JP4068792B2 - Observation specimen preparation method and apparatus - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a method for producing an observation specimen (preparation) used for observing a thin slice sample cut from a biological sample or the like for the purpose of observing, examining, and analyzing a tissue such as a biological sample, animals and plants, and food. In particular, the present invention relates to a method suitable for automating the work of attaching and fixing a thin section sample on a slide glass.
[0002]
[Prior art]
7A to 7C show an outline of a conventional method for producing a specimen for observation (preparation).
[0003]
First, a sample block in which a specimen (for example, a biological sample) is embedded in an embedding material (for example, paraffin) is prepared. The surface layer portion of the sample block is sliced with a cutter (not shown), and the sliced slice sample 1 is adsorbed to the lower surface of the carrier tape 2 by the electrostatic force and taken out.
[0004]
Next, as shown in FIG. 7 a, the carrier tape 2 is run to send the thin slice sample 1 to the upper side of the slide glass 3 and stop there. Note that water (or water containing about 10 to 30 wt% of ethyl alcohol) is applied as an adhesive liquid 4 in advance on the slide glass 3.
[0005]
Next, as shown in FIG. 7 b, the carrier tape 2 is pushed downward to bring the thin slice sample 1 into close contact with the upper surface of the slide glass 3.
[0006]
Next, as shown in FIG. 7c, the carrier tape 2 is pulled upward. The thin slice sample 1 adheres to the upper surface of the slide glass 3 by the adhesive liquid 4 and is transferred onto the slide glass 3.
[0007]
Next, the slide glass 3 on which the thin section sample 1 is placed is placed on a hot plate (not shown) and held at a temperature of 40 to 50 ° C. for about 30 minutes, for example. As a result, the wrinkles or curls (rounds) generated in the thin slice sample 1 during thin cutting are stretched and the adhesive solution is evaporated to fix the thin slice sample 1 on the slide glass 3.
[0008]
(Problems of conventional methods)
Usually, wrinkles or curls are generated in the thin slice sample taken out by being adsorbed on the lower surface of the carrier tape. Such wrinkles or curls are stretched and removed by heating the slide glass and evaporating the adhesive liquid as described above. However, when wrinkles or curls are relatively large, this method cannot be completely removed and may remain in the prepared observation specimen.
[0009]
[Problems to be solved by the invention]
The present invention has been made in view of the problems of the conventional method for preparing an observation specimen as described above, and an object of the present invention is to prepare an observation specimen by fixing a thin slice sample on a slide glass. An object of the present invention is to provide an observation specimen preparation method that can remove wrinkles or curls generated in a thin slice sample at the time of thin cutting and obtain an observation specimen in a good state.
[0010]
[Means for Solving the Problems]
The preparation method of the observation specimen of the present invention is as follows:
Slice the surface layer of the sample block where the specimen is embedded in the embedding material,
The sliced sample cut out is adsorbed to the lower surface of the carrier tape by static electricity and removed.
Apply water or a water-based adhesive to the top surface of the slide glass,
The thin slice sample is brought into close contact with the slide glass, and the thin slice sample is transferred from the carrier tape onto the slide glass.
In the method for preparing an observation specimen in which a thin slice sample is fixed on the slide glass by heating the slide glass,
Before the thin slice sample is brought into close contact with the slide glass, the adhesive liquid is preliminarily heated to a temperature of 30 ° C. or higher and 50 ° C. or lower.
[0011]
According to the preparation method of the specimen for observation of the present invention, the adhesive liquid on the slide glass is heated to 30 ° C. or higher and 50 ° C. or lower in advance compared to the case where the adhesive liquid on the slide glass is about room temperature. Since the thin section is placed, wrinkles or curls generated in the thin section sample at the time of thin cutting are removed, and an observation specimen in a good state can be obtained.
[0012]
Preferably, after the thin slice sample is transferred onto the slide glass, the slide glass is heated to a temperature of 40 ° C. or higher and 50 ° C. or lower to fix the thin slice sample on the slide glass.
[0013]
Preferably, after heating the adhesive liquid to a temperature of 30 ° C. or higher and 50 ° C. or lower, the adhesive liquid is dropped or sprayed on the slide glass.
[0014]
Instead of this, the adhesive liquid may be heated to a temperature of 30 ° C. or higher and 50 ° C. or lower together with the slide glass before the thin slice sample is brought into close contact with the slide glass.
[0015]
The observation specimen preparation apparatus of the present invention used for carrying out the above method is:
A cutter for slicing the surface layer portion of the sample block in which the specimen is embedded in the embedding material,
A carrier tape supply mechanism for supplying the carrier tape above the surface of the sample block, and for collecting and collecting the cut sliced sample by electrostatic force on the lower surface of the carrier tape;
An adhesive liquid supply device that drops or sprays water or an adhesive liquid mainly composed of water on the upper surface of the slide glass;
A slide glass supply mechanism for supplying the slide glass to the lower side of the running trajectory of the carrier tape and receiving the thin slice sample from the carrier tape onto the slide glass;
A heating and fixing device for heating the slide glass on which the thin section sample is placed to a temperature of 40 ° C. or more and 50 ° C. or less and fixing the thin section sample on the slide glass;
In the preparation device of the specimen for observation in which the surface layer part of the sample block is sliced and the cut sliced sample is fixed on the slide glass and collected,
The adhesive liquid supply device includes a heating means for heating the adhesive liquid to a temperature of 30 ° C. or higher and 50 ° C. or lower before dropping or spraying the adhesive liquid.
[0016]
As described above, instead of providing a heating means in the adhesive liquid supply device, before receiving the thin slice sample from the carrier tape onto the slide glass, the adhesive liquid is 30 ° C. or higher on the slide glass. You may provide the heating means which heats to the temperature of 50 degrees C or less.
[0017]
DETAILED DESCRIPTION OF THE INVENTION
FIG. 1 and FIG. 2 show an outline of an apparatus used in the method for preparing an observation specimen according to the present invention. In the figure, 1 is a thin slice sample, 2 is a carrier tape, 3 is a slide glass, 4 is an adhesive liquid, 10 is a sample block, 23 is a cutter, 27 is a nozzle for supplying an adhesive liquid (adhesive liquid supply device), 29 is Represents a hot plate (heating fixing device).
[0018]
In this example, the sample block 10 is a biological sample embedded in paraffin. The carrier tape 2 is unwound from the unwinding roll 11 and is wound around the winding roll 12 through the guide rolls 13, 14, 15 and 16 in order. The traveling track of the carrier tape 2 is determined by these guide rolls.
[0019]
The carrier tape 2 travels in the horizontal direction in the section between the guide roll 13 and the guide roll 14 after being fed from the feed roll 11. A cutter 23 is disposed on the upstream side of the guide roll 14 adjacent to the guide roll 14 with a slight gap below the traveling track of the carrier tape 2.
[0020]
After the sample block 10 is positively charged by the first charging device 21, the sample block 10 is transferred to the front side of the guide roll 14 and set below the traveling track of the carrier tape 2. On the upper side of the running track of the carrier tape 2, a second charging device 22 is disposed above the sample block 10 with the carrier tape 2 interposed therebetween. When the second charging device 22 is started with the traveling of the carrier tape 2 stopped, the carrier tape 2 is negatively charged and the carrier tape 2 is adsorbed on the upper surface of the sample block 10. In this state, when the carrier tape 2 and the sample block 10 are sent out toward the cutter 23 in synchronization with each other, the surface layer portion of the sample block 10 is sliced by the cutter 23. The sliced sample 1 cut out is attracted to the lower surface of the carrier tape 2 by electrostatic force and collected.
[0021]
The thin slice sample 1 collected in this way is transferred to a section between the guide roll 15 and the guide roll 16 as the carrier tape 2 travels. In this section, as shown in FIG. 2, an adhesive liquid supply station 26 and a sample fixing station 28 are provided adjacent to both sides of the traveling track of the carrier tape 2, respectively. The adhesive liquid supply station 26 is provided with an adhesive liquid supply nozzle 27, and the sample fixing station 28 is provided with a hot plate 29. The slide glass 3 is sent from the adhesive liquid supply station 26 to the lower side of the traveling track of the carrier tape 2 by a conveying device (slide glass supply mechanism; not shown), and then from the lower side of the traveling track of the carrier tape 2. It is sent to the sample fixing station 28.
[0022]
After the thin slice sample 1 reaches the above-mentioned section, the carrier tape 2 is stopped once and the slide glass 3 is set below the thin slice sample 1. The adhesive liquid 4 is previously applied to the upper surface of the slide glass 3 by dropping the adhesive liquid 4 (water in this example) heated to 50 ° C. from the nozzle 27 at the adhesive liquid supply station 26. Has been.
[0023]
In this state, the guide roll 16 is pushed down to bring the thin slice sample 1 into close contact with the upper surface of the slide glass 3. Next, when the guide roll 16 is pulled upward, the thin slice sample 1 is adhered onto the slide glass 3 by the adhesive liquid and transferred to the slide glass 3 side.
[0024]
Next, the slide glass 3 on which the thin section sample 1 is placed is transferred to the sample fixing station 28 and placed on the hot plate 29. When the slide glass 3 is heated to 50 ° C. and held for about 30 minutes on the hot plate 29, the adhesive liquid evaporates and the thin slice sample 1 is fixed to the upper surface of the slide glass 3. In this way, an observation sample is created.
[0025]
FIG. 3 shows an example of an apparatus for previously heating the adhesive liquid to a predetermined temperature in the adhesive liquid supply station 26 (FIG. 2). In this example, a heater 32 is attached around the tank 31 in which the adhesive liquid 4 is stored, and the adhesive liquid 4 in the tank 31 is heated. The heated adhesive liquid 4 is dropped onto the slide glass 3 through a nozzle 34 provided at the bottom of the tank 31. A temperature measuring sensor 33 is immersed in the tank 31.
[0026]
FIG. 4 shows another example of an apparatus for preheating the adhesive liquid. In this example, a heater 32 is attached around the tank 31 in which the adhesive liquid 4 is stored, and the adhesive liquid 4 in the tank 31 is heated. The heated adhesive liquid 4 is sprayed onto the slide glass 3 through a spray nozzle 35 provided at the bottom of the tank 31.
[0027]
FIG. 5 shows another example of an apparatus for preheating the adhesive liquid. In this example, the slide glass 3 is placed on the hot plate 41 and the adhesive liquid 4 is heated. After the adhesive liquid 4 is heated to a predetermined temperature, the slide glass 3 is set below the traveling track of the carrier tape 2 (between the guide rolls 15 and 16 in FIG. 1).
[0028]
FIG. 6 shows another example of an apparatus for preheating the adhesive liquid. In this example, the slide glass 3 is placed on the support base 43, and the adhesive liquid 4 is heated from above using the infrared lamp 45. After the adhesive liquid 4 is heated to a predetermined temperature, the slide glass 3 is set below the traveling track of the carrier tape 2.
[0029]
【The invention's effect】
According to the method for preparing an observation specimen of the present invention, the adhesive liquid on the slide glass is preliminarily heated to a temperature of 30 ° C. or more and 50 ° C. or less, so that the wrinkles generated in the thin slice sample at the time of slicing Alternatively, since the curl is removed, the observation specimen can be prepared in a good state.
[Brief description of the drawings]
FIG. 1 is a diagram showing an outline of an apparatus for carrying out a method for producing an observation specimen according to the present invention.
FIG. 2 is a view of the device shown in FIG.
FIG. 3 is a view showing an example of an apparatus for previously heating an adhesive liquid.
FIG. 4 is a view showing another example of an apparatus for preheating the adhesive liquid.
FIG. 5 is a view showing another example of an apparatus for previously heating an adhesive liquid.
FIG. 6 is a view showing another example of an apparatus for previously heating the adhesive liquid.
FIGS. 7A and 7B are diagrams illustrating an outline of a conventional method for preparing an observation specimen, and FIGS. 7A to 7C are diagrams illustrating each process. FIGS.
[Explanation of symbols]
1. Thin section sample,
2 ... carrier tape,
3 ... slide glass,
4 ... Adhesive liquid,
11 ... unwinding roll 12 ... winding roll,
13, 14, 15, 16 ... guide rolls,
21 ... 1st charging device,
22 ... second charging device,
23 ... Cutter,
26: Adhesive liquid supply station,
27 ... Nozzle for supplying adhesive liquid,
28 ... Sample fixing station,
29 ... Hot plate,
31 ... Tank,
32 ... heater,
33 ... Sensor for temperature measurement,
34 ... Nozzle,
35 ... Spray nozzle,
41 ... Hot plate,
43 ... support stand,
45: Infrared lamp.

Claims (6)

Slice the surface layer of the sample block where the specimen is embedded in the embedding material,
The sliced sample cut out is adsorbed to the lower surface of the carrier tape by static electricity and removed.
Apply water or a water-based adhesive to the top surface of the slide glass,
The thin slice sample is brought into close contact with the slide glass, and the thin slice sample is transferred from the carrier tape onto the slide glass.
In the method for preparing an observation specimen in which a thin slice sample is fixed on the slide glass by heating the slide glass,
A method for preparing an observation specimen, wherein the adhesive liquid is heated to a temperature of 30 ° C. or higher and 50 ° C. or lower in advance before the thin slice sample is brought into close contact with the slide glass. The thin slice sample is fixed on the slide glass by transferring the thin slice sample onto the slide glass and then heating the slide glass to a temperature of 40 ° C or higher and 50 ° C or lower. Item 2. A method for producing an observation specimen according to Item 1. 2. The method for producing an observation specimen according to claim 1, wherein after the adhesive liquid is heated to a temperature of 30 ° C. or higher and 50 ° C. or lower, the adhesive liquid is dropped or sprayed on the slide glass. The specimen for observation according to claim 1, wherein the adhesive liquid is heated to a temperature of 30 ° C. or more and 50 ° C. or less together with the slide glass before the thin slice sample is brought into close contact with the slide glass. Manufacturing method. A cutter for slicing the surface layer portion of the sample block in which the specimen is embedded in the embedding material,
A carrier tape supply mechanism for supplying the carrier tape above the surface of the sample block, and for collecting and collecting the cut sliced sample by electrostatic force on the lower surface of the carrier tape;
An adhesive liquid supply device that drops or sprays water or an adhesive liquid mainly composed of water on the upper surface of the slide glass;
A slide glass supply mechanism for supplying the slide glass to the lower side of the running trajectory of the carrier tape and receiving the thin slice sample from the carrier tape onto the slide glass;
A heating and fixing device for heating the slide glass on which the thin section sample is placed to a temperature of 40 ° C. or more and 50 ° C. or less and fixing the thin section sample on the slide glass;
In the preparation device of the specimen for observation in which the surface layer part of the sample block is sliced and the cut sliced sample is fixed on the slide glass and collected,
The observation liquid preparation apparatus according to claim 1, further comprising a heating means for heating the adhesive liquid to a temperature of 30 ° C. or higher and 50 ° C. or lower before dropping or spraying the adhesive liquid. A cutter for slicing the surface layer portion of the sample block in which the specimen is embedded in the embedding material,
A carrier tape supply mechanism for supplying the carrier tape above the surface of the sample block, and for collecting and collecting the cut sliced sample by electrostatic force on the lower surface of the carrier tape;
An adhesive liquid supply device that drops or sprays water or an adhesive liquid mainly composed of water on the upper surface of the slide glass;
A slide glass supply mechanism for supplying the slide glass to the lower side of the running trajectory of the carrier tape and receiving the thin slice sample from the carrier tape onto the slide glass;
A heating and fixing device for heating the slide glass on which the thin section sample is placed to a temperature of 40 ° C. or more and 50 ° C. or less and fixing the thin section sample on the slide glass;
In the preparation device of the specimen for observation in which the surface layer part of the sample block is sliced and the cut sliced sample is fixed on the slide glass and collected,
Observation comprising a heating means for heating the adhesive liquid to a temperature of 30 ° C. or higher and 50 ° C. or lower on the slide glass before receiving the thin slice sample from the carrier tape onto the slide glass. Specimen preparation equipment.

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