CN112840195B

CN112840195B - Sample dyeing method, smear preparation equipment and dyeing liquid combination

Info

Publication number: CN112840195B
Application number: CN201980066960.9A
Authority: CN
Inventors: 李学荣; 余珊
Original assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Current assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Priority date: 2019-06-27
Filing date: 2019-06-27
Publication date: 2024-05-07
Anticipated expiration: 2039-06-27
Also published as: CN112840195A; WO2020258169A1

Abstract

Relates to a sample dyeing method, smear preparation equipment and a dyeing liquid combination. The dyeing liquid combination comprises a first dyeing liquid and a second dyeing liquid, the first dyeing liquid is a mixture of a first biological dye and a second biological dye, the first biological dye can dye acidophilic substances in a sample, and the second biological dye can dye alkalophilic substances in the sample. The second dye liquor is different from the first dye liquor, the second dye liquor is a mixture of a third biological dye and a buffer solution, and the third biological dye can counterstain an alkalophilic substance or an acidophilic substance in a sample, so that the dyeing effect is enhanced. The sample dyeing method comprises the steps of firstly soaking a sample in a first dye liquor for dyeing, then soaking the sample in a second dye liquor for dyeing, wherein in the whole soaking process, the first dye liquor and the second dye liquor cannot fail in a short period, repeated use of the first dye liquor and the second dye liquor is facilitated, and dyeing cost is saved.

Description

Sample dyeing method, smear preparation equipment and dyeing liquid combination

Technical Field

The application relates to the field of medical detection, in particular to a sample dyeing mode.

Background

Sample staining is a very important step in the detection and analysis of samples. Taking a slide as an example, the traditional slide staining method is basically completed by hand, and is low in efficiency. Thus, a number of automated staining instruments are currently available. The automatic dyeing instruments generally adopt a vertical dip dyeing mode, the dyeing process basically imitates manual operation, and usually, a blood membrane is pre-fixed by using pure dye liquor in a small container, and then, a mixed solution of the dye liquor and buffer solution is prepared according to a certain proportion to soak a slide. After a period of time, the mixture was sucked away and discarded, and then repeatedly rinsed with deionized water.

Summary of The Invention

Technical problem

However, the mixed solution of the dye solution and the buffer solution is ineffective within a short time, for example, about ten minutes, so that precipitation is generated, the dyeing effect is not realized, the mixed solution cannot be recycled, and great waste is caused. Although small and medium containers in the existing automatic staining apparatus are usually made into a sheet shape to reduce the volume, the volume is still significantly larger than the manual dosage due to the influence of the area of the glass slide, so that the staining cost is still high.

Solution to the problem

Technical solution

The application mainly provides a novel sample dyeing method, smear preparation equipment and a dyeing liquid combination.

In one embodiment of the present application, there is provided a method for staining a sample, comprising:

The selection step comprises the following steps: selecting a slide on which a sample is smeared;

Dyeing: the method comprises the steps of immersing a sample in a dyeing tank, taking out the sample, and storing a first dye liquor in the dyeing tank, wherein the first dye liquor is a mixture of a first biological dye and a second biological dye, the first biological dye can dye acidophilic substances in the sample, and the second biological dye can dye alkalophilic substances in the sample;

And (3) counterstaining: taking out the sample after soaking in a counterstain, wherein the counterstain stores a second dye solution which is different from the first dye solution, the second dye solution is a mixture of a third biological dye and a buffer solution, and the third biological dye can dye alkalophilic substances or acidophilic substances in the sample;

the method further comprises the steps of: and selecting the next slide smeared with the sample, and repeating the dyeing step to the counterstaining step.

In one embodiment, the method further comprises the step of dyeing: immersing the sample treated by the dyeing step in a dyeing promoting tank, and taking out the sample, wherein a buffer solution is stored in the dyeing promoting tank;

the dyeing promotion step is located after the dyeing step and before the counterstaining step.

In one embodiment, in the step of dyeing, the sample is immersed in the buffer for a period of time ranging from 1 to 5 minutes.

In one embodiment, the method further comprises a step of cleaning before counterstaining: and immersing the sample treated by the dyeing promotion step in a cleaning tank, and taking out the sample, wherein cleaning liquid is stored in the cleaning tank.

In one embodiment, the method further comprises a step of cleaning before counterstaining: washing the sample with a cleaning liquid, the cleaning liquid being stored in a washing tank;

The pre-counterstain washing step is located after the dyeing step and before the counterstain step.

In one embodiment, the sample cleaning time is 1-5 minutes in the pre-counterstain cleaning step.

In one embodiment, the second dye solution further includes a first biological dye, the third biological dye is a basic biological dye, and the volume ratio of the first biological dye is far smaller than the volume ratio of the third biological dye.

In one embodiment, the first biological dye is eosin dye, the second biological dye is methylene blue dye, and the third biological dye is azure dye.

In one embodiment, the first dye solution is a Rui-Giemsa dye solution, the second dye solution is a mixture of Giemsa dye solution and buffer solution, or the second dye solution is a mixture of Liu B dye solution and buffer solution.

In one embodiment, in the second dye liquor, the volume ratio a of the Giemsa dye liquor or the Liu's B dye liquor to the buffer liquor is 1/30.ltoreq.a.ltoreq.1/10.

In one embodiment, in the counterstaining step, the sample soaking time is 5-15 minutes.

In one embodiment, in the staining step, the sample soaking time is 1 to 5 minutes.

In one embodiment, the method further comprises a second cleaning step: and cleaning the sample treated by the counterstaining step.

In one embodiment, the method further comprises:

When the control unit judges that no sample needs to be subjected to the dyeing step or a first dye liquor recovery instruction sent by a user is received, the first dye liquor in the dyeing tank is recovered to a first dye liquor sealing container for storage, and the next use is waited;

When the control unit judges that a sample needs to be subjected to the dyeing step or a first dye liquor inlet instruction sent by a user is received, the first dye liquor stored in the first dye liquor sealing container is sent to the dyeing tank again so as to recycle the first dye liquor.

In one embodiment, the method further comprises:

When the control unit judges that no sample needs to be subjected to the dyeing step or a first dye liquor sealing instruction sent by a user is received, sealing the first dye liquor in the dyeing tank, and waiting for the next use;

and when the control unit judges that the sample needs to be subjected to the dyeing step or receives a first dye liquor opening instruction sent by a user, removing the cover of the dyeing tank so as to recycle the first dye liquor.

In one embodiment, the method for determining that no sample is required for the staining procedure includes: no sample is in the dyeing tank and no sample is waiting to be sent in, and/or the idle time of the dyeing tank exceeds a set value.

In one embodiment, after the control unit determines that the first dye liquor reaches the set discharge requirement or receives a first dye liquor discharge instruction sent by a user, the control unit discharges the first dye liquor.

In one embodiment, the drain requirement includes a ratio of number of dyeings on the sample to volume of the first dye liquor of more than 2 times/ml and/or a usage time of the first dye liquor of more than 2 days.

In one embodiment, the method further comprises:

when the control unit judges that no sample needs to be subjected to the counterstaining step or a second dye liquor recovery instruction sent by a user is received, the second dye liquor in the counterstaining tank is recovered to a second dye liquor recovery container for storage, and the next use is waited;

And after judging that a sample is needed to be subjected to the counterstaining step or a second dye liquor inlet instruction sent by a user is received by the control unit, the second dye liquor stored in the second dye liquor recovery container is sent to the counterstaining groove again so as to recycle the second dye liquor.

In one embodiment, the method further comprises:

When the control unit judges that no sample needs to be subjected to the counterstaining step or a second dye liquor sealing instruction sent by a user is received, sealing the second dye liquor in the counterstaining groove, and waiting for the next use;

And when the control unit judges that the sample needs to be subjected to the counterstaining step or receives a second dye liquor opening instruction sent by a user, removing the cover of the counterstaining groove so as to recycle the second dye liquor.

In one embodiment, the method for determining that no sample is required for the counterstaining step by the control unit includes: and the counterstain tank and the dyeing tank are both free of samples and no samples are to be fed, and/or the idle time of the counterstain tank exceeds a set value.

In one embodiment, the control unit discharges the second dye liquor after judging that the second dye liquor reaches the set discharge requirement or receiving a second dye liquor discharge instruction sent by a user.

In one embodiment, the method further comprises:

When the control unit judges that no sample needs to be subjected to the cleaning step before counterstaining or a cover cleaning liquid instruction sent by a user is received, the cleaning liquid in the cleaning tank is covered, and the cleaning liquid is waited for the next use;

And when the control unit judges that the sample needs to be subjected to the cleaning step before counterstaining or a cleaning liquid opening instruction sent by a user is received, the cover of the cleaning tank is removed so as to recycle the cleaning liquid.

In one embodiment, the method further comprises:

When the control unit judges that no sample needs to be subjected to the dyeing promotion step or a capping dyeing promotion liquid instruction sent by a user is received, capping the buffer liquid in the dyeing promotion tank, and waiting for the next use;

And when the control unit judges that the sample needs to be subjected to the dyeing promotion step or receives an instruction of opening the dyeing promotion liquid sent by a user, removing the cover of the dyeing promotion tank so as to recycle the buffer liquid.

In one embodiment of the present application, there is provided a smear preparation apparatus including a receiving tank, a transfer mechanism, and a control unit, the receiving tank including:

A staining tank for storing a first staining solution, the first staining solution being a mixture of a first biological dye capable of staining acidophilic substances in a sample and a second biological dye capable of staining alkalophilic substances in the sample, the staining tank having a first opening through which a slide on which the sample is smeared can pass;

A counterstain for storing a second dye liquor different from the first dye liquor, the second dye liquor being a mixture of a third biological dye and a buffer liquor, the third biological dye being capable of staining an alkalophilic or alkalophilic substance in the sample, the counterstain having a second opening through which the slide can pass;

The conveying mechanism is used for picking up the slide and driving the slide to move;

The control unit outputs a control signal to control the conveying mechanism to drive the sample to be soaked in the dyeing tank for dyeing, and then soaked in the counterstain tank for counterstaining.

In one embodiment, the dyeing machine further comprises a first dye liquor path control system and a first dye liquor sealing container, wherein the first dye liquor path control system is respectively communicated with the first dye liquor sealing container and the dyeing tank and is used for recycling the corresponding first dye liquor in the dyeing tank to the first dye liquor sealing container and discharging the first dye liquor in the first dye liquor sealing container to the dyeing tank again.

In one embodiment, the first dye liquor path control system comprises a first dye liquor recovery system and a first dye liquor inlet system, wherein the first dye liquor recovery system is respectively communicated with the first dye liquor sealing container and the dyeing tank and is used for recovering the corresponding first dye liquor in the dyeing tank to the first dye liquor sealing container; the first dye liquor inlet system is respectively communicated with the first dye liquor sealing container and the dyeing tank and is used for discharging the first dye liquor in the first dye liquor sealing container into the dyeing tank again.

In one embodiment, the first dye liquor recovery system comprises a first dye liquor recovery pipeline and a first dye liquor recovery pressure source, wherein the first dye liquor recovery pipeline is respectively communicated with the dyeing tank and the first dye liquor sealing container, and the first dye liquor recovery pressure source pumps the first dye liquor in the dyeing tank into the first dye liquor sealing container;

The first dye liquor liquid inlet system comprises a first dye liquor liquid inlet pipeline and a first dye liquor liquid inlet pressure source, the first dye liquor liquid inlet pipeline is respectively communicated with the dyeing tank and the first dye liquor sealing container, and the first dye liquor liquid inlet pressure source discharges the first dye liquor in the first dye liquor sealing container into the dyeing tank.

In one embodiment, the first dye liquor recovery pipeline and the first dye liquor feed pipeline are the same pipeline, and/or the first dye liquor feed pressure source and the first dye liquor recovery pressure source are the same pressure source.

In one embodiment, after the control unit determines that no sample needs to be dyed or receives a first dye liquor recycling instruction sent by a user, the first dye liquor path control system recycles a corresponding first dye liquor in the dyeing tank to a first dye liquor sealing container; when the control unit judges that a sample needs to be dyed or a first dye liquor inlet instruction sent by a user is received, the first dye liquor path control system discharges the first dye liquor in the first dye liquor sealing container into the dyeing tank again.

In one embodiment, the first dye liquor sealed container is a reagent bottle, which is disposed inside the smear preparation apparatus.

In one embodiment, the dyeing machine further comprises a first dye liquor cover, and when the control unit judges that no sample needs to be dyed or receives a first dye liquor sealing instruction sent by a user, the first dye liquor cover shields a corresponding opening on the dyeing tank and reopens the dyeing tank when the dyeing tank needs to be used.

In one embodiment, the method for determining that no sample needs to be dyed by the control unit includes: the staining tank is sample-free and no sample to be stained is fed in, and/or the staining tank idle time exceeds a set value.

In one embodiment, the dyeing machine further comprises a first dye liquor discharge system, wherein the first dye liquor discharge system is communicated with the dyeing tank or the first dye liquor sealing container, and when the control unit judges that the first dye liquor reaches the discharge requirement or receives a first dye liquor discharge instruction sent by a user, the first dye liquor discharge system discharges the first dye liquor in the dyeing tank or the first dye liquor sealing container.

In one embodiment, the emissions requirement comprises: the ratio of the number of dyeings on the sample to the volume of the first dye liquor is more than 2 times/ml and/or the use time of the first dye liquor is more than 2 days.

In one embodiment, the dyeing machine further comprises a second dye liquor path control system and a second dye liquor recovery container, wherein the second dye liquor path control system is communicated with the second dye liquor sealing container and the counterstain, and is used for recovering the second dye liquor corresponding to the counterstain to the second dye liquor recovery container and re-discharging the second dye liquor in the second dye liquor recovery container to the counterstain.

In one embodiment, the second dye liquor path control system comprises a second dye liquor recovery system and a second dye liquor inlet system, wherein the second dye liquor recovery system is communicated with a second dye liquor recovery container and a counterstain, and is used for recovering the corresponding second dye liquor in the counterstain to the second dye liquor recovery container; the second dye liquor inlet system is communicated with the second dye liquor recovery container and the counterstain, and is used for discharging the second dye liquor in the second dye liquor recovery container into the counterstain again.

In one embodiment, the second dye liquor recovery system comprises a second dye liquor recovery pipeline and a second dye liquor recovery pressure source, wherein the second dye liquor recovery pipeline is respectively communicated with the counterstain tank and the second dye liquor recovery container, and the second dye liquor recovery pressure source pumps the second dye liquor in the counterstain tank into the second dye liquor recovery container;

The second dye liquor liquid inlet system comprises a second dye liquor liquid inlet pipeline and a second dye liquor liquid inlet pressure source, the second dye liquor liquid inlet pipeline is respectively communicated with the counterstain and the second dye liquor recovery container, and the second dye liquor liquid inlet pressure source discharges the second dye liquor in the second dye liquor recovery container into the counterstain.

In one embodiment, the second dye liquor recovery pipeline and the second dye liquor feed pipeline are the same pipeline, and/or the second dye liquor feed pressure source and the second dye liquor recovery pressure source are the same pressure source.

In one embodiment, after the control unit determines that no sample needs to perform the counterstaining operation or receives a second dye liquor recovery instruction sent by a user, the second dye liquor path control system recovers a second dye liquor corresponding to the counterstaining tank to a second dye liquor recovery container;

And after the control unit judges that a sample needs to be subjected to the counterstaining work or receives a second dye liquor inlet instruction sent by a user, the second dye liquor path control system discharges the second dye liquor in the second dye liquor recovery container into the counterstaining groove again.

In one embodiment, the dyeing machine further comprises a second dye liquor cover, and when the control unit judges that no sample needs to be subjected to counterstaining work or receives a second dye liquor sealing instruction sent by a user, the second dye liquor cover is controlled to cover the counterstaining groove; and when the control unit judges that the sample needs to be subjected to the counterstaining work or receives a second dye liquor opening instruction sent by a user, controlling the second dye liquor cover to open the counterstaining groove.

In one embodiment, the receiving well further comprises a staining promotion well for storing a buffer, the staining promotion well having a third opening for the slide to pass through; the control unit outputs a control signal to control the conveying mechanism to drive the sample to sequentially move into the dyeing tank for soaking and dyeing, the dyeing promoting tank for dyeing promotion and the counterstain tank for soaking and counterstain.

In one embodiment, the receiving well further comprises a wash well for holding a cleaning liquid, the wash well having a fourth opening for the slide to pass through; after the sample is subjected to dyeing promotion in the dyeing promotion groove and before the sample is subjected to counterstaining in the counterstain groove, the control unit outputs a control signal to control the conveying mechanism to drive the sample to move into the cleaning groove for cleaning.

In one embodiment, the dyeing tank, the dyeing promotion tank, the cleaning tank and the counterstain tank are sequentially arranged in a queue.

In one embodiment, the receiving well further comprises a wash well for holding a cleaning liquid, the wash well having a fourth opening for the slide to pass through; the control unit outputs a control signal to control the conveying mechanism to drive the sample to sequentially move into the dyeing tank for soaking and dyeing, the cleaning tank for cleaning and the counterstain tank for soaking and counterstaining.

In one embodiment, the cleaning device further comprises a cleaning liquid cover, and when the control unit judges that no sample needs to be subjected to cleaning before counterstaining or a cover cleaning liquid instruction sent by a user is received, the cleaning liquid cover is controlled to cover a corresponding opening on the cleaning tank, and the cleaning tank is opened again when the cleaning tank needs to be used.

In one embodiment, the device further comprises an dyeing promoting liquid cover, and when the control unit judges that no sample needs to be subjected to the dyeing promoting work or receives a sealing cover dyeing promoting liquid instruction sent by a user, the control unit controls the dyeing promoting liquid cover to cover a corresponding opening on the dyeing promoting tank and reopens the dyeing promoting tank when the sample needs to be used.

In one embodiment, the device further comprises a mounting seat and a first driving assembly, wherein the accommodating groove is arranged on the mounting seat, the mounting seat is in transmission connection with the first driving assembly, and the accommodating groove on the mounting seat moves under the driving of the first driving assembly.

In one embodiment, the container further comprises an integral cover for shielding the corresponding opening of the container when the container is not in use.

In one embodiment, the device further comprises a mounting seat and a first driving assembly, the accommodating groove is arranged on the mounting seat, any one of the integral outer cover and the mounting seat is in transmission connection with the first driving assembly, and the device is used for enabling the integral outer cover to move relative to the accommodating groove on the mounting seat under the driving of the first driving assembly, so that the corresponding opening on each accommodating groove is shielded.

In one embodiment, the device further comprises a liquid inlet system, wherein a liquid outlet of the liquid inlet system is communicated with the accommodating groove and used for injecting corresponding liquid into the accommodating groove.

In one embodiment, the application provides a dyeing liquid combination for dyeing a sample, which comprises a first dyeing liquid and a second dyeing liquid, wherein the first dyeing liquid is a mixture of a first biological dye and a second biological dye, the first biological dye can dye acidophilic substances in the sample, the second dyeing liquid is a mixture of a third biological dye and a buffer solution, and the third biological dye can dye acidophilic substances or acidophilic substances in the sample.

In one embodiment, the second biological dye and the third biological dye are the same biological dye.

According to the above embodiment, a first dye solution and a second dye solution are provided, the first dye solution is a mixture of a first biological dye capable of dyeing acidophilic substances in a sample and a second biological dye capable of dyeing alkalophilic substances in the sample. The second dye liquor is different from the first dye liquor, the second dye liquor is a mixture of a third biological dye and a buffer solution, and the third biological dye can counterstain an alkalophilic substance or an acidophilic substance in a sample, so that the dyeing effect is enhanced. The sample is soaked in the first dye liquor for dyeing, and then the sample is soaked in the second dye liquor for dyeing, so that the first dye liquor and the second dye liquor cannot lose efficacy in a short period in the whole soaking process, the repeated use of the first dye liquor and the second dye liquor is facilitated, and the dyeing cost is saved.

Advantageous effects of the invention

Brief description of the drawings

Drawings

FIGS. 1 and 2 are schematic views showing the construction of a smear preparation apparatus according to an embodiment of the present application;

FIG. 3 is a schematic diagram of a smear preparation process according to an embodiment of the application;

FIGS. 4-7 are schematic views illustrating steps of several different embodiments of the sample staining method of the present application;

FIG. 8 is a schematic view of a structure of a receiving groove according to an embodiment of the present application;

FIG. 9 is a schematic diagram of a sample staining apparatus according to an embodiment of the present application;

Fig. 10 is a schematic structural diagram of a sample staining apparatus according to another embodiment of the present application.

Inventive examples

Embodiments of the invention

The application will be described in further detail below with reference to the drawings by means of specific embodiments. Wherein like elements in different embodiments are numbered alike in association. In the following embodiments, numerous specific details are set forth in order to provide a better understanding of the present application. However, one skilled in the art will readily recognize that some of the features may be omitted, or replaced by other elements, materials, or methods in different situations. In some instances, related operations of the present application have not been shown or described in the specification in order to avoid obscuring the core portions of the present application, and may be unnecessary to persons skilled in the art from a detailed description of the related operations, which may be presented in the description and general knowledge of one skilled in the art.

Furthermore, the described features, operations, or characteristics of the description may be combined in any suitable manner in various embodiments. Also, various steps or acts in the method descriptions may be interchanged or modified in a manner apparent to those of ordinary skill in the art. Thus, the various orders in the description and drawings are for clarity of description of only certain embodiments, and are not meant to be required orders unless otherwise indicated.

The numbering of the components itself, e.g. "first", "second", etc., is used herein merely to distinguish between the described objects and does not have any sequential or technical meaning. The term "coupled" as used herein includes both direct and indirect coupling (coupling), unless otherwise indicated.

The inventor has conducted intensive analysis and research on the cause of precipitation of the existing dye liquor, and finally found that chemical reaction can occur between different dyes in the existing dye liquor in a buffer environment, so that precipitation can be generated. Taking Rui-Jiemsa dye liquor as an example, since the Rui-Jiemsa dye liquor contains two main dyes of eosin and methylene blue. When the Rui-Jim dye solution and the buffer solution are mixed, in the buffer solution system, the eosin dye presents negative charge, the methylene blue dye presents positive charge, the eosin dye and the methylene blue dye react to generate precipitation, and the process is irreversible, so that the Rui-Jim dye solution and the buffer solution mixed solution cannot be stored for a long time, and the Rui-Jim dye solution and the buffer solution mixed solution are required to be discarded after being used once, so that great waste is caused.

In this regard, the present embodiment provides a sample staining method for staining a sample. The sample includes, but is not limited to, microorganisms, blood, body fluids, bone marrow fluid, and the like. In order to more clearly express the inventive concept, the following will describe blood (slide) and a smear preparation apparatus for preparing the same.

Referring to fig. 1 and 2 in combination, in one embodiment, a smear preparation apparatus is provided for smear preparation of samples of, but not limited to, microorganisms, blood, body fluids, and bone marrow fluids. The smear preparation apparatus includes a sampling mechanism 1 for drawing a sample, a slide loading mechanism 2 for moving a slide to a working line, a dropping mechanism 3 for loading a sample to a slide, a pushing mechanism 4 for screeding a sample on a slide, a drying mechanism (not shown) for drying a blood film on a slide, and a staining mechanism 5 for staining a slide.

Referring to fig. 1 to 3, in one embodiment, when the sampling mechanism 1 performs sample extraction, samples are first mixed, and then a sampling device (for example, a sampling needle 31) in the sampling mechanism 1 is used to suck samples, which may be puncture samples (a sample container has a cover body through which the sampling device passes) or open samples (a sample container is opened, and the sampling device directly sucks samples from an opening portion), according to different sample containers. Blood sample information detection can be performed to obtain information and alignment information, if necessary. In some embodiments, the sample collection device further comprises a micro sample collection mechanism 6, and the micro sample collection mechanism 6 can directly move the test tube placed by the operator to the direction of the sampling device, or the sampling device can also move the test tube placed by the operator to the direction of the test tube placed by the operator. In other implementations, the micro sample injection mechanism 6 may also move the test tube directly toward the drip mechanism 3, or the drip mechanism 3 may also move toward the test tube placed by the operator, and directly draw the blood sample through the drip mechanism 3 (e.g. a drip needle) for sample loading, so that the need for drawing the blood sample through the sampling mechanism 1 is eliminated, and thus micro and preferential sample injection is achieved. When sampling is completed, blood is ready to be dropped onto the slide via the dropping mechanism 3.

Accordingly, the slide loading mechanism 2 extracts a slide and loads the slide to a corresponding position for a blood dropping operation. In some embodiments, as shown in fig. 3, after the slide extraction operation is completed, slide left-right detection and slide cleaning operations may be performed, and then the slide is reloaded to a set position. The loaded slide can print related information and simultaneously carry out operations such as slide positive and negative detection.

With continued reference to fig. 1 to 3, the blood-dripping needle of the sample-dripping mechanism 3 drips the sample onto the slide, and then performs a slide-pushing operation, and the blood is pushed onto the slide into a blood film shape by the slide-pushing mechanism 4. In general, after the completion of the slide pushing operation, the blood film on the slide can be dried to stabilize the form. As shown in fig. 3, in some embodiments, the slide may be driven to flip before the blood film is dried to meet the corresponding needs. In some embodiments, the dried slide may also be subjected to a drying test to determine the effects of blood film drying. In some embodiments, the dried slide may also be subjected to a blood film unfolding test to determine whether the blood film is unfolded and whether the unfolded state is satisfactory.

Continuing with fig. 1-3, after the slide is pushed (blood smear) can be stained (by staining mechanism 5) or output directly (e.g., placed into slide basket 7 for output).

In view of the above smear preparation apparatus or other sample staining related devices, one embodiment of the present application provides a sample staining method.

Referring to fig. 4, 9 and 10, the sample staining method comprises:

Dyeing step S10: the sample is immersed in the dyeing tank 110 and then taken out, and the dyeing tank 110 stores the first dye solution. The first dye solution is a mixture of a first biological dye capable of staining acidophilic substances in the sample and a second biological dye capable of staining alkalophilic substances in the sample.

Counterstaining step S30: the sample is soaked in the counterstain tank 120 and then taken out, and the counterstain tank 120 stores the second dye solution. The second dye solution is different from the first dye solution, and is a mixture of a third biological dye and a buffer solution. The third biological dye is capable of staining the sample for an alkalophilic or acidophilic substance. Wherein the third biological dye may be the same as or different from the first biological dye or the second biological dye.

The staining step S10 may stain the cytoplasm and the nucleus of the sample, respectively, to thereby form a desired effect. In one embodiment, in the staining step S10, the sample soaking time is 1 to 5 minutes (including 1 minute and 5 minutes, and hereinafter, both the two-end values are included in the time-scale). The soaking time can be set to 1-3 minutes in consideration of the comprehensive factors such as the working efficiency and the dyeing effect.

In the dyeing step S10, the first dye solution is a mixture of a first biological dye and a second biological dye, and the first dye solution mainly includes the two dyes, i.e. the first biological dye and the second biological dye, and may further include auxiliary materials which are commonly used in the art, and are conventional in the art, if necessary. Here, the first dye solution is not limited to contain only two components, i.e., the first biological dye and the second biological dye.

As will be appreciated by those skilled in the art, the first biological dye of the present embodiment is capable of staining the acidophiles in the sample, and is capable of binding to the acidophiles in the sample to change color. For example, the eosinophilic substance includes hemoglobin, eosinophilic particles, etc., and the first bio-dye may be eosin dye that changes red color by binding with the eosinophilic substance. Similarly, the second biological dye being capable of staining the basophils in the sample means that the second biological dye is capable of binding to the basophils in the sample to change color. For example, the basophilic substance includes lymphocyte cytoplasm, basophilic particles, etc., and the second biological dye may be a methylene blue dye capable of binding with the basophilic substance to become bluish violet.

The cell nucleus mainly comprises DNA and alkaline protein, the alkaline protein is easy to be combined with eosin to dye red, and meanwhile, a small amount of weak acidic substances are arranged in the cell nucleus to be combined with methylene blue to dye blue, but the blue reaction is weak because of the small content, and the cell nucleus is dyed into mauve. The oxide azurin for methylene blue can also act similarly to methylene blue.

The sample is immersed in a first dye solution, and a carrier containing the sample (a slide will be described below as a blood-bearing slide) is immersed in the first dye solution for a certain period of time. The counterstaining step S30 can dye the cell nucleus of the sample or other acidophilic substances or alkalophilic substances at other positions again, and the addition of the buffer solution is beneficial to improving the dyeing effect of the third dye on the acidophilic substances or alkalophilic substances, so that the subsequent detection and observation are facilitated. In one embodiment, in the counterstaining step S30, the sample soaking time is 5 to 15 minutes. For example, the time period can be controlled to about 10 minutes.

In the counterstaining step S30, the second dye solution is a mixture of the third biological dye and the buffer solution, and the second dye solution mainly comprises the third biological dye and the buffer solution, and may further be added with auxiliary materials which are commonly used in the art, and are conventional means in the art according to need. The second dye solution is not limited to the two components of the third biological dye and the buffer solution. The buffer means a liquid capable of providing an ionized environment, such as a phosphate solution or the like, and may even include an aqueous liquid capable of forming an ionized environment.

The counterstaining step is used for strengthening the dyeing of the basophilic or acidophilic substance, the third biological dye can be selected according to the actual requirement, for example, the biological dye which can dye the basophilic or acidophilic substance can be the same as the first biological dye or the second biological dye, of course, the third biological dye can also be selected to be different from the first biological dye and the second biological dye, for example, the first biological dye is eosin dye, the second biological dye is methylene blue dye, the third biological dye can be azure dye which can be oxidized by methylene blue dye and is used for dyeing the basophilic substance.

The dyeing method provides two types of dye solutions, firstly, a sample is soaked in a first dye solution for dyeing, at the moment, no buffer solution is added into the first dye solution, and chemical reaction between the first biological dye and the second biological dye can not occur, so that precipitation can not occur. After the dyeing step is completed, the sample is soaked in a second dye solution for dyeing, and at the moment, only one acid or alkaline biological dye (third biological dye) is added into the second dye solution for dyeing the alkalophilic substances or the acidophilic substances in the sample. The third biological dye alone cannot form a precipitate in a buffer environment. In the whole dyeing process, the first dye liquor and the second dye liquor can not lose efficacy in a short period, so that the first dye liquor and the second dye liquor can be repeatedly used, and the dyeing cost is saved. After the staining step S10, the carrier of the sample may carry a part of the first bio-dye into the second dye solution, but the part of the first bio-dye and the second bio-dye are very small, and insufficient to form excessive precipitation with the third bio-dye, so that the second dye solution is less affected.

Although the embodiment uses the first dye liquor and the second dye liquor to dye, and increases the number of the dye liquor compared with the existing method, the timeliness is long because the first dye liquor and the second dye liquor can be repeatedly used to soak a plurality of slides. On average, the staining cost per slide is rather greatly reduced.

In one embodiment, the method further comprises the steps of: selecting the next slide smeared with the sample, and repeating the dyeing step to the counterstaining step. This step can be repeated for multiple uses.

Further, referring to fig. 5, 9 and 10, in one embodiment, the dyeing method further includes a dyeing promotion step S21: the sample treated in the dyeing step S10 is soaked in a dyeing promoting tank 130 and then taken out, and the dyeing promoting tank 130 is stored with buffer solution;

The dyeing promotion step S21 is located after the dyeing step S10 and before the counterstaining step S30.

In the dyeing promotion step S21, the buffer solution can further promote the dyeing efficiency of the first biological dye and the second biological dye in the sample to the sample, and the dyeing effect is improved. Meanwhile, the buffer solution plays a role in dyeing promotion and cleaning. Fewer first biological dyes and fewer second biological dyes are attached to the glass slide after the dyeing promoting step S21, which is beneficial to reducing the chemical reaction of the first biological dyes and the second biological dyes in the subsequent counterstaining step S30.

In one embodiment, in the dyeing-promoting step S21, the sample is immersed in the buffer for a period of 1 to 5 minutes, for example, about 2 minutes.

Further, referring to fig. 6, 9 and 10, in one embodiment, the method further includes a pre-counterstain cleaning step S22: the sample processed by the dyeing promotion step S21 is soaked in a cleaning tank 140 and then taken out, and cleaning liquid is stored in the cleaning tank 140;

the pre-counterstain cleaning step S22 is located after the dyeing promotion step S21 and before the counterstain step S30.

In this step, the cleaning liquid may be a buffer solution or a washing liquid, etc., so that the sample is soaked or washed. In the pre-counterstain cleaning step S22, the sample is cleaned by using a buffer solution or a cleaning solution, so that the first biological dye and the second biological dye carried on the glass slide are further reduced, after the step is performed, the glass slide can hardly bring the first biological dye and the second biological dye into the second dye solution in the counterstain step S30, and the first biological dye, the second biological dye and the third biological dye are prevented from generating precipitation by chemical reaction, so that the second dye solution can be used for a longer time, and the dyeing cost is further reduced. The sample may be immersed in the washing liquid or the buffer solution during washing, or may be washed with the washing liquid or the buffer solution.

In the pre-counterstain cleaning step S22, the sample may be cleaned with the buffer solution or the cleaning solution one or more times, depending on the cleaning effect and the requirement. The cleaning liquid can be deionized water or other cleaning liquid commonly used in the field. Of course, the sample may be washed with a buffer solution, and the staining of the sample with the dye may be further promoted.

The washing time may depend on the washing effect and the object to be achieved, and in one embodiment, the sample washing time is 1 to 5 minutes in the washing step S22 before counterstaining.

In another embodiment, as shown in fig. 7, 9 and 10, the dyeing promotion step S21 may not be performed, but the pre-counterstain washing step S22 may be performed after the dyeing step S10 and before the counterstain step S30. At this time, in the pre-counterstain cleaning step S22, the sample is cleaned with the cleaning liquid, and the cleaning liquid is stored in the cleaning tank 140, and the cleaning liquid may be a cleaning liquid or a buffer solution (in this case, the pre-counterstain cleaning step S22 corresponds to the dyeing promotion step S21).

The pre-counterstain cleaning step S22 can clean the first biological dye and the second biological dye carried on the slide, avoid the first biological dye and the second biological dye from being carried into the second dye solution, and generate precipitation with the third biological dye.

Likewise, the cleaning time may depend on the cleaning effect and the intended purpose, and in one embodiment, the sample cleaning time is 1-5 minutes in the pre-counterstain cleaning step S22.

As a more specific example, the third biological dye is a basic biological dye, and the sample may be stained for an alkalophilic substance, such as methylene blue or azure dye. The second dye liquor also comprises a first biological dye, and the volume ratio of the first biological dye is far smaller than that of the third biological dye, so that precipitation caused by chemical reaction of the first biological dye and the third biological dye is avoided.

Specifically, the first biological dye is eosin dye, and the second biological dye is methylene blue dye. Of course, the first and second biological dyes may be replaced with other dyes that are capable of dyeing both basophils and acidophils, respectively.

Further, in order to meet the usage habit of most clinical departments, in one embodiment, the first dye solution is a Rui-Jim Sage dye solution, and the Rui-Jim Sage dye solution (Rui's dye solution) includes eosin dye and methylene blue dye. The second dye solution is formed by mixing Giemsa dye solution and buffer solution, wherein the Giemsa dye solution contains methylene blue dye as a third biological dye (namely, the third biological dye is the same as the second biological dye).

The Rui's dye solution is prepared by mixing basic dye methylene blue (METHYLENE BLUE) and acid dye Eosin (Eosin Y), and is also called Eosin-methylene blue dye. Wherein the colored part of eosin is anion, the colorless part is cation, and the colored part is acid, so eosin is known as acid dye. Methylene blue is typically a chloride salt that is basic, with the colored portion being cationic and the colorless portion being anionic, just as opposed to eosin.

After the Rui's dye solution is placed for a long time, the methylene blue in the dye solution contains azure because of oxidation, and the methylene blue, the azure and the eosin can dye the cell nucleus into purple red, but cannot dye the cytoplasm into blue, and the redundant methylene blue can dye the cytoplasm into blue.

The glass slide dyed by the method is very similar to the traditional Rui-Giemsa dye liquor manual dyeing method, and has high doctor acceptance degree, does not change the habit of a microscopic doctor, and is easy to popularize.

The second dye liquor can also be replaced by a mixture of other dye liquor with methylene blue dye and buffer liquor, such as Liu's B dye liquor.

Further, in order to improve the dyeing effect in the counterstaining step S30, in the second dye liquor, the volume ratio a of the Giemsa dye liquor or the Liu' S B dye liquor to the buffer solution is 1/30.ltoreq.a.ltoreq.1/10.

Preferably, in one embodiment, the volume ratio a of Giemsa or Liu B dye to buffer in the second dye is 1/25.

Further, referring to fig. 4-7, in one embodiment, the method further includes a second cleaning step: and (3) cleaning the sample treated by the counterstaining step S30.

Likewise, the washing of the sample in this step may be performed with a washing liquid or a buffer. The cleaning mode can also use common cleaning means such as soaking or flushing.

By adopting the dyeing method, the use cost of dyeing by a user can be greatly reduced, and the reagent in each step can be repeatedly used, so that the time effectiveness is very long, and the reagent can be stored in an instrument for a long time and the liquid path structure is very simple. In the limit condition, even the liquid path support is not needed, and the user only fills the reagent into the dyeing tank by himself, so that the whole manufacturing cost of the instrument becomes very low, and the instrument is very favorable for popularization in middle-low end customers.

The sample treated by the dyeing method can be continuously subjected to procedures such as drying and outputting, and the procedures can be realized by adopting the prior art, and are not repeated here.

Further, since the present embodiment provides two types of dye solutions, neither the first dye solution nor the second dye solution will fail in a short period of time during the entire dyeing process, and the first dye solution and the second dye solution are repeatedly used.

In one embodiment, please refer to fig. 10, further comprising:

When the control unit judges that no sample needs to be dyed or receives a first dye liquor recovery instruction sent by a user, the control unit recovers the first dye liquor in the dyeing tank 110 to the first dye liquor sealing container 710 for storage and waits for the next use;

When the control unit determines that there is a sample to be dyed or receives a first dye liquor inlet instruction sent by a user, the first dye liquor stored in the first dye liquor sealing container 710 is sent to the dyeing tank 110 again, so as to reuse the first dye liquor.

Since the first dye liquor usually uses methanol of high purity as solvent, it has very strong volatility, and it is volatilized quickly when exposed to air. The steps can recycle, seal and store the first dye liquor when not in use, so as to avoid volatilization and prolong the service life of the first dye liquor.

Of course, this may be accomplished in other ways than recycling to the first dye liquor seal container 710 for sealed storage when not in use.

For example, in one embodiment, the method further comprises:

when the control unit judges that no sample needs to be dyed or receives a first dye liquor sealing instruction sent by a user, the first dye liquor in the dyeing tank 110 is sealed and waits for the next use;

When the control unit determines that the sample needs to be dyed or receives the instruction of opening the first dye liquor sent by the user, the cover of the dyeing tank 110 is removed to reuse the first dye liquor.

The closure may be driven by a corresponding driving arrangement, one of which will be described below.

The above two ways are only one example of sealing and preserving the first dye liquor, and the two ways can be selected and used according to actual structures and requirements. The two ways can not only reduce the volatilization of the first dye liquor, but also avoid that some sundries in the equipment fall into the first dye liquor, and avoid polluting the first dye liquor.

For the timing of recycling or capping, in one embodiment, the manner in which the control unit determines that no sample needs to undergo a staining procedure includes, but is not limited to: no sample is present in the dye vat 110 and no sample is waiting to be fed and/or the idle time of the dye vat 110 exceeds a set point.

In the case of the first dye liquor according to the application, the frequency of replacement is greatly reduced since it can be reused, in the simplest case the first dye liquor can be replaced manually by the user when it is required. Of course, in order to improve the working efficiency and further realize automation, in one embodiment, after the control unit determines that the first dye liquor reaches the set discharge requirement or receives the first dye liquor discharge instruction sent by the user, the control unit discharges the first dye liquor. This discharge can be through a corresponding first liquor discharge system, which will be described further below. The discharged first dye liquor can be collected and treated uniformly.

The discharge requirements of the first dye liquor may be set according to actual requirements, and in one embodiment, the discharge requirements include, but are not limited to: the ratio of the number of times of dyeing the sample to the volume of the first dye liquor is more than 2 times/ml and/or the service time of the first dye liquor is more than 2 days. For example, when 50ml of the first dye solution is stored in the dyeing tank, the first dye solution may be discharged after the sample is subjected to dyeing operation for 100 times or more in the first dye solution. The number of times of dyeing is not absolutely equal to the number of dyed samples, and when the samples complete a complete dyeing action in the first dye liquor to be dyed once, the same sample can be dyed for a plurality of times in the first dye liquor due to different dyeing requirements or dyeing effects. When the first dye liquor reaches the requirements, the first dye liquor can be manually or automatically discharged and replaced by a user.

Further, the second dye liquor, the cleaning liquor, the dyeing promoting liquor and the like can be respectively provided with a recycling system or a sealing means to realize recycling.

In one embodiment, please refer to fig. 10, further comprising:

when the control unit judges that no sample needs to be subjected to the counterstaining step or receives a second dye liquor recovery instruction sent by a user, the second dye liquor in the counterstaining tank 120 is recovered to a second dye liquor recovery container 720 for storage, and the next use is waited;

When the control unit determines that there is a sample to be subjected to the counterstaining step or receives a second dye liquor inlet instruction sent by the user, the second dye liquor stored in the second dye liquor recovery container 720 is sent to the counterstaining tank 120 again, so as to reuse the second dye liquor.

In addition, in one embodiment, the method may also include:

When the control unit judges that no sample needs to be subjected to the counterstaining step or receives a second dye liquor sealing instruction sent by a user, sealing the second dye liquor in the counterstaining groove 120, and waiting for the next use;

When the control unit judges that the sample needs to be subjected to the counterstaining step or receives a second dye liquor opening instruction sent by a user, the cover of the counterstaining groove 120 is removed so as to reuse the second dye liquor.

Through the mode, the second dye liquor can be recycled, volatilization of the second dye liquor is avoided, meanwhile, sundries in the equipment can be prevented from falling into the first dye liquor, pollution of the first dye liquor is avoided, and accordingly the service life of the second dye liquor is prolonged.

In one embodiment, the method for determining that no sample needs to be counterstained by the control unit includes, but is not limited to: neither the counterstain tank 120 nor the staining tank 110 has a sample and no sample is to be fed in, and/or the counterstain tank 120 is idle for more than a set value.

Similarly, the second dye liquor of the present application can be reused, so that the replacement frequency is greatly reduced, and in the simplest way, the second dye liquor can be manually taken and placed by a user when the second dye liquor needs to be replaced. Of course, in order to improve the working efficiency and further realize automation, in one embodiment, after the control unit determines that the second dye liquor reaches the set discharge requirement or receives the second dye liquor discharge instruction sent by the user, the second dye liquor is discharged.

The discharge requirement may be determined with reference to the discharge requirement of the first dye liquor, for example from the ratio of the number of dyeings on the sample to the volume of the second dye liquor and/or the number of days of use of the second dye liquor. Of course, other emissions requirements are also possible.

Also, referring to fig. 10, for a cleaning liquid and a buffer liquid as the cleaning liquid, in one embodiment, further includes:

When the control unit judges that no sample needs to be subjected to the cleaning step before counterstaining or a cover cleaning liquid instruction sent by a user is received, the cleaning liquid in the cleaning tank 140 is covered and waits for the next use;

When the control unit determines that the sample needs to be subjected to the pre-counterstaining cleaning step or receives a cleaning liquid opening instruction sent by a user, the control unit releases the cover of the cleaning tank 140 so as to reuse the cleaning liquid.

For use as a buffer in the dyeing promoting step, in one embodiment, the method further comprises:

When the control unit judges that no sample needs to be subjected to the dyeing promotion step or receives a cover dyeing promotion liquid instruction sent by a user, the buffer liquid in the cover dyeing promotion tank 130 is covered and waits for the next use;

when the control unit determines that the sample needs to be subjected to the dyeing promotion step or receives an instruction of opening the dyeing promotion liquid sent by a user, the control unit releases the cover of the dyeing promotion tank 130 so as to recycle the buffer liquid.

Since the cleaning solution and the buffer solution are less volatile than the first and second dye solutions, the above recovery of the cleaning solution and the buffer solution may be selected as appropriate. In addition, the discharge and replacement of the cleaning and buffer solutions can also be carried out manually by the user or automatically by means of the device, with reference to the treatment methods of the first and second solutions described above.

Referring to fig. 8 to 10, an embodiment of the present application further provides a sample staining apparatus for a smear preparation apparatus, which includes a receiving tank 100, a transfer mechanism 200, and a control unit (not shown). The sample staining apparatus may be used as a sample staining solution module for smear preparation equipment or other related equipment.

The receiving tank 100 includes a dyeing tank 110 for storing a first dye solution and a counterstain tank 120 for storing a second dye solution.

The first dye liquor may be as defined hereinabove. The staining cell 110 has a first opening through which the carrier with the sample can pass. The second dye liquor may also be a second dye liquor as defined above. The counterstain tank 120 has a second opening through which the carrier can pass.

The transfer mechanism 200 is used to pick up the slide 500 with the sample and to move the slide 500. The control unit outputs a control signal to control the transfer mechanism to drive the sample to be soaked in the dyeing tank 110 for dyeing (the dyeing step S10 is completed), and then soaked in the counterstain tank 120 for counterstaining (the counterstaining step S30 is completed).

The conveying mechanism 200 may use various power sources, such as a motor, a cylinder, a hydraulic cylinder, an electromagnetic drive, etc., to drive the slide 500 to move. In one embodiment, referring to fig. 9 and 10, the transfer mechanism 200 includes a pick-up assembly 210 for picking up a slide 500 and a first drive assembly 220 (the pick-up assembly 210 and the first drive assembly 220 are shown in fig. 9 as simple schematic), the first drive assembly 220 being in driving connection with the pick-up assembly 210 to move the pick-up assembly 210.

The first driving unit 220 may be driven by a motor, a cylinder, a hydraulic cylinder, an electromagnetic drive, or the like, as described above. The picking assembly 210 can then employ various types of robotic structures that can pick and release the slide 500. The first driving component 220 drives the pick-up component to move, for example, as shown in fig. 9, the first driving component 220 drives the pick-up component 210 to move along the y-axis and the z-axis, so as to drive the slide 500 to change positions in the two directions, so as to move to the positions of the receiving slots 100 for soaking. Alternatively, as shown in FIG. 10, the first drive assembly 220 drives the pick-up assembly 210 in three directions, the x-axis, the y-axis, and the z-axis.

Further, referring to fig. 9 and 10, in one embodiment, the receiving chamber 100 further includes a staining promoting chamber 130 for storing a buffer solution, and the staining promoting chamber 130 has a third opening for passing a slide 500. After the sample is dyed in the dyeing tank 110 and before the sample is dyed in the counterstain tank 120, the control unit outputs a control signal to control the conveying mechanism to drive the sample to move into the dyeing promoting tank 130 for dyeing and cleaning (the dyeing promoting step S21 is completed).

Further, referring to fig. 9 and 10, in one embodiment, the accommodating groove 100 further includes a cleaning groove 140 for storing a cleaning solution or a buffer solution, and the cleaning groove 140 has a fourth opening for passing the slide 500; after the sample is finished in the dyeing promoting tank 130 and before the sample is finished in the counterstaining tank 120, the control unit outputs a control signal to control the conveying mechanism 200 to drive the sample to move into the cleaning tank 140 for cleaning (the step S22 of cleaning before counterstaining is finished).

In order to improve the convenience of the operation and to improve the dyeing efficiency, as shown in fig. 9 and 10, in one embodiment, the dyeing tank 110, the dyeing promoting tank 130, the washing tank 140, and the counterstain tank 120 are sequentially arranged in a line, for example, in the y-axis. The picking assembly 210 may sequentially perform corresponding operations through the respective receiving slots 100, and sequentially complete the respective steps. This arrangement can make the sample staining apparatus more compact, and can shorten the movement stroke of the pickup assembly 210, improving the staining efficiency. It is understood that the dyeing tank 110, the dyeing promoting tank 130, the rinsing tank 140, and the counterstain tank 120 may be integrally formed or separately provided.

In yet another embodiment, the dye-enhancing bath 130 may not be present. The receiving groove 100 further includes a washing groove 140 for storing a washing liquid, the washing groove 140 having a fourth opening through which the slide 500 passes; after the sample is dyed in the dyeing tank 110 and before the sample is counterstained in the counterstaining tank 120, the control unit outputs a control signal to control the conveying mechanism 200 to drive the sample to move into the washing tank 140 for washing (the step S22 of washing before counterstaining is completed). The rinse tank 140 may be disposed directly between the dye tank 110 and the counterstain tank 120.

In the above embodiments, the cleaning tank 140 may be configured to perform one or more cleaning operations on the sample with the buffer solution or the cleaning solution, depending on the cleaning effect and the requirement.

Further, the receiving groove 100 further includes a second washing groove 150 for storing a washing liquid or buffer, and the second washing groove 150 has a fifth opening for passing the slide 500. After the sample is counterstained in the counterstaining tank 120, the control unit outputs a control signal to control the conveying mechanism 200 to drive the sample to move into the cleaning tank 140 for cleaning (the second cleaning step S50 is completed).

Further, referring to fig. 8, in one embodiment, some or all of the accommodating tanks 100 (including the dyeing tank 110, the counterstaining tank 120, the dyeing promoting tank 130, the cleaning tank 140 and the second cleaning tank 150) may have the structure shown in fig. 8. The accommodating groove 100 stores a corresponding liquid (e.g., a first dye solution, a second dye solution, a buffer solution, a cleaning solution, etc.) therein and can impregnate the slide 500 coated with the sample. The receiving groove 100 contains at least one first receiving part 101, the first receiving part 101 forming an insertion area 102 for inserting a slide. The corresponding (e.g., staining) process is performed by immersing the slide 500 mounted on the first mounting part 101 in a liquid (e.g., a dye solution) for a certain time in the receiving groove 100.

Further, referring to fig. 9 and 10, in one embodiment, the dyeing tank 110, the counterstaining tank 120, the dyeing promoting tank 130, the rinsing tank 140, and the second rinsing tank 150 are sequentially arranged. The pick-up assembly 210 is more than one. For example, as shown in fig. 10, the number of pickup assemblies 210 is two, so that the pickup assemblies can work simultaneously to improve the working efficiency.

The dyeing tank 110, counterstain tank 120, dyeing-promoting tank 130, washing tank 140, and second washing tank 150 may be separate, i.e., all or part of them may be independently disposed and moved (if having a movement function). In addition, as shown in fig. 9 and 10, the grooves may be fixedly assembled into a whole, and integrally movable.

In this embodiment, as shown in fig. 10, the pickup assembly 210 is driven by a first driving assembly 220 (including an X-axis motor 221, a Y-axis motor 222, and a Z-axis motor 223) to move in XYZ three-axis directions, thereby moving, inserting, and picking and placing the slide 500. The width direction of the slide 500 inserted into the accommodating groove 100 (i.e., the width direction of the insertion region) is the X direction, the arrangement direction of the slide 500 inserted into the accommodating groove 100 (i.e., the thickness direction of the insertion region) is the Y direction, and the vertical direction, which is the insertion direction of the slide 100, is the Z direction.

Further, in one embodiment, the transport mechanism 200 includes a Y-axis track 2221 and Y-axis slide 2222 in the Y-direction, an X-axis track 2211 and X-axis slide 2212 in the X-direction, a Y-axis motor 222, and an X-axis motor 221. The X-axis motor 221 and the Y-axis motor 222 may be, for example, stepping motors, servo motors, or the like.

The Y-axis rail 2221 extends linearly in the Y-direction and is fixed to the lower surface of the support member 230. The support member 230 may be a top wall portion of a housing of the sample staining apparatus, a beam member for support, or the like. The Y-axis slider 2222 is mounted on the surface side of the lower side of the Y-axis rail 2221 and is movable along the Y-axis rail 2221. The Y-axis motor 222 moves the Y-axis slider 2222 in the Y-direction through the corresponding transmission mechanism. The transmission mechanism may be a belt-pulley mechanism, a rack-and-pinion mechanism, or the like.

The X-axis rail 2211 extends linearly in the X-direction and is fixed to the lower surface of the Y-axis slider 2222. The X-axis slider 2212 is mounted on the side of the face on the lower side of the X-axis track 2211 and is movable along the X-axis track 2211. The X-axis motor 221 moves the X-axis slider 2212 in the X-direction through a corresponding transmission mechanism.

The Y-axis slider 2222, the X-axis track 2211, the X-axis slider 2212, the X-axis motor 221, and the Y-axis motor 222 are each provided with a pair. A pair of X-axis slide blocks 2212 are mounted on one side of the underside thereof with pickup assembly 210. In this way, the pick-up assemblies 210 can be independently moved in the X direction along the respective X-axis tracks 2211. In addition, the pick-up assembly 210 can also independently move in the Y direction along the common Y-axis track 2221.

Further, in one embodiment, the first drive assembly 220 includes a Z-axis motor 223 for the pick-up assembly 210 (e.g., clamp 2232) and a transmission 2231. The Z-axis motor 223 lifts the grip 2232 via a transmission mechanism 2231. The transmission mechanism 2231 may be a belt pulley mechanism, a rack and pinion mechanism, or the like.

In addition, a drying tub 160 and an air supply unit 610 may be further included in some embodiments. The air supply unit 610 may generate an air flow for drying, and blow the slide 500 positioned in the drying bath 160, thereby achieving drying of the slide 500. The air supply unit 610 includes, for example, an electric fan, which can forcibly supply air into an air passage inside the drying tub 160. The slide 500 is placed on the second placing part 161 inside the drying groove 160, and the air supply unit 610 can dry the slide 500 by supplying air to the slide 500 for a certain time. By providing the drying bath 160 and the blower unit 610, the stained slide 500 can be quickly dried, and the time required for the staining process can be shortened. In addition, similarly, the slide 500 can be inserted into and extracted from the second mounting member 51 by the transfer mechanism 200, and therefore, the structure of the apparatus can be simplified.

In one embodiment, the air conditioner further includes a heater 620 for heating the air sent from the air supply unit 610. The heater 620 is located between the air supply unit 610 and the drying tub 160. The air supplied from the air supply unit 610 is supplied to the air passage in the drying tub 160 in a warm air state having a temperature rise after being heated by the heater 620. This can dry the stained slide 500 more quickly, and thus the time required for the staining process can be further shortened.

In the whole dyeing process, the first dye liquor and the second dye liquor can not lose efficacy in a short period, and the first dye liquor and the second dye liquor are repeatedly used. Considering the volatility of the first dye liquor, please refer to fig. 10, in one embodiment, the device further includes a first dye liquor path control system (not shown in detail in the drawing, but only indicated schematically by corresponding lines, and the related liquid path control systems described below are all indicated in this way) and a first dye liquor seal container 710, where the first dye liquor path control system is respectively in communication with the first dye liquor seal container 710 and the dyeing tank 110, and is configured to recycle the corresponding first dye liquor in the dyeing tank 110 to the first dye liquor seal container 710, and to re-drain the first dye liquor in the first dye liquor seal container 710 to the dyeing tank 110.

The first dye liquor path control system may include a first dye liquor recovery system and a first dye liquor inlet system, where the first dye liquor recovery system is respectively communicated with the first dye liquor sealing container 710 and the dyeing tank 110, and is used for recovering the corresponding first dye liquor in the dyeing tank 110 to the first dye liquor sealing container 710. The first dye liquor inlet system is respectively communicated with the first dye liquor sealing container 710 and the dyeing tank 110, and is used for discharging the first dye liquor in the first dye liquor sealing container 710 into the dyeing tank 110 again.

In the above embodiments, the first dye liquor recovery system and the first dye liquor feed system are used to implement recovery and feed liquor control, respectively. In other embodiments, the first dye liquor path control system may be an integral system having not only the recycling function but also the liquor feeding function.

In one embodiment, the first dye liquor recovery system includes a first dye liquor recovery line in communication with the dye vat 110 and the first dye liquor seal vessel 710, respectively, and a first dye liquor recovery pressure source in direct or indirect communication with the first dye liquor recovery line. The first dye liquor recovery pressure source pumps the first dye liquor in the dye vat 110 into the first dye liquor seal container 710.

In one embodiment, the first dye liquor feed system includes a first dye liquor feed line and a first dye liquor feed pressure source, the first dye liquor feed line being in communication with the dye vat 110 and the first dye liquor seal vessel 710, respectively, and the first dye liquor feed pressure source being in direct or indirect communication with the first dye liquor feed line. The first dye liquor inlet pressure source discharges the first dye liquor in the first dye liquor seal container 710 into the dyeing tank 110.

More commonly, the first dye liquor recovery pressure source and the first dye liquor inlet pressure source can both utilize air pressure to drive the flowing direction of the liquid, and other pressing modes are also feasible. In one embodiment, the first dye liquor recovery pipeline and the first dye liquor feed pipeline are the same pipeline, and/or the first dye liquor feed pressure source and the first dye liquor recovery pressure source are the same pressure source. For example, the first dye liquor recovery pipeline and the first dye liquor inlet pipeline are the same pipeline, and the recovery and liquid inlet functions are realized through connection with different pressure sources and cooperation with the opening and closing design of each pipeline.

In one embodiment, after the control unit determines that there is no sample to be dyed or receives the first dye liquor recycling instruction sent by the user, the first dye liquor path control system recycles the corresponding first dye liquor in the dyeing tank 110 to the first dye liquor sealing container 710.

When the control unit determines that the sample needs to be dyed or receives the first dye liquor inlet instruction sent by the user, the first dye liquor path control system re-discharges the first dye liquor in the first dye liquor sealing container 710 into the dyeing tank 110.

Since the first dye liquor can be reused, the first dye liquor sealing container 710 for storing the first dye liquor is not required to be oversized, and only the usage amount of each dyeing is required to be basically satisfied at least. At this time, the first dye liquor sealing container 710 may be a reagent bottle with a smaller volume, and a larger recycling bin or other container is not required. The reagent bottle can even be the original reagent bottle of the first dye liquor (if the original reagent bottle exists in the first dye liquor), so that when the first dye liquor needs to be replaced, the reagent bottle can be directly replaced without the need of discharging. When the reagent bottle is used as the first dye liquor sealing container 710, the volume is small, in one embodiment, the reagent bottle can be arranged in the smear preparation equipment, so that the reagent bottle not only can protect the first dye liquor sealing container 710, but also can ensure the compactness of the whole equipment due to the small volume of the reagent bottle. And adopt structures such as bigger recycling bin, be unfavorable for setting it in equipment, will increase the volume of equipment.

Since the first dye liquor usually uses methanol of high purity as solvent, it has very strong volatility, and it is volatilized quickly when exposed to air. The structure can recycle, seal and preserve the first dye liquor when not in use, so as to avoid volatilization and prolong the service life of the first dye liquor.

In one embodiment, the dyeing apparatus further comprises a first dye liquor cover, wherein the first dye liquor cover is used for shielding a corresponding opening on the dyeing tank 110 when the dyeing tank 110 is in a non-use state and re-opening the dyeing tank 110 when the dyeing tank is required to be used.

The first dye liquor cover can be controlled independently or together with the cover of other holding tanks (e.g. counterstain tank 120). The first bath cover may close the dyeing tank 110 when not in use, thereby avoiding volatilization of the first bath.

For the timing of recycling or capping, in one embodiment, the manner in which the control unit determines that no sample is required to perform the staining operation includes, but is not limited to: the staining tank 110 has no sample and no sample to be stained is fed in, and/or the staining tank 110 idle time exceeds a set value.

Further, for the first dye liquor, the first dye liquor can be reused, so that the replacement frequency is greatly reduced, frequent replacement is not needed, and the labor intensity of operators can be reduced. In the simplest form, the first dye liquor may be manually taken and placed by the user when it is required to be replaced.

Of course, in order to improve the working efficiency and further implement automation, in one embodiment, the dyeing machine further includes a first dye liquor discharging system, where the first dye liquor discharging system is communicated with the dyeing tank 110 or the first dye liquor sealing container 710, and when the control unit determines that the first dye liquor reaches the discharging requirement or after receiving the first dye liquor discharging instruction sent by the user, the first dye liquor discharging system discharges the first dye liquor in the dyeing tank 110 or the first dye liquor sealing container 710.

The discharge requirements of the first dye liquor may be set according to actual requirements, and in one embodiment, the discharge requirements include, but are not limited to: the ratio of the number of times of dyeing the sample to the volume of the first dye liquor is more than 2 times/ml and/or the service time of the first dye liquor is more than 2 days. For example, when 50ml of the first dye solution is stored in the dye tank 110, the first dye solution may be discharged after the sample is subjected to dyeing operation for 100 times or more in the first dye solution. The number of times of dyeing is not absolutely equal to the number of dyed samples, and when the samples complete a complete dyeing action in the first dye liquor to be dyed once, the same sample can be dyed for a plurality of times in the first dye liquor due to different dyeing requirements or dyeing effects. When the first dye liquor reaches the requirements, the first dye liquor can be manually or automatically discharged and replaced by a user. Because the dye liquor is generally high in cost, the cost can be greatly reduced by recycling the dye liquor.

Similar to the recycling of the first dye liquor, in one embodiment, the dye liquor recycling device further comprises a second dye liquor path control system and a second dye liquor recycling container 720, wherein the second dye liquor path control system is communicated with the second dye liquor sealing container and the counterstain 120, and is used for recycling the corresponding second dye liquor in the counterstain 120 to the second dye liquor recycling container 720 and re-discharging the second dye liquor in the second dye liquor recycling container 720 to the counterstain 120.

Likewise, the second dye liquor path control system may include a second dye liquor recovery system and a second dye liquor inlet system, where the second dye liquor recovery system is communicated with the second dye liquor recovery container 720 and the counterstain 120, and is used to recover the second dye liquor corresponding to the counterstain 120 to the second dye liquor recovery container 720; the second dye liquor inlet system is communicated with the second dye liquor recovery container 720 and the counterstain tank 120, and is used for discharging the second dye liquor in the second dye liquor recovery container 720 into the counterstain tank 120 again.

Of course, in some embodiments, the second dye liquor path control system may also be an integral system, which has not only the recycling function but also the liquor feeding function.

In one embodiment, the second dye liquor recovery system includes a second dye liquor recovery line and a second dye liquor recovery pressure source, the second dye liquor recovery line being in communication with the counterstain 120 and the second dye liquor recovery vessel 720, respectively, the second dye liquor recovery pressure source being in direct or indirect communication with the second dye liquor recovery line. The second liquor recovery pressure source pumps the second liquor in the counterstain 120 into the second liquor recovery vessel 720.

The second dye liquor inlet system comprises a second dye liquor inlet pipeline and a second dye liquor inlet pressure source, the second dye liquor inlet pipeline is respectively communicated with the counterstain 120 and the second dye liquor recovery container 720, and the second dye liquor inlet pressure source discharges the second dye liquor in the second dye liquor recovery container 720 into the counterstain 120.

The second dye liquor recovery pressure source and the second dye liquor feed liquid pressure source can be set by referring to the first dye liquor recovery pressure source and the first dye liquor feed liquid pressure source. In one embodiment, the second dye liquor recovery pipeline and the second dye liquor feed liquor pipeline are the same pipeline, and/or the second dye liquor feed liquor pressure source and the second dye liquor recovery pressure source are the same pressure source.

The timing of the second dye liquor recovery and liquor feed may include, but is not limited to, the following:

When the control unit determines that no sample needs to perform the counterstaining operation or receives a second dye liquor recovery instruction sent by the user, the second dye liquor path control system recovers the second dye liquor corresponding to the counterstaining tank 120 to the second dye liquor recovery container 720.

When the control unit judges that the sample needs to be subjected to the counterstaining operation or receives a second dye liquor inlet instruction sent by a user, the second dye liquor path control system discharges the second dye liquor in the second dye liquor recovery container 720 into the counterstaining groove 120 again.

Likewise, in addition to recycling, sealing and preserving, in an embodiment, a second dye liquor cover may be further included, and when the control unit determines that no sample needs to perform the counterstaining operation or receives the instruction of sealing the second dye liquor from the user, the second dye liquor cover is controlled to cover the counterstaining tank 120; when the control unit judges that the sample needs to be subjected to the counterstaining work or receives a second dye liquor opening instruction sent by a user, the control unit controls the second dye liquor cover to open the counterstaining groove 120.

Likewise, for the dyeing promotion tank 130, the cleaning tank, and the second cleaning tank 150, the recovery and feeding of the liquid therein can also be achieved with reference to the recovery and feeding system described above.

The recovery and feed systems corresponding to the dyeing promoting tank 130, the cleaning tank 140 and the second cleaning tank 150 can be implemented by adopting the same structure as the first dyeing liquor path control system and the first dyeing liquor sealing container 710. For example, the dyepromoting tank 130 communicates with the liquor recovery vessel 730 via a liquor circuit control system to effect recovery and liquor feed. The cleaning tank 140 is communicated with the cleaning solution recovery container 740 through a cleaning solution path control system, so that recovery and liquid feeding are realized. The second cleaning tank 150 is communicated with the second cleaning liquid recovery container 750 through a second cleaning liquid path control system, so that recovery and liquid feeding are realized.

Of course, the dyeing acceleration tank 130, the cleaning tank 140 and the second cleaning tank 150 may be sealed with corresponding covers.

In one embodiment, the device further comprises an dyeing promoting liquid cover, and when the control unit judges that no sample needs to be subjected to the dyeing promoting work or receives a cover dyeing promoting liquid instruction sent by a user, the control unit controls the dyeing promoting liquid cover to cover a corresponding opening on the dyeing promoting tank 130 and reopens the dyeing promoting tank 130 when the sample needs to be used.

In one embodiment, the cleaning device further comprises a cleaning liquid cover, and when the control unit determines that no sample needs to be subjected to the cleaning operation before counterstaining or receives a cleaning liquid sealing instruction sent by a user, the control unit controls the cleaning liquid cover to cover a corresponding opening on the cleaning tank 140 and reopens the cleaning tank 140 when the sample needs to be used.

In one embodiment, the second cleaning tank 150 may also be provided with a corresponding second cleaning tank cover, and the second cleaning tank 150 is sealed under the control of the control unit.

The recovery and liquid inlet system and the corresponding cover body corresponding to each accommodating groove can select one or more of the accommodating grooves for use according to actual requirements, and generally, the first dye liquid and the second dye liquid can be recycled preferentially so as to reduce dyeing cost.

The liquid inlet system and the recovery system corresponding to the dyeing tank 110, the counterstain tank 120, the dyeing promoting tank 130, the cleaning tank 140 and the second cleaning tank 150 can be used alternatively or simultaneously. The recovery pressure source and the feed liquid pressure source can be respectively arranged, and the same pressure source can also be adopted as the pressure sources of suction and feed liquid.

In the case where the above-mentioned recovery system is provided, each liquid may be recovered and stored after the completion of the dyeing work every day (or each batch of samples), and then released into the corresponding accommodation groove 100 when it is used next time. Of course, after the liquid is continuously used for a period of time, the liquid can be recycled and stored, so that the volatilization amount of the reagent is reduced, and the pollution of the reagent is avoided. The reagent in each of the containers 100 may be replaced periodically or after staining a quantitative sample.

Further, referring to fig. 9, in addition to providing a separate cover, in one embodiment, the sample staining apparatus further includes an integral cover 300, and the integral cover 300 is used to cover the corresponding opening on each of the accommodating grooves 100 when the accommodating grooves 100 are in the non-use state. The non-use state may refer to a non-operation state, that is, when dyeing is not performed, or when a slide is not placed in one of the accommodating grooves 100. The integral cover 300 can cover all the receiving grooves 100 in a unified manner, for example, when a slide is not placed in a certain receiving groove 100, or when the corresponding receiving groove is covered, as indicated above, the integral cover 300 can cover the receiving groove 100.

Of course, the whole outer cover 300 can be manually covered on the accommodating groove 100 by a worker, or a transmission structure can be designed to automatically cover the accommodating groove 100.

In one embodiment, referring to fig. 9, the sample dyeing apparatus further includes a mounting base 400 and a second driving assembly (not shown), the accommodating groove 100 is mounted on the mounting base 400, and either one of the integral outer cover 300 and the mounting base 400 is in transmission connection with the second driving assembly, so as to enable the integral outer cover 300 to relatively move with the accommodating groove 100 on the mounting base 400 under the driving of the second driving assembly, and cover the corresponding opening on each accommodating groove 100.

In one embodiment, the receiving slot 100 is moved so that the integral cover 300 can cover the receiving slot 100. Wherein, each tank body in the accommodating tank 100 can move independently or integrally.

In fig. 9, the second driving assembly is used to drive the mounting base 400, and the integral cover 300 can be kept different, which is located above the corresponding opening of each receiving groove 100. The second drive assembly may drive the mount 400 to move along the x-axis. In operation, the integral cover 300 opens the corresponding opening of the receiving groove 100. When the accommodating groove 100 needs to be shielded, the second driving assembly drives the mounting seat 400 to communicate with the accommodating groove 100 to move towards the integral outer cover 300 until the integral outer cover 300 shields the corresponding opening of the accommodating groove 100.

The sample dyeing device can be used to implement the steps of the dyeing method, and the dyeing method is not limited to the device in this embodiment.

On the other hand, the application also provides a dyeing liquid combination for dyeing a sample, which comprises a first dyeing liquid and a second dyeing liquid, wherein the first dyeing liquid is a mixture of a first biological dye and a second biological dye, the first biological dye can dye acidophilic substances in the sample, and the second biological dye can dye alkalophilic substances in the sample. The second dye solution is a mixture of the third biological dye and the buffer solution, that is, most of the effective components in the second dye solution are the buffer solution and the third biological dye. The third biological dye is capable of staining the sample for an alkalophilic or acidophilic substance, which may be the same as or different from the first biological dye or the second biological dye. The first, second and third biological dyes may be selected from the dyes described above.

In some embodiments, the second dye solution may be a mixture of the first biological dye, the third biological dye, and the buffer. For example, jimsa dye liquor consists of azure (formed by oxidation of methylene blue) as the third biological dye and eosin as the first biological dye, but the amount of azure is significantly greater than the amount of eosin. The volume ratio of the first bio-dye is much smaller than the volume ratio of the third bio-dye, e.g. the ratio of the first bio-dye to the third bio-dye may be less than 1/3.

The staining solution combination can be used for, but is not limited to, the staining method and the sample staining device, and can also be used for other staining methods and sample staining devices.

As a more specific example, the first biological dye is eosin dye, the second biological dye is methylene blue dye, and the third biological dye is azure dye. Of course, the first, second and third biological dyes may be replaced with other dyes that are capable of dyeing acidophilic and alkalophilic substances, respectively.

Further, to meet the usage habits of most clinical departments, in one embodiment, the first dye liquor is a Rui-Jiemsa dye liquor, which includes eosin dye and methylene blue dye. The second dye solution is a mixture of Giemsa dye solution and buffer solution, and the Giemsa dye solution contains methylene blue dye as a third biological dye.

The foregoing description of the application has been presented for purposes of illustration and description, and is not intended to be limiting. Variations of the above embodiments may be made by those of ordinary skill in the art in light of the present teachings.

Claims

1. A sample dyeing method, characterized in that the sample dyeing method is applied to a smear preparation device, the smear preparation device comprises a containing groove, a conveying mechanism and a control unit, wherein the containing groove comprises a dyeing groove for storing a first dye liquor and a counterstain groove for storing a second dye liquor different from the first dye liquor; the sample staining method comprises the following steps:

Dyeing: the control unit controls the conveying mechanism to soak a sample in a dyeing tank and then take out the sample, wherein the dyeing tank stores a first dye liquor, the first dye liquor is Rui-Jiemsa dye liquor, the Rui-Jiemsa dye liquor is a mixture of a first biological dye and a second biological dye, the first biological dye is eosin dye, and the second biological dye is methylene blue dye;

And (3) counterstaining: the control unit controls the conveying mechanism to soak a sample in a counterstain tank and then take out the sample, the counterstain tank stores a second dye liquor, and the second dye liquor is different from the first dye liquor; wherein the second dye solution is a mixture of a third biological dye which is the only biological dye and a buffer solution, and the third biological dye can dye an alkalophilic substance or an acidophilic substance in a sample; or the second dye liquor also comprises a first biological dye, a third biological dye and a buffer solution, wherein the third biological dye is an alkaline biological dye, and the volume ratio of the first biological dye is far smaller than that of the third biological dye;

The method further comprises the steps of: the control unit controls the conveying mechanism to select the next slide smeared with the sample, and the dyeing step is repeated until the counterstaining step.

2. The method of staining a sample of claim 1, further comprising the step of dyeing: immersing the sample treated by the dyeing step in a dyeing promoting tank, and taking out the sample, wherein a buffer solution is stored in the dyeing promoting tank;

3. The method of staining a sample according to claim 2 wherein in the step of staining promotion, the sample is immersed in the buffer for a period of time ranging from 1 to 5 minutes.

4. The method of staining a sample of claim 2, further comprising the step of pre-counterstaining washing: and immersing the sample treated by the dyeing promotion step in a cleaning tank, and taking out the sample, wherein cleaning liquid is stored in the cleaning tank.

5. The method of staining a sample according to claim 1, further comprising the step of washing prior to counterstaining: washing the sample with a cleaning liquid, the cleaning liquid being stored in a washing tank;

6. The method of staining a sample according to claim 4 wherein the sample is washed for a period of time ranging from 1 to 5 minutes in the pre-counterstain washing step.

7. The sample staining method of claim 1, wherein the third biological dye is azure dye.

8. The method of claim 1, wherein the second dye solution is a mixture of giemsa dye solution and buffer solution, or the second dye solution is a mixture of jew B dye solution and buffer solution.

9. The method of staining a sample according to claim 8 wherein the volume ratio a of the giemsa or the liu B dye to the buffer in the second dye solution is 1/30.ltoreq.a.ltoreq.1/10.

10. The method of staining a sample according to claim 1 wherein the sample soaking time in the counterstaining step is from 5 to 15 minutes.

11. The method of staining a sample according to claim 1 wherein the sample soaking time is 1 to 5 minutes in the staining step.

12. The method of staining a sample of claim 1, further comprising a second washing step: and cleaning the sample treated by the counterstaining step.

13. The method of staining a sample of claim 1, further comprising:

14. The method of staining a sample of claim 1, further comprising:

15. The method of staining a sample according to claim 13 wherein the control unit determining that no sample is required for the staining step comprises: no sample is in the dyeing tank and no sample is waiting to be sent in, and/or the idle time of the dyeing tank exceeds a set value.

16. The sample dyeing method according to claim 1, wherein the first dye liquor is discharged after the control unit judges that the first dye liquor reaches a set discharge requirement or receives a first dye liquor discharge instruction issued by a user.

17. The method of dyeing a sample according to claim 16, characterized in that the discharge requirements include a number of dyeings per milliliter of the first dye solution greater than or equal to 2 and/or a use time of the first dye solution of up to more than 2 days.

18. The method of staining a sample of claim 1, further comprising:

19. The method of staining a sample of claim 1, further comprising:

20. The method of staining a sample according to claim 18 wherein the means for determining that no sample is required for the counterstaining step comprises: and the counterstain tank and the dyeing tank are both free of samples and no samples are to be fed, and/or the idle time of the counterstain tank exceeds a set value.

21. The sample dyeing method according to claim 1, wherein the second dye liquor is discharged after the control unit judges that the second dye liquor reaches a set discharge requirement or receives a second dye liquor discharge instruction issued by a user.

22. The method of staining a sample of claim 4, further comprising:

23. The method of staining a sample of claim 2, further comprising:

24. A combination of dye liquors for dyeing a sample, comprising a first dye liquor and a second dye liquor, wherein the first dye liquor is a rui-giemsa dye liquor, the rui-giemsa dye liquor is a mixture of a first biological dye and a second biological dye, the first biological dye is eosin dye, the second biological dye is methylene blue dye, the second dye liquor is a mixture of a third biological dye which is the sole biological dye and a buffer solution, and the third biological dye is capable of dyeing an alkalophilic substance or an acidophilic substance in the sample; or the second dye liquor also comprises a first biological dye, a third biological dye and a buffer solution, wherein the third biological dye is an alkaline biological dye, and the volume ratio of the first biological dye is far smaller than that of the third biological dye.

25. The combination of claim 24, wherein the first biological dye is eosin dye, the second biological dye is methylene blue dye, and the third biological dye is azure dye.

26. The combination of claim 25, wherein the first dye liquor is a rui-giemsa dye liquor, the second dye liquor is a mixture of giemsa dye liquor and buffer liquor, or the second dye liquor is a mixture of a liu B dye liquor and buffer liquor.

27. The combination of staining fluids according to any one of claims 24 to 26, wherein in the second staining fluid the volume ratio a of giemsa staining fluid or lium B staining fluid to buffer fluid has a value of 1/30.ltoreq.a.ltoreq.1/10.

28. The combination of claim 24-26, wherein the second and third biological dyes are the same biological dye.