USE OF A PLASTIC FILM IN THE FIELD OF LIGHT MICROSCOPY

Abstract

A plastic film used for the purpose of depositing and transporting of thin slices of samples in the field of light microscopy.

Claims

1. A plastic film used for the purpose of depositing and transporting of thin slices of samples in the field of light microscopy.

2. The plastic film according to claim 1 used for the purpose of depositing and transporting of thin slices of samples while cutting the slices with a microtome.

3. The plastic film according to claim 1 used for the purpose of depositing and transporting of samples while analysing the slices in a light microscope.

4. The plastic film according to claim 1 used for the purpose of depositing and transporting of samples while analysing the slices in a transmitted light microscope.

5. The plastic film according to claim 1, wherein the film is made of polypropylene.

6. The plastic film according to claim 1, wherein the film is made of an oriented polypropylene.

7. The plastic film according to claim 1, wherein the film is made of a biaxially oriented polypropylene.

8. The plastic film according to claim 1, wherein the film comprises at least two layers.

9. The plastic film according to claim 8, wherein the first layer is made of a biaxially oriented polypropylene and the second layer is made of a polypropylene.

10. The plastic film according to claim 8, wherein the film comprises three layers; wherein the first layer is made of a biaxially oriented polypropylene; and wherein the first layer is covered by a second layer and a third layer, wherein the second layer and the third layer are made of a polypropylene.

11. The plastic film according to claim 10, wherein the samples are placed on the free surface of the first layer.

12. The plastic film according to claim 1, wherein the film is transparent.

13. The plastic film according to claim 1, wherein the film is hydrophilically furnished.

14. The plastic film according to claim 13, wherein at least one surface is treated by corona treatment.

15. The plastic film according to claim 1, wherein the thickness of the film is between 20 m and 40 m.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Some embodiments of the microtome and the light microscope according to the invention are described in greater detail in the following. The drawings show in each case schematically:

[0039] FIG. 1 is a three-dimensional drawing of a microtome;

[0040] FIG. 2 shows the supply means in detail;

[0041] FIG. 3 shows the drive of the second holder in detail;

[0042] FIG. 4 is a front view of a sample supply means;

[0043] FIG. 5 is a plan view of a sample supply means;

[0044] FIG. 6 shows in detail the torque brake for the first holder;

[0045] FIG. 7 shows in detail an alternative torque brake.

DETAILED DESCRIPTION

[0046] FIG. 1 shows a microtome 1 suitable for preparing thin slices and ultrathin slices of samples having a thickness of approximately 1 m. In this context, the microtome 1 is in particular suitable for splitting up samples in the form of tissue samples which have previously been stabilised, for example, using epoxy resin.

[0047] The microtome 1 comprises a blade 2 in the form of a diamond knife for cutting thin slices from a sample, a sample holder 3 for supplying the sample to the blade 2, and a means 4 for receiving the thin slices, the means 4 being a film 5 on which the thin slices are deposited. In the present embodiment, the sample holder 3 along with the sample fastened thereon is moved in translation by means of an electric motor 16, the sample sliding along on the blade 2 and thin slices being cut off from the sample.

[0048] The film 5 is a continuous film which is unwound from a first spool 6 attached to a first holder 24 and wound up together with the thin slices on a second holder 7. The film 5 is a plastic film made of polypropylene. The film 5 comprises three layers. The first layer is made of a biaxially oriented polypropylene. The first layer is covered by a second layer and a third layer, the second layer and the third layer are made of a polypropylene. The thickness of the film 5 is 30 m. The film 5 is transparent and has a high light transmission.

[0049] The samples are placed on the free surface of the first layer. To enhance the adhesion of the samples, the film 5 is hydrophilically furnished. For this, the surface of the filmthe surface of the first layeris treated by corona treatment.

[0050] FIG. 2 shows in detail the supply means 8 disclosed in FIG. 1, which guides the film 5 to the diamond knife of the blade 2. In this context, the supply means 8 has a direction change 9 in the form of a high-grade steel rod. In the present case, the direction change 9 has a diameter of 1 mm. The supply means 8 further comprises a container 10 for receiving liquid, the film 5 changing direction in such a way that the film 5 is wetted with liquid before receiving thin slices. The blade 2 is likewise assigned to the container 10. After cutting, the thin slices are initially transported on the meniscus and finally removed from the film 5. In this context, both the thin slices and the film 5 are wetted with liquid. further direction change 21 causes the film 5 to change direction in such a way that the film 5 is guided via the container and wetted with liquid in the region of the direction change 9.

[0051] A heating means 25 for drying the thin slices deposited on the film 5 is assigned to the supply means 8. In an advantageous embodiment, the heating means 25 comprises a Peltier element, which is heated in parts by applying an electrical voltage. The thermal radiation produced as a result is sufficient to dry the thin slices and the film 5 prior to winding up. In this context, it is also advantageous that higher process speeds are possible.

[0052] In the method for preparing thin slices using a microtome 1, in a first step a sample is supplied to a blade 2 which cuts off thin slices from the sample, and in a second step the thin slices are deposited on a film 5 which is unwound from a first spool 6 and wound up together with the thin slices on a second holder 7.

[0053] The first spool 6 is provided with a torque brake 20 in the form of an eddy current brake (FIG. 7). Alternatively, the first spool can be provided with a torque brake 20 in the form of a disc filled with liquid (FIG. 6).

[0054] Further, it is conceivable to provide initial tension of the film by modification of the turning resistance, for example by clamping of the bearings.

[0055] FIG. 3 shows in detail the drive disclosed in FIG. 1, which winds the film 5 from the first spool 6 onto the second holder 7. The second holder 7 is operatively connected to a stepper motor 17. Both the first spool 6 and the second holder 7 are rotatably arranged on a shaft 18, the stepper motor 17 being connected to the second holder 7. The stepper motor 17 acts on the second holder 7 directly via a toothed belt drive 19. This ensures that the film 5 is wound onto the second holder 7 uniformly and continuously. A high-ratio transmission is assigned to the stepper motor 17. In the present embodiment, this ratio is 2,070:1.

[0056] In the present embodiment, the shaft 18 on which the second holder 7 is arranged is rigidly fastened in the supply means 8, and the second holder 7 is rotatably mounted on the shaft 18. In this embodiment, the toothed belt drive 19 is assigned to the second holder 7 and also rotatably mounted on the shaft 18.

[0057] In the present embodiment, the stepper motor 17 is mounted in the supply means 8 in such a way that said motor can be displaced parallel to the axis. Together with the stepper motor 17, the first holder 24 comprising the first spool 6 and the second holder 7 may also be displaced parallel to the axis. As a result, the position of the film 5 relative to the blade 2 can be displaced in such a way that the film 5 is always optimally positioned to receive the samples.

[0058] The arrangement comprising the first spool 6, the second holder 7, the supply means 8, the direction change 9, the stepper motor 17, the shaft 18, the torque brake 20 and the first holder 24 is rigidly connected to the microtome 1. Thus, the direction change 9 is firmly assigned to the blade 2. This ensures a constant distanceindependently from external influencesbetween the direction change 9, with the film 5 redirected on it, and the blade 2.

[0059] FIGS. 4 and 5 show a sample supply means 12 which can be arranged on the microscope slide of a light microscope. In this context, the sample supply means 12 is formed to supply the thin slices located on the wound-up film 5 on the second holder 7 to the microscope slide. In this context, the slices are supplied continuously.

[0060] The sample supply means 12 has a first retainer 13 for an empty spool 14 and a second retainer 15 for a second spool 23. The film 5 provided with the samples is located on the second spool 23. For this purpose, in an intermediate step, the film 5 may be wound up from the second holder 7 onto the second spool 23.

[0061] The second retainer 15 is provided with a metal disc 27 which is operatively connected to a permanent magnet 28. When the second spool 23 is rotating, the permanent magnet 28 induces an eddy current inside of the metal disc 27. Due to the ohmic resistance, part of the yielded energy is transformed into heat. Thus, the permanent magnet 28 and the metal disc 27 form a torque brake, more precisely an eddy current brake. The result is a resistance against rotation of the second spool 23 which then again results in an initial tension of the film 5 so that the film can be unwound uniformly. The distance between the permanent magnet 28 and metal disc 27 may be adjustable.

[0062] The empty spool 14 is operatively connected to a second electric motor 11 so as to wind the film 5 from the second spool 23 onto the empty spool 14 and thus to supply the thin slices to the microscope slide. The second electric motor 11 is provided with a high-ratio transmission. In the present case, the ratio is 2,070:1. The force transmission from the second electric motor 11 to the empty spool 14 takes place by way of a traction mechanism drive; in this case by means of a second toothed belt drive 26.

[0063] The sample supply means 12 is configured in such a way that the thin slices arranged on the film 5 can be supplied to the microscope slide continuously. For this purpose, the sample supply means 12 comprises a second electric motor 11 in the form of a stepper motor. The second electric motor 11 is operatively connected to the empty spool 14, and brings about continuous unwinding of the film 5 from the second spool 23. A second direction change 21 is arranged between the second spool 23 and the empty spool 14. The film 5 is guided via the second direction change 21, the second direction change 21 having a plateau 22, the analysing of the samples taking place on the plateau 22. The plateau is formed by a horizontally orientated support made of glass and ensures a constant and optimal working distance between objective of the light microscope and film.

[0064] In the method for analysing thin slices in a light microscope, a film 5provided with thin slices and wound-upis unwound and wound up on an empty spool 14 by means of a sample supply means 12, the thin slices deposited on the film 5 being supplied to the microscope slide during the unwinding from the second spool 23 and the winding onto the empty spool 14.

[0065] The spools 6, 14, 23 are formed as double-flanged spools.

[0066] FIG. 6 shows in detail the first spool 6 which is provided with a torque brake 20. In order for the film 5 also to be able to be unwound uniformly from the first spool 6, a torque brake 20, which is rotationally engaged with the first holder 24 or the first spool 6, is arranged on the shaft on which the first spool 6 is mounted.

[0067] In an advantageous embodiment, the torque brake 20 consists of a disc which is fastened to the shaft and provided with an annular cavity. The cavity is filled at least in part with a liquid, in the present case with a mixture of water and glycerol.

[0068] If the torque brake 20 is set in rotation together with the first spool 6, the disc moves, whilst the liquid remains in place. This results in a slight friction of the liquid on the inner wall of the cavity and a constant restoring force, leading to a constant slight resistance to the rotational movement. This resistance is sufficient for the film 5 to have a small bias and be unwound uniformly from the first spool 6. As a result of the liquid/solid tribological pairing, there is also no stick-slip effect, which would lead to a jerky movement of the film 5. As a result of the arrangement of the torque brake 20, a uniform tensile stress of the film 5 is always provided, leading to uniform unwinding and winding of the film 5. Such a torque brake 20 can be referred to as a gravity torque. Against the background of low torques needed for moving of the film, the torque brake is particularly advantageous as it is generating a low and consistent torque.

[0069] FIG. 7 depicts an alternative embodiment of a torque brake. In this embodiment, the first spool 6 is made of electrically conductive material. A magnet 28, here a permanent magnet 28, is assigned to the first spool 6, the permanent magnet 28 being attached adjustably on the microtome 1 relative to the first spool 6. The adjustment of the permanent magnet 28 is realized by a height adjustment 29 on which the permanent magnet 28 is arranged. When the first spool 6 is rotating, the permanent magnet 28 induces an eddy current inside of the electrically conductive sections of the first spool 6. Due to the ohmic resistance, part of the yielded energy is transformed into heat. Hence, the result is a resistance against rotation of the first spool 6 which then again results in an initial tension of the film 5. The closer the permanent magnet 28 is positioned on the first spool 6, the greater the resistance. In an alternative embodiment, the permanent magnet 28 can be formed as an electromagnet.