CA1171301A - Apparatus for intraoperative diagnosis - Google Patents
Apparatus for intraoperative diagnosisInfo
- Publication number
- CA1171301A CA1171301A CA000392552A CA392552A CA1171301A CA 1171301 A CA1171301 A CA 1171301A CA 000392552 A CA000392552 A CA 000392552A CA 392552 A CA392552 A CA 392552A CA 1171301 A CA1171301 A CA 1171301A
- Authority
- CA
- Canada
- Prior art keywords
- slide
- cryogenic liquid
- tissue
- chilled
- liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Abstract
Abstract:
An apparatus for histological analysis is described comprising a refrigerating device adapted to spray a cryogenic liquid on the surface of a microscope slide whereby the surface of the slide is chilled to at least -4°C. At least one cooling fan is provided which increases the evaporation of the cryogenic liquid and aids the chilling action. An automatic press plate is also provided which is adapted to apply a measurable and graduated pressure on an organ tissue section placed on the chilled slide, whereby a monolayer of the tissue exfoliates onto the slide. This has the advantage of providing a very thin monolayer of the tissue on the slide for analysis.
An apparatus for histological analysis is described comprising a refrigerating device adapted to spray a cryogenic liquid on the surface of a microscope slide whereby the surface of the slide is chilled to at least -4°C. At least one cooling fan is provided which increases the evaporation of the cryogenic liquid and aids the chilling action. An automatic press plate is also provided which is adapted to apply a measurable and graduated pressure on an organ tissue section placed on the chilled slide, whereby a monolayer of the tissue exfoliates onto the slide. This has the advantage of providing a very thin monolayer of the tissue on the slide for analysis.
Description
This invention relates to a process and apparatus for histological analysis. In particular, it relates to a method of preparing a sample for analysis by means of freeze imprinting.
The usual histological analysis is based on the use of a freezing cryotome. In medical practice, the surgeon takes a section of the tissue from the diseased part of the patient and gives this section to the pathologist who first freezes the section of the tissue and then cuts it into slices, applies color and carries out in this manner the microscopic diagnosis.
-~ Another method, which is very inaccurate, consists of pressing the tissue sample onto a slide whereb~ a very small amoun~ of the tissue material is left on the slide.
Unfortunately, this tissue is frequently in poor condition~
It is known, for Instance from DOS 2,704,300 that a possible techni~ue is to provide a small cylinder of anatomic material which, mounted in a suitable apparatus, is sliced with a blade such as to expose a fresh surface of the cylinder. Immediately after the cut, the surface ; is "fixed" by immersion of the cylinder in a refrigerating liquid, such as liquid nitrogen. The sample obtained in this manner is not only very thick, but the test surface suffers the usual damages caused by the friction produced by the blade.
:: ~
.
7~3~
Other apparatuses are also known for cooling anatomic samples, such as British Patent 1,023,597, or to cool a sample on a microscopic slide, such as French Patent 1,403,270, or to cool the slide body of a microscope as in U.S. Patent 3,580,658.
The apparatus of the present invention has very sub-stantial advantages over the previously known methods and, most importantly, it makes possible histological analysis of a monolayer.
The microscopic reproduction of a normal or a patho-logical organ is the most trustworthy and accurate method of analysis and the thinner is the layer of cells being examined under the microscope, the more trustworthy and accurate are the results. The ideal condition is to examine a monocellular layer which reproduces the structure exactly. In this manner it is possible to see in the positive picture the cells of the parenchyma, i.e., the epithelial and other tissues which perform the special functions of the organ and in the negative picture vessels and connective tissue, and stroma which will form some pockets. These pockets may also be interpreted quantitatively.
In accordance with the concept of the present inven-tion, a thermal shock is created between the section of the organ tissue and the slide on which it is being supported in such manner that as a result of the thermal shock, the cells are induced to exfoliate to form a monolayer on the slide. This is achieved while keeping intact the submicroscopic arrangement, and particularly the enzymatic liposomial arrangement.
Thus the present invention in its broadest aspect relates to an apparatus for histological analysis which comprises a refrigerating device adapted to spray a cryogenic liquid on the surface of a microscope slide whereby the surface of the slide is chilled to at least -4C, at least one cooling fan which increases the 3~
evaporation of the cryogenic liquid; and an automatic press plate adapted to apply a measurable and graduated pressure on an organ tissue section placed on the chilled ` slide, whereby a ~onolayer of the tissue exfoliates onto the slide.
In order to better illustrate the present invention, reference is made to the sensation of cold which one receives when one touches with bare hands a frozen surface, and particularly a very cold metal surface. Thus, it is well known that there is a sensation of instantaneous adhesiveness between the skin and the metal surface, these surfaces being at very different temperatures. In reality, if the metal surface is sufficiently cold, the skin when removed from the surface leaves thereon a thin cellular layer.
With the apparatus of this invention, the press plate may also be temperature controlled, e.g. chilled to a desired temperature. It is also possible to automatically advance the slide whereby several organ sample depositions may be made on the same slide. According to another pre-ferred feature, the slide may be vibrated by means of a vibrator to assist in the exfoliation of the sample.
Certain preferred embodiments of the invention will now be illustrated by the attached drawing which schematically illustrates the apparatus of the invention.
~; As shown, the device includes a cylinder 1 containing a cryogenic liquid 2, e.g. ethyl chloride. Above the liquid in the cylinder 1 is a gas phase 3 pressurized above the level of the cryogenic liquid 2.
The cylinder 1 connects via valve 4 and connector tube 5 to a nozzle 6 which is arranged to direct a spray or jet of cryogenic liquid onto the surface of a microscope slide 9. Fans 7 and 8 blow air, assisted by deflectors 7' and 8', on the lower surface of slide 9 to accelerate the evap-oration of the cryogenic liquid. This aids in speeding the cooling of the slide.
7~3~
The slide is supported by guides 10 and 11 which permit lateral movement. The anatomic section to be exfoliated is placed on the slide 9 and is compressed on it by means of the press plate 12, which is connected to a toothed shaft 13, moved up and down by a toothed wheel 14 forming part of a pinion gear arrangement. With this arrangement, the press plate 12 may exert a graduated pressure on the section bein~ analyzed. The anatomic section is then pressed onto the slide 9 after the latter has reached a fixed temperature. This usually occurs 3 to 4 minutes after the valve 4 has been opened and the cooling fans 7 and 8 have been operational.
FinaIly, the guides 10 and 11 and the slide 9 are subjected to vibration by means of a vibrator 15 and this subjects the anatomic section to a mechanical shock, which in conjunction with the thermal shock facilitates the exfoliation.
The apparatus may also be provided with a device for automatically advancing the slide 9 so that several depositions at different pressures may be made on the same slide and information may be obtained with respect to the degree of separation of the cells after the second, third, fourth, etc. pressing. In this manner, it is possible to reconstruct, layer by layer, the properties of the tissue.
Such results could not be achieved by MeanS of conventional histological analysis. The degree of cellular adhesion to the chilled slide is also an indication of the density of the neoplasiae.
It will also be understood that various modifications of the present invention are possible within the scope and spirit of the present claims. For instance, the fans 7 and 8 and vibrating device 15 may be electrically activated by a battery or by an electric current. The cryogenic liquid may be varied depending on the desired results and there may be used a gaseous material which can remain liquid at very low temperatures, such as li~uid nitrogen.
The pressurized gas may of the same material as the cryo-genic liquid and the rate of discharge of the liquid when the valve 4 is open may be accelerated by warming the cyl-inder particularly in the region of the pressurized gas 3.
The apparatus of the present invention has the very important advantage of very small dimensions and may have a size of at the most in the range of 30 to 40 cm. This is a feature which makes it extremely easy to handle and easy to transport.
The cooling time varies from one bioptic section to another and also depends upon the thickness of the slide, but in any event, the period of time is very short, ranging to a few tenths of a second. At the end of the cooling period, the section is removed with tweezers, the slide is removed from the guide and the slide is then ready for instantaneous application of color.
According to another embodiment, the slide may be precolored, in which case the system requires no more than 3 to 5 minutes total time Erom when the apparatus begins to be operated up to the time when the slide is ready for the diagnostic analysis.
From experiments which have been carried out with the apparatus of the present invention it has been determined that:
1. information can be obtained on the structure of the parenchima being examined which undergoes exfoliation in the form of a monolayer on the slide;
The usual histological analysis is based on the use of a freezing cryotome. In medical practice, the surgeon takes a section of the tissue from the diseased part of the patient and gives this section to the pathologist who first freezes the section of the tissue and then cuts it into slices, applies color and carries out in this manner the microscopic diagnosis.
-~ Another method, which is very inaccurate, consists of pressing the tissue sample onto a slide whereb~ a very small amoun~ of the tissue material is left on the slide.
Unfortunately, this tissue is frequently in poor condition~
It is known, for Instance from DOS 2,704,300 that a possible techni~ue is to provide a small cylinder of anatomic material which, mounted in a suitable apparatus, is sliced with a blade such as to expose a fresh surface of the cylinder. Immediately after the cut, the surface ; is "fixed" by immersion of the cylinder in a refrigerating liquid, such as liquid nitrogen. The sample obtained in this manner is not only very thick, but the test surface suffers the usual damages caused by the friction produced by the blade.
:: ~
.
7~3~
Other apparatuses are also known for cooling anatomic samples, such as British Patent 1,023,597, or to cool a sample on a microscopic slide, such as French Patent 1,403,270, or to cool the slide body of a microscope as in U.S. Patent 3,580,658.
The apparatus of the present invention has very sub-stantial advantages over the previously known methods and, most importantly, it makes possible histological analysis of a monolayer.
The microscopic reproduction of a normal or a patho-logical organ is the most trustworthy and accurate method of analysis and the thinner is the layer of cells being examined under the microscope, the more trustworthy and accurate are the results. The ideal condition is to examine a monocellular layer which reproduces the structure exactly. In this manner it is possible to see in the positive picture the cells of the parenchyma, i.e., the epithelial and other tissues which perform the special functions of the organ and in the negative picture vessels and connective tissue, and stroma which will form some pockets. These pockets may also be interpreted quantitatively.
In accordance with the concept of the present inven-tion, a thermal shock is created between the section of the organ tissue and the slide on which it is being supported in such manner that as a result of the thermal shock, the cells are induced to exfoliate to form a monolayer on the slide. This is achieved while keeping intact the submicroscopic arrangement, and particularly the enzymatic liposomial arrangement.
Thus the present invention in its broadest aspect relates to an apparatus for histological analysis which comprises a refrigerating device adapted to spray a cryogenic liquid on the surface of a microscope slide whereby the surface of the slide is chilled to at least -4C, at least one cooling fan which increases the 3~
evaporation of the cryogenic liquid; and an automatic press plate adapted to apply a measurable and graduated pressure on an organ tissue section placed on the chilled ` slide, whereby a ~onolayer of the tissue exfoliates onto the slide.
In order to better illustrate the present invention, reference is made to the sensation of cold which one receives when one touches with bare hands a frozen surface, and particularly a very cold metal surface. Thus, it is well known that there is a sensation of instantaneous adhesiveness between the skin and the metal surface, these surfaces being at very different temperatures. In reality, if the metal surface is sufficiently cold, the skin when removed from the surface leaves thereon a thin cellular layer.
With the apparatus of this invention, the press plate may also be temperature controlled, e.g. chilled to a desired temperature. It is also possible to automatically advance the slide whereby several organ sample depositions may be made on the same slide. According to another pre-ferred feature, the slide may be vibrated by means of a vibrator to assist in the exfoliation of the sample.
Certain preferred embodiments of the invention will now be illustrated by the attached drawing which schematically illustrates the apparatus of the invention.
~; As shown, the device includes a cylinder 1 containing a cryogenic liquid 2, e.g. ethyl chloride. Above the liquid in the cylinder 1 is a gas phase 3 pressurized above the level of the cryogenic liquid 2.
The cylinder 1 connects via valve 4 and connector tube 5 to a nozzle 6 which is arranged to direct a spray or jet of cryogenic liquid onto the surface of a microscope slide 9. Fans 7 and 8 blow air, assisted by deflectors 7' and 8', on the lower surface of slide 9 to accelerate the evap-oration of the cryogenic liquid. This aids in speeding the cooling of the slide.
7~3~
The slide is supported by guides 10 and 11 which permit lateral movement. The anatomic section to be exfoliated is placed on the slide 9 and is compressed on it by means of the press plate 12, which is connected to a toothed shaft 13, moved up and down by a toothed wheel 14 forming part of a pinion gear arrangement. With this arrangement, the press plate 12 may exert a graduated pressure on the section bein~ analyzed. The anatomic section is then pressed onto the slide 9 after the latter has reached a fixed temperature. This usually occurs 3 to 4 minutes after the valve 4 has been opened and the cooling fans 7 and 8 have been operational.
FinaIly, the guides 10 and 11 and the slide 9 are subjected to vibration by means of a vibrator 15 and this subjects the anatomic section to a mechanical shock, which in conjunction with the thermal shock facilitates the exfoliation.
The apparatus may also be provided with a device for automatically advancing the slide 9 so that several depositions at different pressures may be made on the same slide and information may be obtained with respect to the degree of separation of the cells after the second, third, fourth, etc. pressing. In this manner, it is possible to reconstruct, layer by layer, the properties of the tissue.
Such results could not be achieved by MeanS of conventional histological analysis. The degree of cellular adhesion to the chilled slide is also an indication of the density of the neoplasiae.
It will also be understood that various modifications of the present invention are possible within the scope and spirit of the present claims. For instance, the fans 7 and 8 and vibrating device 15 may be electrically activated by a battery or by an electric current. The cryogenic liquid may be varied depending on the desired results and there may be used a gaseous material which can remain liquid at very low temperatures, such as li~uid nitrogen.
The pressurized gas may of the same material as the cryo-genic liquid and the rate of discharge of the liquid when the valve 4 is open may be accelerated by warming the cyl-inder particularly in the region of the pressurized gas 3.
The apparatus of the present invention has the very important advantage of very small dimensions and may have a size of at the most in the range of 30 to 40 cm. This is a feature which makes it extremely easy to handle and easy to transport.
The cooling time varies from one bioptic section to another and also depends upon the thickness of the slide, but in any event, the period of time is very short, ranging to a few tenths of a second. At the end of the cooling period, the section is removed with tweezers, the slide is removed from the guide and the slide is then ready for instantaneous application of color.
According to another embodiment, the slide may be precolored, in which case the system requires no more than 3 to 5 minutes total time Erom when the apparatus begins to be operated up to the time when the slide is ready for the diagnostic analysis.
From experiments which have been carried out with the apparatus of the present invention it has been determined that:
1. information can be obtained on the structure of the parenchima being examined which undergoes exfoliation in the form of a monolayer on the slide;
2. the properties of the parenchima can be determined with respect to the density of the cells which have undergone desquamation;
3. information is available as to the presence in the negative of vascular structures,
4. it is also possible to determine the density of the cells in successive pressings as an index of the excess of cellular layers with respect to the possibility of the stroma being bound~
7~ 3~
Finally, the device of the present invention makes it possible to achieve a very substantial detailed diagnostic study of the cells with respect to nucleus, nucleoli, mitosis, the presence of mast cells and their quantitiz-ation ratio between cells and cells; in case of tumors, the relationship between neoplastic cells and normal cells;
relationship between tissues which are immuno-reactive and normal tissues or pathological tissues as in the case of infiltration in the liver of inflammatory substances or phlogistic reactions in areas of tumors to be examined in detail.
7~ 3~
Finally, the device of the present invention makes it possible to achieve a very substantial detailed diagnostic study of the cells with respect to nucleus, nucleoli, mitosis, the presence of mast cells and their quantitiz-ation ratio between cells and cells; in case of tumors, the relationship between neoplastic cells and normal cells;
relationship between tissues which are immuno-reactive and normal tissues or pathological tissues as in the case of infiltration in the liver of inflammatory substances or phlogistic reactions in areas of tumors to be examined in detail.
Claims (11)
1. An apparatus for histological analysis which comprises a refrigerating device adapted to spray a cryogenic liquid on the surface of a microscope slide whereby the surface of the slide is chilled to at least -4°C, at least one cooling fan which increases the evaporation of the cryo-genic liquid; and an automatic press plate adapted to apply a measurable and graduated pressure on an organ tissue section placed on the chilled slide, whereby a monolayer of the tissue exfoliates onto the slide.
2. An apparatus according to claim 1, including means for controlling the temperature of the press plate.
3. An apparatus according to claim 1, including means for automatically advancing the slide, whereby several organ sample depositions may be made on the same slide.
4. An apparatus according to claim 2, including means for automatically varying the pressure of the press plate during the advance of the slide.
5. An apparatus according to claim 1, wherein the refrigerating device comprises a cylinder containing cryogenic liquid.
6. An apparatus according to claim 1 wherein the refrigerating device includes a nozzle adapted to spray the cryogenic liquid onto a central portion of the slide.
7. An apparatus according to claim 1 including vibrator means for vibrating said slide.
8. A method of histological analysis which comprises directing a spray of cryogenic liquid on the surface of a microscope slide to chill said slide to at least -4°C, bringing a section of organ tissue to be analysed into contact with said chilled slide surface, pressing said tissue section against said chilled surface and subjecting the tissue to thermal shock whereby it exfoliates onto said slide.
9. A method according to claim 8 wherein the slide with the section of organ tissue being pressed thereon is subjected to mechanical vibration.
10. A method according to claim 8 wherein the cryogenic liquid is ethyl chloride.
11. A method according to claim 8 wherein the cryogenic liquid is liquid nitrogen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA000392552A CA1171301A (en) | 1981-12-17 | 1981-12-17 | Apparatus for intraoperative diagnosis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA000392552A CA1171301A (en) | 1981-12-17 | 1981-12-17 | Apparatus for intraoperative diagnosis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1171301A true CA1171301A (en) | 1984-07-24 |
Family
ID=4121640
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000392552A Expired CA1171301A (en) | 1981-12-17 | 1981-12-17 | Apparatus for intraoperative diagnosis |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA1171301A (en) |
-
1981
- 1981-12-17 CA CA000392552A patent/CA1171301A/en not_active Expired
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKEC | Expiry (correction) | ||
| MKEX | Expiry |