JP3620013B2 - Method and apparatus for staining blood smear - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a blood smear staining method and staining apparatus.
[0002]
[Prior art]
In hospitals and laboratories, in order to observe blood under a microscope, blood is smeared on a slide glass and a blood smear that is stained is used. In order to make a better observation, the blood must be stained appropriately.
[0003]
However, the conventional dyeing process is not managed, and the dyeing conditions such as the temperature and the amount of liquid differ from time to time, and stable dyeing results cannot be obtained. For this reason, the size of the cells in the sample blood, the size of the nuclei, and the hue differed greatly depending on the staining.
[0004]
[Problems to be solved by the invention]
As described above, since the process of conventional blood smear staining is not managed, there is a drawback in that the observation result varies greatly depending on the staining.
[0005]
The present invention has been made to solve such conventional drawbacks. An object of the present invention is to enable stable staining of a blood smear.
[0006]
[Means for Solving the Problems]
In the present invention, blood to be smeared on a slide glass, dried, immersed in a fixing solution in a fixing solution tank, immersed in a staining solution in a staining solution tank, immersed in a cleaning solution in a cleaning solution tank, and dried In the staining process of smears, the drying time after smearing, the drying temperature, the fixing solution soaking time , the fixing solution soiling degree, the staining solution soaking time , the staining solution concentration in the staining solution tank, and the stain of the cleaning solution Of these, at least the drying time and the drying temperature in the drying after smearing, and the time for dipping in the fixing solution are controlled among the drying time and the drying temperature in the drying after washing . When the dyeing process is managed in this way, a stable dyeing process is performed.
[0007]
If the drying setting time in the first drying is 1 minute or more and less than 15 minutes and the drying setting temperature is 35 ° C. or more and 42 ° C., a suitable dyeing result can be obtained.
[0008]
If the fixing solution is methanol and the immersion setting time of the fixing solution is between 10 and 30 seconds, a suitable staining result can be obtained.
[0009]
In the management of the degree of contamination of the fixing solution, the concentration of the staining solution, or the degree of contamination of the cleaning solution, if the absorbance of each solution is detected, the contamination and concentration can be accurately detected.
[0010]
The dyeing is light dyeing, and the dyeing liquid tank is composed of a first tank containing a stock solution of the light liquid and a second tank containing a dilute solution of the light liquid, and manages the immersion time in each tank. In this case, the set time is 2 minutes or more for the first tank and about 5 minutes for the second tank. Thereby, suitable light dyeing is performed.
[0011]
The staining may be Meigrunwald Giemsa staining.
[0012]
When the drying time and the drying time in the subsequent drying are managed, a preferable dyeing result can be obtained if the set time is about 5 minutes and the set temperature is about 37 degrees.
[0013]
DETAILED DESCRIPTION OF THE INVENTION
A staining apparatus according to an embodiment of the present invention will be described. FIG. 1 is a diagram showing the overall configuration. Each part will be described below.
[0014]
The conveyance part 10 is equipped with a conveyance arm in each place, and conveys a slide glass between each unit and in each unit.
[0015]
The smearing unit 11 is a unit that smears sample blood on a slide glass. The first drying unit 12 is a unit that dries the blood smeared on the slide glass. In FIG. 2, the structure of the smearing unit 11 and the 1st drying unit 12 is shown concretely. The smear unit 11 includes a dropping device 41 that sucks the blood 2 stored in the container 3 and drops the blood 2 on the slide glass 1, a drawing glass 43, and a drawing glass moving device 44 that moves the drawing glass 43. Yes. The first drying unit 12 includes a 37 ° constant temperature plate 45, a fan 46, and a temperature sensor 21 that detects the temperature in the first drying unit 12. Here, the transport unit 10 includes a transport arm 42 that transports the slide glass 1.
[0016]
The fixing unit 13 shown in FIG. 1 is a unit that fixes the blood 2 to the slide glass 1, the first staining unit 14 is a unit that performs a staining process with the first staining solution, and the second staining unit 15 is the first staining unit 15. The washing unit 16 is a unit that performs washing with a washing solution, and the second drying unit 17 is a unit that performs drying after washing by the washing unit 16. FIG. 3 shows a specific example of an apparatus composed of these units. Each unit will be described with reference to FIG.
[0017]
The fixing unit 13 includes a fixing liquid tank 51 that stores the fixing liquid, and includes a liquid level sensor 22 that detects the amount of the fixing liquid, and an absorbance sensor 23 that detects contamination of the fixing liquid. Methanol is used as the fixing solution. The absorbance sensor 23 includes, for example, a light emitting element and a light receiving element. The light receiving element receives light from the light emitting element that has passed through the liquid to be measured, and detects the absorbance of the liquid based on the strength of the output signal.
[0018]
The first staining unit 14 includes a first staining liquid tank 52. The first dyeing tank 52 contains a stock solution of light liquid and is provided with a liquid level sensor 24.
[0019]
The second staining unit 15 includes a second staining liquid tank 53. The second staining tank 53 contains a diluted solution of light liquid, and includes an absorbance sensor 26 for detecting the concentration of the liquid and a liquid level sensor 25 for detecting the amount of the liquid.
[0020]
The cleaning unit 16 includes a cleaning liquid tank 54. Here, the cleaning liquid tank 54 a mixture of distilled water and phosphate buffer is contained as a cleaning liquid, and a pH sensor 27 for detecting the pH of the washing solution, the absorbance sensor 28 for detecting the contamination of the cleaning liquid .
[0021]
The second drying unit 17 includes a 37 ° hot air fan 55 and a temperature sensor 29 that detects the temperature in the unit.
[0022]
The control unit 31 illustrated in FIG. 1 controls the transport unit 10, the smear unit 11, the first drying unit 12, and the second drying unit 17. Each of the sensors 21 to 29 is connected to the control unit 31. If any one of these detected values becomes an abnormal value, the alarm generation unit 32 issues an alarm indicating which one has become abnormal. It is supposed to be generated. The control part 31, the conveyance part 10, the alarm generation part 32, and the sensors 21-29 comprise a management means.
[0023]
Next, the operation of the apparatus configured as described above will be described with reference to the flowchart of FIG.
[0024]
First, in step S1, blood 2 is smeared on the slide glass 1. Specifically: In the smear unit 11, when the operator sets the sample blood 2 accommodated in the container 3 and the ride glass 1 at predetermined positions, the dropping device 41 drops the sample blood 2 onto the slide glass 1. Next, the transfer arm 42 places the slide glass 1 on which the blood 2 is dropped at a predetermined position. Therefore, the drawing glass 43 extends the blood 2 (see the parentheses in FIG. 2).
[0025]
In the next step S2, blood 2 smeared on the slide glass 1 is dried. Specifically: The transport arm 42 transports the slide glass 1 smeared with blood 2 (hereinafter referred to as a specimen A for the sake of simplicity) to the first drying unit 12 and places it on a 37-degree thermostatic plate 45. After the minute has elapsed, the temperature is lowered from the thermostat 45. Next, the fan 46 is started to operate, and the specimen A is dried by blowing air.
[0026]
Here, the time for placing on the thermostat 45 and the temperature of the thermostat 45 will be described. It may take some time to dry when the humidity is high in summer or cold in winter. However, such a situation can be accommodated by drying for 1 minute at a temperature of 37 degrees. There is no problem if it is in the range from 35 degrees to 42 degrees within 1 minute to less than 15 minutes. If it is 35 degrees or less, drying in 1 minute may be insufficient, and if it is 42 degrees or more, the blood cells are broken or bluish. In addition, if the drying time is set to 15 minutes or more, it is not good because it causes the dyed color to vary.
[0027]
In the next step S3, the blood 2 smeared on the slide glass 1 is fixed with methanol. That is, in the fixing unit 13, the specimen A is immersed in the fixing liquid in the fixing liquid tank 51. The immersion time is optimally 20 seconds, but there is no particular problem if it is between 10 seconds and 30 minutes. In addition, after this step, a plurality of specimens A are collectively transported by a transport arm 58 as shown in FIG.
[0028]
In the next step S4, a first staining process is performed. That is, in the first staining unit 14, the specimen A is immersed in the staining liquid in the first staining liquid tank 52 for 2 minutes.
[0029]
In the next step S5, a second staining process is performed. That is, in the second staining unit 15, the specimen A is immersed in the staining liquid in the second staining liquid tank 53 for 5 minutes.
[0030]
As described above, in the first dyeing unit 14 and the second dyeing unit 15, the immersion time was set to 2 minutes and 5 minutes, respectively, but these are the concentrations of the respective solutions in the first dye solution tank 52 and the second dye solution tank 53. It is determined corresponding to In this example, the first staining solution is a light stock solution containing light dye in 100% methanol, and the second staining solution is a phosphate buffer solution having a pH of 7.0 and a light stock solution added at a rate of 10%. is there. In this case, by setting the immersion time of the first staining liquid tank 52 to 2 minutes or longer, the influence of unevenness due to individual differences or the like on the dyed shade in the staining in the second staining liquid tank 53 is eliminated. The longer the immersion time of the second dye bath 53 is, the deeper the dyeing is, and the immersion time is determined by the practitioner so as to be dyed to a desired shade. A dyeing of about 5 minutes is the most beautiful.
[0031]
In the next step S6, the specimen A is washed. That is, the specimen A is immersed in the cleaning liquid in the cleaning liquid tank 54 in the cleaning unit 16. As this cleaning solution, a solution obtained by adding a phosphate buffer to distilled water at a rate of 10% is used. Here, the phosphate buffer is an aqueous solution of hydrogen phosphate 2Na and dihydrogen phosphate K. The same applies to the phosphate buffer used in the second staining solution.
[0032]
In the next step S7, the specimen A is dried. That is, the specimen A is transported to the second drying unit 17 and dried by the hot air fan 55. Here, the optimum temperature is about 37 degrees and the drying time is about 5 minutes.
[0033]
In the course of the above processing, the control unit 31 monitors the signals sent from the sensors 21 to 28, and when those signal values deviate from the set values, signals indicating the respective abnormalities are sent to the alarm generation unit. 32. That is,
When the drying temperature of the first drying unit 12 deviates from the set temperature,
If the amount of fixative in the fixed unit 13 deviates from the set amount,
When the absorbance of the fixing solution of the fixing unit 13 deviates from the set absorbance,
When the amount of the first staining liquid in the first staining unit 14 deviates from the set amount,
When the absorbance (concentration) of the second staining solution of the second staining unit 15 deviates from the set absorbance,
When the amount of the second staining liquid in the second staining unit 15 deviates from the set amount,
When the pH of the cleaning liquid of the cleaning unit 16 deviates from the set pH value,
If the absorbance of the cleaning liquid in the cleaning unit 16 deviates from the set absorbance,
When the drying temperature of the second drying unit 17 deviates from the set drying temperature,
In each of the cases, an alarm indicating that the alarm is generated from the alarm generation unit 32.
[0034]
When an alarm occurs, the operator performs processing corresponding to each. If any of the liquid volume, concentration, pH , and absorbance detected by the sensors 22 to 28 is abnormal, the entire liquid in which the abnormality is detected is replaced. The management of each solution is described below.
[0035]
For the methanol fixative, manage the contamination of the solution. Here, the amount of liquid decreases because the fixing liquid is brought into the next tank. When the amount of the liquid becomes equal to or less than the set value, the liquid level sensor 22 detects this, and an alarm is generated assuming that the contamination has occurred, and the operator is notified. Further, when the fixing solution becomes dirty by processing a number of specimens A, it is monitored by the absorbance sensor 23. When the transmittance decreases to a set value, an alarm is generated as the fixing solution becomes dirty, Notify the operator.
[0036]
For the first staining liquid, the liquid volume is managed. Since this liquid is brought into the next tank, and since the solvent of this liquid is 100% methanol and has high volatility, it is greatly reduced when used for a long time. This is detected by the liquid level sensor 24, and when it becomes less than the set value, an alarm is generated to notify the operator.
[0037]
About the 2nd dyeing | staining liquid, a density | concentration and a liquid quantity are managed. Since the concentration of this liquid changes from the previous tank, it is detected by the absorbance sensor 26. When the concentration exceeds the set value, an alarm is generated to notify the operator. Further, when the amount of liquid is reduced from the set amount by bringing it into the next tank, it is detected by the liquid level sensor 26 and an alarm is generated to notify the operator.
[0038]
For cleaning fluids, control pH and soiling. The pH is controlled for the following reason. The pH greatly affects dyeing ( pH 6.4 to 7.0 is optimal), but the hue changes depending on the pH of the washing solution. The pH of distilled water is not very stable. Therefore, a buffer solution was added to the distilled water to stabilize the pH of the distilled water so that the dyeing was constant. The pH is detected by a pH sensor 27. In addition, since it is a solution for washing out the staining solution, it will become dirty and colored when used for a long time. This is detected by the absorbance sensor 28. When the absorbance exceeds a set value, an alarm is generated to notify the operator.
[0039]
When the drying temperature becomes abnormal in each of the first drying unit 12 and the second drying unit 17, the operator performs an operation for removing the cause of the abnormality.
[0040]
In the above embodiment, dyeing with a light solution is performed. This dyeing process may be performed by other dyeing methods. For example, in the Maygrun Wald Giemsa staining method, the staining unit is as shown in FIG. That is, there are three types of staining liquids, and the staining liquid tanks are a first staining liquid tank 61 in which a Meygurnwald stock solution is stored, a second staining liquid tank 62 in which a 10% Meygurnwald liquid is stored, and 3 And a third staining solution tank 63 containing% Giemsa solution. In this example, absorbance sensors 71 and 72 for detecting the absorbance of the liquid are provided in the second staining liquid tank 62 and the third staining liquid tank 63, respectively, thereby detecting the concentration of each liquid. In addition, liquid level sensors 73, 74, and 75 for detecting the liquid level are provided in each of the first staining liquid tank 61, the second staining liquid tank 62, and the third staining liquid tank 63, thereby detecting the amount of each liquid. Like to do. Other configurations are the same as those of the above-described embodiment.
[0041]
In the above embodiment, the smear unit 11 may be smeared using a centrifugal rotating device 81 as shown in FIG. Other configurations are the same as those of the above-described embodiment.
[0042]
【The invention's effect】
According to the present invention, since the processing conditions are managed in the staining process, a blood smear subjected to a stable staining process can be obtained. For this reason, the size of cells in the blood, the size of nuclei, and the hue do not vary greatly by staining, and accurate diagnosis can be performed.
[Brief description of the drawings]
FIG. 1 is a diagram showing an overall configuration of a specimen staining apparatus according to the present invention.
FIG. 2 is a diagram showing a configuration of a smearing unit and a first drying unit shown in FIG.
FIG. 3 is a view showing an apparatus including the fixing unit, the first and second staining units, the cleaning unit, and the second drying unit shown in FIG. 1;
4 is a diagram showing a flow of a staining process by the specimen staining apparatus shown in FIG. 1. FIG.
FIG. 5 is a view for explaining the device of the present invention when the Meigrun Wald Giemsa staining method is used.
FIG. 6 is a view showing another example of a smearing unit.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 10 Conveyance part 11 Smearing unit 12 1st drying unit 13 Fixed unit 14 1st dyeing unit 15 2nd dyeing unit 16 Cleaning unit 17 2nd drying units 21-29 Sensor 31 Control part 32 Alarm generating part

Claims (22)

A first step of smearing sample blood on a slide glass;
A second step of drying the sample blood smeared on the slide glass;
A third step of immersing the sample blood smeared on the slide glass in a fixative in a fixative bath;
A fourth step of immersing the sample blood smeared on the slide glass in a staining solution in a staining solution tank;
A fifth step of immersing the sample blood smeared on the slide glass in a cleaning solution in a cleaning solution tank;
In the method for staining a blood smear, which sequentially performs each of the steps of drying the sample blood smeared on the slide glass,
The drying time in the second step is set to a time of 1 minute or more and less than 15 minutes, the drying temperature is set to a temperature of 35 degrees or more and 42 degrees, and each is managed.
A method for staining a blood smear, wherein the immersion time in the third step is set to a time of 10 to 30 seconds and managed . The method of staining a blood smear according to claim 1 , wherein the concentration of the staining solution in the fourth step is managed based on an absorbance detected by an absorbance sensor provided in the staining solution tank. . The method for staining a blood smear according to claim 1 or 2, wherein the fixing solution in the third step is methanol. The blood smear according to any one of claims 1 to 3 , wherein the stain level of the fixative is managed based on absorbance detected by an absorbance sensor provided in the fixative tank. Dyeing method. Dyeing method of a blood smear according to any one of claims 1 to 4, characterized in that dyeing in the fourth step is Wright's stain. 6. The method for staining a blood smear according to claim 5, wherein the staining liquid tank comprises a first tank in which a stock solution of light liquid is accommodated and a second tank in which a dilution liquid of light liquid is accommodated. The method for staining a blood smear according to claim 6 , wherein the immersion time in the fourth step is set to 2 minutes or more in the first tank for management . The method of staining a blood smear according to claim 7 , wherein the immersion time in the fourth step is set to about 5 minutes in the second tank and managed . The method for staining a blood smear according to any one of claims 1 to 4 , wherein the staining in the fourth step is a Meigrunwald Giemsa staining. The stain of the blood smear according to any one of claims 1 to 9 , wherein the degree of contamination of the cleaning liquid is managed based on an absorbance detected by an absorbance sensor provided in the cleaning liquid tank. Method. The blood smear according to any one of claims 1 to 10 , wherein the drying time in the sixth step is set to about 5 minutes and the drying temperature is set to about 37 degrees to manage each. Dyeing method. Smearing means for smearing the sample blood on the slide glass;
First drying means for drying the sample blood smeared on the slide glass by the smearing means;
Fixing means for immersing the sample blood smeared on the slide glass in a fixing solution in a fixing solution tank;
Staining means for immersing the sample blood smeared on the slide glass in a staining solution in a staining solution tank;
A cleaning means for immersing the sample blood smeared on the slide glass in a cleaning liquid in a cleaning liquid tank;
A second smearing device for drying the sample blood smeared on the slide glass;
A management means for managing a drying time and a drying temperature in the first drying means and an immersion time in the fixing means;
The management means manages the set drying time in the first drying means as a time of 1 minute or more and less than 15 minutes, manages the set drying temperature as a temperature of 35 degrees or more and 42 degrees, and sets the set immersion time in the fixing means. Is managed as a time between 10 and 30 seconds . The blood smear staining apparatus according to claim 12 , wherein the management means further manages the concentration of the staining liquid based on an absorbance detected by an absorbance sensor provided in the staining liquid tank. . Dyeing apparatus of the blood smear according to any crab of claim 12 or claim 13, wherein the fixative in the fixing means is methanol. Said management means further contamination degree of the fixative, in any one of claims 12 to claim 14, characterized in that managed on the basis of the absorbance is detected by absorbance sensor provided in said stationary vessel The blood smear staining apparatus as described. The blood smear staining apparatus according to any one of claims 12 to 15 , wherein the staining liquid is a light staining liquid. The staining apparatus for blood smears according to claim 16 , wherein the staining liquid tank comprises a first tank for storing a stock solution of light liquid and two tanks for storing a diluted liquid of light liquid. 18. The blood smear staining apparatus according to claim 17 , wherein the management means further manages the immersion time of the first tank by setting it to 2 minutes or more . The blood smear staining apparatus according to claim 18 , wherein the management means further manages the immersion time of the second tank by setting it to about 5 minutes . The blood smear staining apparatus according to any one of claims 12 to 15 , wherein the staining liquid is a staining liquid for performing Meigrunwald Giemsa staining. It said management means further describes a dust concentration of the cleaning liquid, in any one of claims 12 to claim 20, characterized in that managed on the basis of the absorbance is detected by absorbance sensor provided in the washing liquid tank Blood smear staining device. The management unit is further configured to set the drying time in the second drying means in about 5 minutes, more of claims 12 to claim 21, characterized in that respectively manage by setting the drying temperature to about 37 degrees A blood smear staining apparatus according to claim 1.

Images