Wright-Giemsa dye liquor preparation method and grinding device

Abstract

The invention provides a Wright-Giemsa dye liquor preparation method and a grinding device, and relates to the technical field of biomedicines.The Wright-Giemsa dye liquor preparation method comprises the steps that raw materials are prepared and placed in the grinding device, the raw materials are heated and ground for a first preset duration, and then constant-temperature standing is conducted for a second preset duration; carrying out secondary grinding for a third preset duration; adding methanol, grinding, standing and precipitating for a fourth preset time period, separating out upper-layer dye liquor, repeating for a plurality of times until a preset volume part of methanol is shared, collecting the separated upper-layer dye liquor into a brown bottle, and uniformly mixing, so as to obtain the Wright-Giemsa dye liquor. The method is applied to the grinding device. The raw materials are ground for multiple times, the raw materials are heated during first-time grinding, dissolution and mixing of dye can be accelerated, interpenetration of dye molecules can be better promoted, and therefore the uniformity and the dyeing effect of dye liquor are improved.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a method for preparing Wright-Gymsa stain and a grinding apparatus. Background Technology

[0002] Wright-Gymsza stain is an indispensable composite cell staining agent in pathological diagnosis, microbiological research, and blood cell morphology analysis. The preparation of Wright-Gymsza stain involves grinding Wright's stain powder and Giemsa stain powder. Due to the toxicity of Wright-Gymsza stain, the operation process must simultaneously meet both standardized and safety requirements.

[0003] The traditional method of preparing Wright-Gymsard dye solution involves manual grinding at room temperature using a mortar and pestle. This results in slow dye dissolution and insufficient mixing, leading to poor uniformity of the dye solution and poor dyeing effect. Summary of the Invention

[0004] In view of the above situation, the present invention provides a method for preparing Wright-Gymsard dye solution and a grinding device, which aims to solve the technical problem that the traditional grinding preparation method uses a grinding bowl at room temperature and is purely manual, resulting in low dye dissolution and mixing efficiency, leading to poor uniformity of the dye solution and poor dyeing effect.

[0005] To achieve the above objectives, the present invention provides the following technical solution: In a first aspect, the present invention provides a method for preparing Wright-Gymsa dye solution and a grinding apparatus, comprising: Step S1: Prepare raw materials and place them in a grinding device. Heat and grind the raw materials for a first predetermined time, and then keep them at a constant temperature for a second predetermined time. Step S2, second grinding for the third predetermined time; Step S3: Add methanol, grind, and let stand to precipitate for a predetermined time to separate the upper dye solution; Step S3, repeat step S4 several times until a predetermined volume of methanol is used to obtain Wright-Gymsa staining solution.

[0006] In some embodiments of the present invention, in step S1, the raw material is heated to 37±0.5°C.

[0007] In some embodiments of the present invention, the first predetermined duration is greater than or equal to 4 hours.

[0008] In some embodiments of the present invention, in step S1, the temperature of the constant temperature settling period is 37±0.5℃.

[0009] In some embodiments of the present invention, the second predetermined duration is 12 to 16 hours.

[0010] In some embodiments of the present invention, during steps S1 to S3, when grinding is involved, the dye liquid adhering to the inner wall of the grinding device above the liquid surface is scraped off to grind all the dye liquid in place.

[0011] In some embodiments of the present invention, in steps S1 to S3, a vortex is formed in the liquid inside the grinding device while grinding, thereby achieving the purpose of stirring the liquid.

[0012] In some embodiments of the present invention, 5000 parts by volume of methanol are used in steps S3 to S4.

[0013] In some embodiments of the present invention, in step S1, the raw materials include: 15-17 parts by weight of Wright's dye powder, 3-4 parts by weight of Gymza dye powder, and 33-35 parts by volume of glycerol.

[0014] In a second aspect, the present invention provides a grinding apparatus, comprising: The grinding bowl has a grinding groove on the inside. A grinding rod is disposed in the grinding tank, one end of which contacts the inner wall of the grinding tank and is able to slide circumferentially along the inner wall of the grinding tank; A stirring assembly is disposed inside the grinding tank for stirring the material located inside the grinding tank; Heating components are used to heat the interior of the grinding tank; A wall scraping assembly is disposed inside the grinding tank and is used to scrape off the material adhering to the upper inner wall of the grinding tank.

[0015] The embodiments of the present invention have at least the following advantages or beneficial effects: This invention involves grinding the raw materials multiple times. During the first grinding, the raw materials are heated, which can effectively improve the dissolution rate and mixing degree of the dye, better promote the mutual penetration between dye molecules, and thus significantly improve the uniformity of the dye solution and the dyeing effect.

[0016] Other features and advantages of the present invention will be set forth in the following description. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 This is a schematic flowchart of the preparation method of Wright-Gymsa staining solution in Example 1; Figure 2 This is a schematic diagram of the grinding device in Example 2; Figure 3 This is a schematic diagram of the grinding device in Example 3.

[0019] icon: 1-Grinding bowl body, 11-Grinding trough, 12-Water bath heating cavity, 13-Discharge port, 14-Modible baffle, 2-Grinding rod, 31-Magnetic stirring rod, 32-Magnetic rotating base, 41-Water pipe, 51-Scraper blade, 52-Connecting arm, 6-Grinding cover, 61-Feeding port, 7-Control box, 71-Grinding switch, 72-Speed ​​adjustment switch, 73-Heating switch. Detailed Implementation

[0020] In the following description, only certain exemplary embodiments are briefly described. As those skilled in the art will recognize, the described embodiments can be modified in various ways without departing from the spirit or scope of the embodiments of the invention.

[0021] In the description of the embodiments of the present invention, it should be understood that the terms "upper", "lower", "top", "bottom", "inner", "outer", "circumferential", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing the embodiments of the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the embodiments of the present invention.

[0022] Furthermore, the term "multiple" means two or more, unless otherwise explicitly specified.

[0023] In the embodiments of the present invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in the embodiments of the present invention according to the specific circumstances.

[0024] The embodiments of the present invention will be described in detail below.

[0025] Example 1

[0026] Firstly, see [the following] Figure 1This embodiment provides a method for preparing Wright-Gymsa staining solution, including: Step S1: Prepare the raw materials according to the formula ratio and place them in the grinding device. Heat the raw materials and grind them for a first predetermined time, and then keep them at a constant temperature for a second predetermined time. Step S2, second grinding for the third predetermined time; Step S3: Add methanol (analytical grade), grind, let stand and precipitate for a fourth predetermined time, and separate (absorb) the upper dye solution; Step S4: Repeat step S3 several times until a predetermined volume number (5000 volume parts) of methanol is used to obtain Wright-Gymsa staining solution. Step S5: Collect the Wright-Gymza stain solution in a brown glass bottle and shake well. It can be stored for a long time after preparation.

[0027] Using the above method, the raw materials are ground multiple times. During the first grinding, the raw materials are heated, which can effectively improve the dissolution rate and mixing degree of the dye, better promote the mutual penetration between dye molecules, and thus significantly improve the uniformity of the dye solution and the dyeing effect.

[0028] In step S1, the raw material is heated to 37±0.5℃ and ground for a first predetermined time; the first predetermined time is greater than or equal to 4 hours. The first grinding is carried out at a temperature of 37±0.5℃, which can accelerate the dissolution and mixing of the dye, better promote the mutual penetration between dye molecules, thereby improving the uniformity of the dye solution and the dyeing effect.

[0029] In step S1, the constant temperature for standing is 37±0.5℃; the second predetermined duration is 12 to 16 hours.

[0030] In step S1, the formula ratio of the raw materials includes: 15-17 parts by weight of Wright's dye powder, 3-4 parts by weight of Gymsa dye powder, and 33-35 parts by volume of glycerol.

[0031] In steps S1 to S4, the raw materials used, such as Wright's dye powder, Gymsa dye powder, glycerin, and methanol, can be increased or decreased proportionally according to the preparation ratio.

[0032] In steps S1 to S3, when grinding is involved, the dye liquid adhering to the inner wall of the grinding device above the liquid surface is scraped off to grind all the dye liquid in place.

[0033] In steps S1 to S3, while grinding, a vortex is formed in the liquid inside the grinding device, thereby achieving the purpose of stirring the liquid.

[0034] Secondly, see Figures 1-2This embodiment provides a grinding device that uses the above-mentioned Wright-Gymsa dye preparation method. The grinding device includes a grinding bowl 1, a grinding rod 2, a stirring assembly, a heating assembly, a wall scraping assembly, and a stirring assembly.

[0035] The grinding bowl 1 has a grinding groove 11 on its inner side.

[0036] The grinding rod 2 is placed inside the grinding tank 11. One end of the grinding rod 2 contacts the inner wall of the grinding tank 11 and can slide circumferentially along the inner wall of the grinding tank 11.

[0037] The stirring assembly is installed inside the grinding tank 11 and is used to stir the material located inside the grinding tank 11.

[0038] The heating element is used to heat the inside of the grinding tank 11.

[0039] The wall scraping assembly is installed inside the grinding tank 11 to scrape off the material adhering to the upper inner wall of the grinding tank 11.

[0040] The stirring assembly and grinding rod 2 work together to achieve a better grinding effect. In steps S1 to S3, during grinding, the wall scraping assembly scrapes down the dye solution from the upper inner wall of the grinding tank 11, facilitating thorough grinding of all the dye solution. The heating assembly allows the raw materials to be heated to 37±0.5℃ during grinding in step S1, accelerating the dissolution and mixing of the dye solution and better promoting the interpenetration of dye molecules, thereby improving the uniformity of the dye solution and the dyeing effect. Simultaneously, the heating assembly can create a constant temperature environment of 37±0.5℃ for temperature-controlled settling. The combination of the grinding bowl 1, grinding rod 2, stirring assembly, wall scraping assembly, and heating assembly allows for mechanical replacement of manual grinding operations, significantly reducing labor intensity and harm to personnel. Compared to manual grinding, mechanical grinding can appropriately extend the grinding time, resulting in finer dye particles (average particle diameter d≤1μm observed under a microscope), allowing for thorough grinding and penetration of dye molecules, achieving a better dyeing effect.

[0041] Example 2 This embodiment is a specific implementation of the components of the grinding device in Embodiment 1.

[0042] See Figures 1-2 The stirring assembly includes a magnetic stirring rod 31 and a magnetic rotating base 32. The magnetic stirring rod 31 is disposed inside the grinding tank 11 and located above the magnetic rotating base 32. The magnetic rotating base 32 is disposed at the bottom of the grinding bowl 1 and can rotate under the drive of a matching motor (not shown in the figure). The magnetic rotating base 32 has a built-in permanent magnet, which generates a rotating magnetic field during rotation. This magnetic field magnetically couples with the magnetic stirring rod 31, thereby driving the magnetic stirring rod 31 to rotate synchronously.

[0043] There are multiple grinding rods 2, and their projections on the vertical plane are arranged in a circular pattern. The spacing between adjacent grinding rods 2 is smaller as they approach the bottom of the grinding tank 11. This allows for maximum coverage of the internal area of ​​the grinding tank 11, achieving all-around grinding and a better grinding effect. In this embodiment, six grinding rods 2 are distributed around the magnetic stirring rod 31 and are all connected to the grinding power center. The grinding power center connects the grinding motor (not shown in the figure) and the grinding rods 2. When the grinding motor is working, it drives the multiple grinding rods 2 to rotate through the grinding power center to achieve grinding. In this embodiment, the magnetic stirring rod 31 is rotatably connected to the grinding power center to facilitate the arrangement of the magnetic stirring rod 31.

[0044] The wall scraping assembly includes a connecting arm 52 and an arc-shaped wall scraping blade 51; the wall scraping blade 51 is located above the grinding rod 2 and is in contact with the upper part of the inner wall of the grinding groove 11; the connecting arm 52 is used to connect the grinding power center and the wall scraping blade 51.

[0045] The heating assembly includes a water bath heating cavity 12 disposed within the grinding bowl 1. The water bath heating cavity 12 has a water inlet and a water outlet, and surrounds the outside of the grinding tank 11. By introducing water at a suitable temperature into the water bath heating cavity 12, the material in the grinding tank 11 can be heated. The magnetic rotating base 32 is not located within the water bath heating cavity 12; the two are physically isolated.

[0046] The grinding device also includes a control box 7, which houses a hot water supply unit (not shown in the figure, such as a combination of a water pump, heater, and water tank). The hot water supply unit is connected to the inlet and outlet of the water bath heating cavity 12 via a water pipe 41. The control box 7 has a grinding switch 71, a speed adjustment switch 72, a stirring switch (not shown in the figure), and a heating switch 73 on its exterior. The grinding switch 71 and the speed adjustment switch 72 control the grinding motor to achieve grinding by the grinding rod 2, scraping action by the wall scraper 51, and speed adjustment. The stirring switch rotates the magnetic rotating base 32, thereby driving the magnetic stirring rod 31 to move, achieving stirring. The heating switch 73 controls the hot water supply unit to heat the hot water entering the water bath heating cavity 12.

[0047] The grinding device also includes a grinding cover 6, which is detachably connected to the top of the grinding bowl 1 to create a relatively sealed environment inside the grinding tank 11, reducing the risk of poisoning. The grinding cover 6 has an openable and closable feeding port 61 for easy addition of materials (Wright's dye powder, Gymsard dye powder, glycerin, and methanol). The grinding cover 6 is made of transparent material for easy observation. The grinding power center is rotatably connected to the grinding cover 6.

[0048] The lower part of the grinding bowl 1 is provided with a discharge port 13. A movable baffle 14 is provided inside the discharge port 13. The movable baffle 14 is slidably sealed to the discharge port 13. The inner side of the movable baffle 14 is flush with the inner wall of the grinding tank 11. The movable baffle 14 is used to adjust the opening of the discharge port 13 so as to collect the upper dye liquor (upper Wright-Gymsa dye liquor) after grinding and settling with methanol.

[0049] One method of using the grinding device in this embodiment: 1. Open the grinding cover 6 or the feeding port 61 on the grinding cover 6, and add an appropriate amount of materials (Wright dye powder, Gymsa dye powder and glycerin) according to the formula ratio as needed, and close the grinding cover 6 or the feeding port 61. 2. Turn on the heating switch 73 to gradually increase the temperature of the hot water until the inside of the grinding tank 11 reaches 37±0.5℃; 3. Turn on the grinding switch 71 and the stirring switch. First, use the speed adjustment switch 72 to rotate the grinding rod 2 at a low speed for a few minutes until the material is evenly mixed. Then, use the speed adjustment switch 72 to increase the speed to achieve high-speed grinding. Grind for about 4 to 6 hours, observing the grinding process along the way. The final grinding time depends on the grinding effect (requirement: average particle diameter d ≤ 1μm as observed under a microscope). Stir simultaneously during the grinding process. After achieving the desired grinding effect, turn off the grinding switch 71 and the stirring switch, while keeping the heating switch 73 on. 4. Keep it at a constant temperature (37±0.5℃) and let it stand overnight; 5. Turn off the heating switch 73, turn on the grinding switch 71, adjust the speed adjustment switch 72, grind at high speed for about 30 minutes, and then turn off the grinding switch 71. 6. After adding a certain amount of methanol, adjust the speed control switch 72 and grind at low speed for about 3 to 5 minutes, then turn off the grinding switch 71 (let it stand for about 15 minutes). 7. Open the outlet 13 and move the baffle downwards from high to low according to the liquid level to adjust the opening of the outlet 13 and release the prepared upper dye liquor. Collect the upper dye liquor in a brown glass bottle. When the liquid level is about 2 cm higher than before the addition of methanol (in order to avoid the high concentration of viscous dye liquor at the bottom flowing out), close the outlet 13. 8. Repeat steps 6 and 7 several times until all the methanol in the formula ratio (5000 parts by volume) is used up; 9. Collect all the dye solutions in a brown glass bottle and shake well. The prepared solution can be stored for a long time.

[0050] Example 3 The difference between this embodiment and Embodiment 2 is that: See Figure 3In this embodiment, the heating assembly includes an electric heating film (not shown in the figure) disposed within the grinding bowl 1, a temperature sensor (not shown in the figure) for monitoring the temperature within the grinding tank 11, and a PID temperature control module (not shown in the figure) within the control box 7. The electric heating film is close to the inner wall of the grinding tank 11, and can heat the material within the grinding tank 11. The electric heating film avoids the magnetic rotating base 32 (preferably, the downward projections of the electric heating film and the magnetic rotating base 32 do not overlap), thus avoiding interference with magnetic field coupling. The temperature sensor and the PID temperature control module work together to achieve precise and continuous temperature control. The PID temperature control module adjusts the heating power of the electric heating film based on real-time feedback from the temperature sensor, controlling temperature fluctuations within ±0.5℃ to achieve precise and continuous temperature control.

[0051] The control box 7 is equipped with a grinding switch 71, a speed adjustment switch 72, a stirring switch (not shown in the figure), a heating switch 73, and a temperature display (not shown in the figure). The grinding switch 71 and the speed adjustment switch 72 control the grinding motor to achieve grinding by the grinding rod 2, scraping action by the wall scraper 51, and speed adjustment. The stirring switch rotates the magnetic rotating base 32, which in turn drives the magnetic stirring rod 31 to move, thus achieving stirring. The heating switch 73 is used to control the working status of the heating film, and the temperature display is used to display the temperature data detected by the temperature sensor. Through the settings of the heating switch 73, temperature sensor, and PID temperature control module, the temperature inside the grinding bowl 1 can be easily controlled.

[0052] Finally, it should be noted that the above are merely preferred embodiments of this application and are not intended to limit this application. For those skilled in the art, this application can have various modifications and variations. Without conflict, the embodiments and features described in the embodiments of this application can be arbitrarily combined with each other. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A method of preparing a Wright-Giemsa stain solution, characterized by, include: Step S1: Prepare raw materials and place them in a grinding device. Heat and grind the raw materials for a first predetermined time, and then keep them at a constant temperature for a second predetermined time. Step S2, second grinding for the third predetermined time; Step S3: Add methanol, grind, and let stand to precipitate for a predetermined time to separate the upper dye solution; Step S4: Repeat step S3 several times until a predetermined volume of methanol is used to obtain Wright-Gymsa staining solution.

2. The method of preparing Wright-Giemsa stain solution according to claim 1, wherein, In step S1, the raw material is heated to 37±0.5℃.

3. The method of preparing Wright-Giemsa stain solution according to claim 2, wherein, The first predetermined duration is greater than or equal to 4 hours.

4. The method of preparing Wright-Giemsa stain solution according to claim 1, wherein, In step S1, the temperature of the constant temperature settling period is 37±0.5℃.

5. The method for preparing Wright-Gymsard staining solution according to claim 4, characterized in that, The second scheduled duration is 12 to 16 hours.

6. The method for preparing Wright-Gymsard staining solution according to claim 1, characterized in that, In steps S1 to S3, when grinding is involved, the dye liquid adhering to the inner wall of the grinding device above the liquid surface is scraped off to grind all the dye liquid in place.

7. The method for preparing Wright-Gymsard staining solution according to claim 1, characterized in that, In steps S1 to S3, while grinding, a vortex is formed in the liquid inside the grinding device, thereby achieving the purpose of stirring the liquid.

8. The method for preparing Wright-Gymsard staining solution according to claim 1, characterized in that, In steps S3 to S4, a total of 5000 parts by volume of methanol is used.

9. The method for preparing Wright-Gymsard staining solution according to any one of claims 1 to 8, characterized in that, In step S1, the raw materials include: 15-17 parts by weight of Wright's dye powder, 3-4 parts by weight of Gymsa dye powder, and 33-35 parts by volume of glycerol.

10. A grinding apparatus, characterized in that, include: The grinding bowl has a grinding groove on the inside. A grinding rod is disposed in the grinding tank, one end of which contacts the inner wall of the grinding tank and is able to slide circumferentially along the inner wall of the grinding tank; A stirring assembly is disposed inside the grinding tank for stirring the material located inside the grinding tank; Heating components are used to heat the interior of the grinding tank; A wall scraping assembly is disposed inside the grinding tank and is used to scrape off the material adhering to the upper inner wall of the grinding tank.

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