.
4, dye liquor according to claim 3 is characterized in that described conventional leucocyte dye liquor is Switzerland-Ji's nurse Sa dye liquor.
5, according to claim 1,2,3 described dye liquors, it is characterized in that the percent concentration that each component accounts for total solution is 0.0001%-0.04% for the quaternary ammonium salts cationic surfactant, tween or Qu Latong or polyvinylpyrrolidone or polyglycol are 0.0125%-2.5%, and conventional leucocyte dye liquor is 30%-75%.
6, the preparation method of the described blood smear leucocyte dye liquor of a kind of claim 1 is characterized in that:
(1) get one or more of quaternary ammonium salt cationic surfactant, be dissolved in the distilled water, heating is stirred, and obtaining concentration range is the A solution of 0.0005%-0.05%;
(2) get in tween or Qu Latong or polyvinylpyrrolidone or the polyglycol one or more and be dissolved in the methyl alcohol, being made into concentration is the B solution of 0.5%-5%.
(3) getting A solution mixes mutually with B solution, add conventional leucocyte dye liquor again, mixing, the percent concentration that makes each component account for total solution is 0.0001%-0.04% for the quaternary ammonium salts cationic surfactant, tween or Qu Latong or polyvinylpyrrolidone or polyglycol are 0.0125%-2.5%, and conventional leucocyte dye liquor is 30%-75%.
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|---|---|---|---|
| CN 200610124443 CN1924543A (en) | 2006-09-02 | 2006-09-02 | Blood smear leucocyte dye liquor and its preparing process |
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|---|---|---|---|
| CN 200610124443 CN1924543A (en) | 2006-09-02 | 2006-09-02 | Blood smear leucocyte dye liquor and its preparing process |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104698157A (en) * | 2015-02-13 | 2015-06-10 | 中山市创艺生化工程有限公司 | Agent for blood cell analyzer |
| CN104730236A (en) * | 2015-04-16 | 2015-06-24 | 三诺生物传感股份有限公司 | Protein fixing reagent and application thereof |
| CN105300772A (en) * | 2015-09-30 | 2016-02-03 | 成都华西海圻医药科技有限公司 | Wright-Giemsa compound staining solution and preparation method thereof |
| CN106940266A (en) * | 2017-03-21 | 2017-07-11 | 上海美吉医学检验有限公司 | A kind of dyeing enhancing liquid and colouring method dyed for cell surface |
| CN108593392A (en) * | 2018-01-26 | 2018-09-28 | 广州江元医疗科技有限公司 | A kind of vaginal fluid dyeing liquor and preparation method thereof |
| CN108663252A (en) * | 2018-08-14 | 2018-10-16 | 苏州丰泰医疗用品贸易有限公司 | A kind of method of quick carry out Rui Shi Giemsa stainings |
-
2006
- 2006-09-02 CN CN 200610124443 patent/CN1924543A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104698157A (en) * | 2015-02-13 | 2015-06-10 | 中山市创艺生化工程有限公司 | Agent for blood cell analyzer |
| CN104698157B (en) * | 2015-02-13 | 2017-05-17 | 中山市创艺生化工程有限公司 | Agent for blood cell analyzer |
| CN104730236A (en) * | 2015-04-16 | 2015-06-24 | 三诺生物传感股份有限公司 | Protein fixing reagent and application thereof |
| CN105300772A (en) * | 2015-09-30 | 2016-02-03 | 成都华西海圻医药科技有限公司 | Wright-Giemsa compound staining solution and preparation method thereof |
| CN106940266A (en) * | 2017-03-21 | 2017-07-11 | 上海美吉医学检验有限公司 | A kind of dyeing enhancing liquid and colouring method dyed for cell surface |
| CN108593392A (en) * | 2018-01-26 | 2018-09-28 | 广州江元医疗科技有限公司 | A kind of vaginal fluid dyeing liquor and preparation method thereof |
| CN108593392B (en) * | 2018-01-26 | 2020-09-04 | 广州江元医疗科技有限公司 | Vaginal secretion staining solution and preparation method thereof |
| CN108663252A (en) * | 2018-08-14 | 2018-10-16 | 苏州丰泰医疗用品贸易有限公司 | A kind of method of quick carry out Rui Shi Giemsa stainings |
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Application publication date: 20070307 |