JP2006010620A - Method of preparing cross-sectional specimen for microscopic observation and ultramicrotome - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a specimen preparing method suited to evaluate such a specimen as might degenerate due to resin embedding. <P>SOLUTION: This method of preparing a cross-sectional specimen for microscopic observation is characterized in that a specimen is not embedded in resin, etc. but directly fixed to a specimen arm when cutting off the specimen by using ultramicrotome, the specimen is cut off with a knife boat not filled with liquid such as water, and a cross section formed on the specimen block side by the cutting is used for microscopic observation. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an ultramicrotome for producing a cross section of a solid sample, and particularly to an ultramicrotome capable of producing a cross section of a sample by dry cutting without embedding a resin.

Conventionally, an ultramicrotome has been used as a means for preparing a thinned sample to be observed with an electron microscope or an optical microscope. The sliced section is prepared by embedding a sample in a resin or the like, and then fixing the sample to an ultramicrotome arm, moving the arm up and down, and cutting with a diamond or glass knife. The cut section will float on the water surface of the knife boat. This cut piece was scooped into a mesh and used as a microscope sample for a transmission electron microscope. Alternatively, a sample block cross section obtained by cutting has been used as a microscope sample such as an optical microscope, a laser microscope, an SPM, an AFM, and a scanning electron microscope.
Japanese Patent Application No. 8-136417

  However, when a fibrous sample such as paper is embedded with a resin, there is a problem that the sample swells due to the resin that has penetrated between the fibers and the structure is altered. Also, when using a laser microscope, SPM, AFM, etc. to evaluate irregularities such as paper, coated paper coating layers, or pigment color materials, the embedding resin fills the gaps between the paper fibers, coat layers, and pigment color materials. Therefore, unevenness information could not be obtained. The present invention was obtained as a result of intensive studies to solve the above problems, and provides a sample preparation method suitable for evaluation of samples that are altered by resin embedding.

  For this reason, in the present invention, the section of the cut sample block is observed instead of using the section generated by cutting as a microscope sample. In this case, the first feature of the present invention is that the sample is directly fixed to the sample arm without being embedded in the resin. Also, touching the sample with water causes deterioration, so use the knife boat without water. When cutting is performed in such a state, the sliced section is hardly suitable for observation due to chattering or the like. However, the remaining sample block is not altered by swelling or the like, and the original structure of the sample is maintained. In addition, the cross section obtained in this way is flat, and it has been clarified that the cross section is ideal for observation with an optical microscope, a laser microscope, a scanning probe microscope, a scanning electron microscope, or the like.

  Another feature of the present invention is that an ultramicrotome is provided with a blowing mechanism for a sample block or a knife. By this air blowing mechanism, it is possible to remove a section attached to the sample block or the knife when the sample is cut. As described above, when an ultramicrotome is used without putting water into the knife boat, the cut section may adhere to the knife blade or the sample, which may hinder smooth cutting or damage the cut surface. In some cases, the section attached to the sample surface obstructs observation. In order to prevent this, the present invention is characterized in that when the sample fixed to the sample arm moves to a position where it does not touch the knife, it blows air to the knife or the sample block and blows off the attached section. The vertical movement of the sample arm must be repeated until the cut surface reaches the position of the sample to be observed, and generally requires a considerable amount of time. For this reason, in order to blow air manually, an operator is restrained by the ultra microtome device during the cutting process, which is troublesome and inefficient. Therefore, in the present invention, in the process in which the arm to which the sample is fixed repeats the vertical movement, it is provided with a mechanism that automatically blows air when the sample moves to a position where it does not touch the knife in synchronization with the vertical movement of the arm. Features. Furthermore, the present invention provides the air blowing mechanism in which the air blowing direction can be freely selected from the knife side and the sample side fixed to the arm, or the air blowing direction can be changed alternately, the number of times the sample is cut, and the cutting time. Alternatively, the end of the cutting process of the sample can be set in advance by the product of one cutting thickness and the number of cuttings.

  In more detail, the present invention can solve the above problems by the following configuration.

  (1) When cutting a sample using an ultramicrotome, the sample is not directly embedded in a resin or the like, but directly fixed to a sample arm, and the knife boat is cut without filling a liquid such as water. A method for preparing a cross-sectional sample for microscopic observation, wherein the cross section on the side of the sample block formed by cutting is used for microscopic observation.

  (2) An ultramicrotome comprising a blower mechanism for the sample block or the knife as a means for preventing the sample section produced by the ultramicrotome from adhering to the sample block to be cut or the knife to be cut. .

  (3) The arm on which the sample is fixed repeats the vertical movement at the position of the knife, so that the sample fixed on the arm touches the knife in the process of cutting the sample in synchronization with the vertical movement of the arm. An ultramicrotome comprising a mechanism that automatically blows air to the knife or the sample block only when it is moved to a non-position.

  (4) In the air blowing mechanism, the air blowing direction can be freely selected from the knife side and the sample side fixed to the arm, or the air blowing direction can be changed alternately. Ultra microtome.

  (5) In the above (2) or (3), the end of the cutting process of the sample can be set in advance by the number of times of cutting of the sample, the cutting time, or the product of one cutting thickness and the number of times of cutting. The described ultramicrotome.

  According to the present invention, it is possible to efficiently produce a cross section of a sample that is suitable for microscopic observation without deterioration of the sample and without contamination due to cutting.

  Hereinafter, the best mode for carrying out the present invention will be described in detail with reference to examples.

  The blower mechanism may be anything such as compressed air, nitrogen gas, a fan using rotating blades, etc., but it is desirable that a strong wind force be obtained even in a moment to blow off the slice. In this embodiment, a small bomb filled with compressed nitrogen is used and air is blown by opening and closing a solenoid valve. The blower pressure is set by the pressure meter 7 so that a cut sample piece does not remain on the knife or the sample block. Adjusted to pressure.

  The present embodiment will be described with reference to FIG. Reference numeral 1 denotes an ultramicrotome body, and this apparatus can previously input a position where the sample arm 2 contacts the knife 5. Further, when the sample arm 2 is in the input position where it contacts the knife 5, the LED 12 is lit.

  In this embodiment, the mechanism is such that the electric signal that starts turning on the LED 12 is used as a trigger, and the electromagnetic valve 7 is opened for a certain period of time when the LED 12 is turned off to blow air. As shown in the figure, the air blowing port is branched from the gas nozzle support rod 9 in the two directions of the cut sample 10 and the knife 11. In addition, the piping from the branching portion to the tip portion has a bellows structure that can be freely deformed and adjusted in position, and the gas outlet is made thinner. Since the timer of the main body of the Ultrame can be set according to the number of times of cutting, the start / stop of the air blowing mechanism is also set according to the timer.

Schematic perspective view of the ultramicrotome body (gas cylinder set not attached). The schematic perspective view which shows a gas cylinder set attached to an ultra microtome. The schematic perspective view which shows the front-end | tip part vicinity of the ultramicrotome of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Ultramicrotome body 2 Sample arm 3 Metal sample chuck 4 Sample 5 Cutting knife 6 Nitrogen cylinder 7 Pressure meter 8 Solenoid valve 9 Gas nozzle support rod 10 Sample side bellows structure branch piping and gas outlet 11 Knob side bellows structure branch piping And gas outlet 12 LED
13 Cylinder open / close valve 14 Gas introduction pipe before branching

Claims (5)

  When cutting a sample using an ultramicrotome, the sample is not embedded in resin or the like, but directly fixed to the sample arm, and the knife boat is not filled with liquid such as water, and the sample is cut and formed by this cutting. A method for producing a cross-sectional sample for microscopic observation, wherein the cross section on the sample block side is used for microscopic observation.   An ultramicrotome comprising a blower mechanism for the sample block or the knife as a means for preventing a sample section produced by the ultramicrotome from adhering to a sample block to be cut or a knife to be cut.   The position where the sample fixed to the arm does not touch the knife in synchronization with the vertical movement of the arm in the process of cutting the sample by repeating the vertical movement of the arm to which the sample is fixed at the position of the knife An ultramicrotome comprising a mechanism that automatically blows air to the knife or the sample block only when moved to the position.   3. The ultramicrotome according to claim 2, wherein in the air blowing mechanism, two air blowing directions, a knife side and a sample side fixed to the arm, can be freely selected, or the air blowing direction can be changed alternately.   The ultramicrotome according to claim 2 or 3, wherein the end of the cutting process of the sample can be set in advance by the number of times of cutting the sample, the cutting time, or the product of the cutting thickness and the number of times of cutting.

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