EP0570418A1 - A method of detecting the presence of anomalies in exfoliated cells using infrared spectroscopy - Google Patents

A method of detecting the presence of anomalies in exfoliated cells using infrared spectroscopy

Info

Publication number
EP0570418A1
EP0570418A1 EP92903692A EP92903692A EP0570418A1 EP 0570418 A1 EP0570418 A1 EP 0570418A1 EP 92903692 A EP92903692 A EP 92903692A EP 92903692 A EP92903692 A EP 92903692A EP 0570418 A1 EP0570418 A1 EP 0570418A1
Authority
EP
European Patent Office
Prior art keywords
specimen
exfoliated
cells
infrared
tissue
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.)
Withdrawn
Application number
EP92903692A
Other languages
German (de)
French (fr)
Inventor
Patrick T. T. Wong
Basil Rigas
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
National Research Council of Canada
Original Assignee
National Research Council of Canada
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by National Research Council of Canada filed Critical National Research Council of Canada
Publication of EP0570418A1 publication Critical patent/EP0570418A1/en
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light

Definitions

  • Exfoliated tissue or cell specimens may also be obtained from, for example, the trachea and bronchi, stomach, rectum and colon, urinary tract, serous sac fluids, cyst fluids, synovial fluids, glandular secretions and exudates. Methods for obtaining, collecting and, if necessary, preserving the specimens may vary according to the preference.
  • a method of detecting the presence of anomalies in exfoliated cells using infrared spectroscopy comprising: a) directing a beam of infrared light at a specimen of exfoliated cells, and b) determining if an anomaly is present in the specimen by spectral analysis of the infrared absorption of the specimen, at at least one range of frequencies, to ascertain whether at least one change in the infrared absorption characteristics has occurred, due to vibration of at least one functional group of molecules, in the specimen, which is characteristic of that anomaly.
  • the said at least one change in the infrared absorption characteristics may be a change in absorption intensity at a particular frequency, a change of frequency at which a particular absorption occurs, or a different pressure applied to the functional group causing a change of frequency at which a particular absorption occurs.
  • the functional group of molecules may be in at least one of the following molecules, carbohydrates, nucleic acids, tissue proteins or membrane lipids.
  • the functional group may be a phosphodiester group in nucleic acids, a C-OH group in tissue protein and carbohydrates, or a CH2 group in lipids.
  • the functional group may be at least one functional group selected from the groups consisting of carbohydrates, phosphodiester groups in nucleic acids, COH groups of tissue proteins or carbohydrates and CH ? group of lipids.
  • the specimen may be a Papanicolau smear.
  • the specimen may be a cervical specimen.
  • the specimen may be an endocervical specimen.
  • the specimen may be an ectocervical specimen.
  • the specimen may be a vaginal specimen.
  • the specimen may be a uterus specimen.
  • Figure 1 is a block diagram of an apparatus for detecting the presence of anomalies in exfoliated cells using infrared spectroscopy
  • Figure 2 shows infrared spectra in the frequency range 950 to 1,350 cm , obtained from normal healthy, exfoliated cervical cell specimens designated and from exfoliated cervical cell specimens designated and diagnosed in a conventional, cytological manner as malignant,
  • Figures 3 to 5 show the frequency shifts of the infrared bands at ⁇ 1082 cm -1 , and ⁇ 1155 cm -1 , and the changes in the intensity ratios between the infrared bands at ⁇ 1025 cm -1 and ⁇ 1082 cm -1 , respectively, between healthy, exfoliated cervical cell specimens and those diagnosed in a conventional, cytological manner as malignant,
  • Figure 6 shows the infrared spectra depicted in Figure 2, for the bands at "-1240 cm , enlarged and superimposed,
  • Figure 8 shows a comparison of the pressure dependencies of the vibrational frequencies of PO2 groups of nucleic acids in exfoliated cells of cervical tissues
  • Figure 10 shows the corresponding third power derivative spectra of the infrared spectra shown in Figure 9
  • Figure 11 shows a comparison of the pressure dependancies of the frequencies of the component bands of the C-0 stretching modes in the frequency range 1140 cm -1 to 1190 cm "1 between normal, healthy, exfoliated cervical tissue specimens and those diagnosed in a conventional, cytological manner as maligna n t,
  • infrared (IR) spectra were obtained as described with reference to Fig. 1 from exfoliated cervical cell specimens, that were collected from 25 females undergoing a surveillance Papanicolaou test (cervical cytology) following a standard "scraping" or “brushing” procedure. These specimens consisted of epithelial cells, inflammatory cells, red blood cells and bacteria that are part of the normal flora.
  • the most prominent differences include the following findings in malignant specimens: a) severe change in the intensity of the bands at 1025 cm -1 , 1047 cm -1 (it appears as a shoulder on'- 1025 cm “1 band), 1082 cm “1 , - 1155 cm -1 , -1244 cm “1 and -1303 cm “1 , b) significant shift of ⁇ 1082 cm “1 , » 1155 cm “1 and —1244 cm” 1 peaks, and c) a new band peaking at «_ 970 cm -1 .
  • the bands at ⁇ 1025 cm -1 and ⁇ 1047 cm “1 were found to be due to carbohydrates, and glycogen was found to make an overwhelming contribution to the intensity of these bands.
  • the bands at «- 1082 cm “1 and rm 1244 cm “1 were found to be due to the symmetric and asymmetric phosphate (POj) stretching modes, respectively. They were found to originate mainly in the vibrations of the phosphodiester groups of cellular nucleic acids.
  • the band at r*> H55 cm " was found to be due to the C-0 stretching mode of the C-OH groups of, among others, the residues of the amino acids serine, threonine and tyrosine in cell proteins as well as the C-OH groups of carbohydrates.
  • FIG. 6 is an enlargement of a portion of Figure 2, and the corresponding third power derivative spectra shown in Figure 7, a more detailed study of the asymmetric phosphate stretching band revealed that in malignant, exfoliated cervical tissues, shown , it consisted of two overlapping bands, one of which, the lower frequency band, was markedly increased when compared to normal healthy, exfoliated cells.
  • the frequency of the asymmetric PO stretching band was known at the time that the tests were made to be -*- 1220 cm "1 , when the P ⁇ 2 ⁇ group was fully hydrogen-bonded, and greater than 1240 cm "1 when it is not hydrogen-bonded.
  • the prominent differences for dysplasia specimens as compared to normal specimens include: a) severe change in the intensity of the bands at 1025 cm “1 , ⁇ * 1082 cm -1 , ⁇ -- 1244 cm “1 and 1303 cm “1 , and b) a significant shift in the frequency of the bands at ⁇ • ⁇ - 1047 cm -1 and ⁇ 1082 cm -1 .
  • the prominent differences for malignant specimens as compared to normal and dysplasia specimens include: a) severe change in the intensity of the bands at 970 cm “1 , - ⁇ 1025 cm “1 , ⁇ 1047 cm “1 , ⁇ 1082 cm “1 , -- 1155 cm “1 , 1244 cm “1 and r ⁇ * 1303 cm “1 , and b) a significant shift in the frequency of the bands at -*• 1025 cm “1 , -_- 1047 cm -1 ,
  • the prominent differences for the malignant specimens as compared to normal specimens include: a) a severe change in the intensity of the bands at ⁇ 1025 cm “1 , _ » 1154 cm “1 ,
  • the present invention is useful for, for example, detecting anomalies in exfoliated tissue or cell specimens from secretions, exudates, transudates of various human or other mammalian organs or tissues, and is particularly useful for detecting exfoliated cancerous tissues or cells from Papanicolaou cervical smears, i.e., endocervical smears, ectocervical smears, vaginal smears and uterus smears.
  • Exfoliated cell anomalies which may be detected according to the present invention include, for example, viruses, bacteria, fungi and other infections and non-infectious diseases.

Landscapes

  • Physics & Mathematics (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Dans ce procédé, on détecte la présence d'anomalies dans des cellules exfoliées ( par exemple dans un frottis du col utérin) à l'aide d'un spectroscope à infrarouge. On dirige un faisceau de lumière infrarouge sur des échantillons renfermant des cellules qui peuvent être des sécrétions, des exsudats, des transsudats, des raclages, des brossages ou des populations de cellules exfoliées de divers organes ou tissus obtenues d'une autre manière, et on détecte l'anomalie au niveau d'au moins une gamme de fréquences en déterminant s'il s'est produit une modification de l'absorption de la lumière infrarouge qui est due à la vibration d'un groupe fonctionnel dans, par exemple, des groupes photodiester d'acides nucléiques, des groupes COH de protéines tissulaires, de glucides, ou qui peut être due à des dispositions particulières de molécules lipidiques ou à des structures lipidiques anormales, présentes dans l'échantillon.In this method, the presence of abnormalities in exfoliated cells (for example in a cervical smear) is detected using an infrared spectroscope. A beam of infrared light is directed at samples containing cells which may be secretions, exudates, transsudates, scrapings, brushings or populations of cells exfoliated from various organs or tissues obtained in other ways, and detects the anomaly at at least one frequency range by determining whether there has been a change in the absorption of infrared light which is due to the vibration of a functional group in, for example, photodiester groups of nucleic acids, COH groups of tissue proteins, carbohydrates, or which may be due to specific arrangements of lipid molecules or abnormal lipid structures present in the sample.

Description

TITLE
A METHOD OF DETECTING THE PRESENCE OF ANOMALIES IN EXFOLIATED CELLS USING INFRARED SPECTROSCOPY
Technical Field
This invention relates to a method of detecting the presence of anomalies in exfoliated cells using infrared spectroscop .
Background Art
It is well known that examination for characteristic neoplastic cells in exfoliated tissue or cell specimens from, for example, the secretions, exudates, transudates, scrapings or brushings from various human organs or tissues may permit the diagnosis of malignancy. Exfoliative cytology has been found to be so valuable in the detection of early cancer of the uterus and uterine cervix that it is usually performed as a routine part of the annual examination for all adult women, see Pathologic Examination, Enfoliative Cytologic Study, Chapter 24, pages 362-365.
Exfoliated tissue or cell specimens may also be obtained from, for example, the trachea and bronchi, stomach, rectum and colon, urinary tract, serous sac fluids, cyst fluids, synovial fluids, glandular secretions and exudates. Methods for obtaining, collecting and, if necessary, preserving the specimens may vary according to the preference.
Each collected specimen is spread intact as an even layer over a clean microscope slide and submerged in, for example, an alcohol-ether fixative for about one hour. After being fixed, and without being allowed to dry, the layer is coated with glycerin and the whole sandwiched between two glass slides. The slides are then sent to a laboratory for staining and microscopic diagnosis by trained and experienced pathologists or cytologists.
While the microscopic diagnosis of exfoliated tissue or cell specimens is useful, the diagnostic interpretation: i) has to be carried out by skilled personnel, ii) is not completely reliable due, for example, to abundant mucous, trichomonas infection or atrophy, and iii) cannot accurately detect, for example, malignancy in tissues or cells in the pre-malignant or early malignant stages. There is a need for a method of detecting the presence of anomalies in exfoliated cells wherein: i) a simple test on the specimen, and the interpretation thereof, can be carried out by personnel having no medical skills and after a relatively brief course of training, ii) with proper care, interpretation of the test results is reliable, and iii) malignancy in tissues or cells can be detected in the pre-malignant or early malignant stages. It has already been proposed in the applicants ' co-pending Canadian Patent Application No. 2,008,831-1, filed January 29, 1990, "Method of Detecting the Presence of Anomalies in Biological Tissues and Cells in Natural or Cultured Form by Infrared Spectroscopy", P.T.T. Wong and B. Rigas, to detect the presence of anomalies in biological tissues and cells in natural and cultured form (e.g. cancerous tissues or cells) by infrared spectroscopy. A beam of infrared light is directed at a sample of tissue or cells in natural or cultured form containing the cells to be tested, and the anomaly is detected at at least one range of frequencies by determining whether changes in infrared absorption have occurred due to the vibration of at least one functional group of molecules present in the sample which is characteristic of the anomaly.
The method taught by P.T.T. Wong and B. Rigas is applied in the examples to biopsed tissue or cultured cells. In particular, P.T.T. Wong and B. Rigas, page 7 , line 20 to page 8, line 36, teach the detection of colorectal cancer from colonic ucosa immediately following surgical removal of a section or segment of the bowel. Disclosure of Invention
It has now been found, according to the present invention, that detection of the presence of anomalies in exfoliated cells by spectral analysis of the infrared absorption characteristics of the cells is possible. Mucin, water, inorganic salts, epithelial cells, red blood cells and leukocytes of the fluid do not give rise to any spurious interpretation of the infrared spectral absorption characteristics of the cells. According to the present invention there is provided a method of detecting the presence of anomalies in exfoliated cells using infrared spectroscopy, comprising: a) directing a beam of infrared light at a specimen of exfoliated cells, and b) determining if an anomaly is present in the specimen by spectral analysis of the infrared absorption of the specimen, at at least one range of frequencies, to ascertain whether at least one change in the infrared absorption characteristics has occurred, due to vibration of at least one functional group of molecules, in the specimen, which is characteristic of that anomaly. The said at least one change in the infrared absorption characteristics may be a change in absorption intensity at a particular frequency, a change of frequency at which a particular absorption occurs, or a different pressure applied to the functional group causing a change of frequency at which a particular absorption occurs.
The functional group of molecules may be in at least one of the following molecules, carbohydrates, nucleic acids, tissue proteins or membrane lipids.
The functional group may be a phosphodiester group in nucleic acids, a C-OH group in tissue protein and carbohydrates, or a CH2 group in lipids.
The functional group may be at least one functional group selected from the groups consisting of carbohydrates, phosphodiester groups in nucleic acids, COH groups of tissue proteins or carbohydrates and CH? group of lipids.
The specimen may be a Papanicolau smear. The specimen may be a cervical specimen. The specimen may be an endocervical specimen. The specimen may be an ectocervical specimen.
The specimen may be a vaginal specimen. The specimen may be a uterus specimen.
Brief Description of Drawings
In the accompanying drawings which illustrate, by way of example, embodiments of the present invention,
Figure 1 is a block diagram of an apparatus for detecting the presence of anomalies in exfoliated cells using infrared spectroscopy,
Figure 2 shows infrared spectra in the frequency range 950 to 1,350 cm , obtained from normal healthy, exfoliated cervical cell specimens designated and from exfoliated cervical cell specimens designated and diagnosed in a conventional, cytological manner as malignant,
Figures 3 to 5 show the frequency shifts of the infrared bands at ~1082 cm-1, and ~ 1155 cm-1, and the changes in the intensity ratios between the infrared bands at ~1025 cm-1 and ~1082 cm-1, respectively, between healthy, exfoliated cervical cell specimens and those diagnosed in a conventional, cytological manner as malignant,
Figure 6 shows the infrared spectra depicted in Figure 2, for the bands at "-1240 cm , enlarged and superimposed,
Figure 7 shows the corresponding third power derivative spectra of the infrared spectra depicted in Figure 2 for the bands in the frequency region 1190 to 1275 cm"1.
Figure 8 shows a comparison of the pressure dependencies of the vibrational frequencies of PO2 groups of nucleic acids in exfoliated cells of cervical tissues,
Figure 9 is an enlargement of the infrared bands in the frequency region 1140 cm-1 to 1185 cm-1 between normal, healthy exfoliated cervical cell specimens and those diagnosed in a conventional, cytological manner as malignant,
Figure 10 shows the corresponding third power derivative spectra of the infrared spectra shown in Figure 9, Figure 11 shows a comparison of the pressure dependancies of the frequencies of the component bands of the C-0 stretching modes in the frequency range 1140 cm-1 to 1190 cm"1 between normal, healthy, exfoliated cervical tissue specimens and those diagnosed in a conventional, cytological manner as malignant,
Figure 12 shows a comparison of the pressure dependancies of the CH? bending modes of the methylene chains in membrane lipids between normal exfoliated cervical tissue specimens and those diagnosed in a conventional, cytological manner as malignant,
Figure 13 shows a comparison of the infrared spectra in the frequency range 950 cm-1 to 1350 cm-1 for exfoliated cervical specimens diagnosed in a conventional, cytological manner as normal, malignant and dysplasia, and
Figure 14 shows infrared spectra in the frequency range 950 to 1,350 cm , obtained from healthy, exfoliated vaginal cell specimens and those diagnosed in a conventional, cytological manner as malignant.
Modes For Carrying Out The Invention
Referring now to Figure 1, there is shown an infrared source 1, a lens 2, a sample cell and holder 4, an infrared spectrometer 6, a computer 8 and a readout 10. In operation, an exfoliated tissue or cell specimen is placed in the sample cell and holder 4 and a beam of infrared light from the source 1 which has been condensed by the lens 2, is passed through the sample in the sample cell and holder 4. Any infrared absorption by the specimen is detected by the infrared spectrometer 6, which in turn is computed by the computer to give a readout at the readout 10. The computer readout may be arranged to directly indicate whether the specimen is a normal, healthy one or one which contains an anomaly (i.e. benign, dysplasia or malignant).
In tests to verify the present invention, infrared (IR) spectra were obtained as described with reference to Fig. 1 from exfoliated cervical cell specimens, that were collected from 25 females undergoing a surveillance Papanicolaou test (cervical cytology) following a standard "scraping" or "brushing" procedure. These specimens consisted of epithelial cells, inflammatory cells, red blood cells and bacteria that are part of the normal flora. The tests indicated that non-epithelial cells, present in varying numbers in all specimens and semiquantitated by two independent observers, did not influence the infrared spectra to any significant extent in the spectral regions under study; hemolysis of the red cells studied in control specimens did not significantly alter the infrared spectra either in the spectral region under study.
In further tests, cells from exfoliated scrapings obtained with a brush, were harvested by gently shaking the brush in a normal saline solution. Two halves of this cell suspension were separately pelleted by centrifugation. Following removal of the supernatant, one pellet was frozen in liquid nitrogen, to be studied later spectroscopically for IR absorption, according to the present invention, while aliquots of the other pellet were spread on microscope slides, fixed and stained with Papanicolaou stain and diagnosed in the conventional manner. Each sample was rated independently by two experienced physicians, and there was complete agreement between these ratings. Infrared absorption spectra were obtained as previously described with reference to Figure 1. Cells were smeared on the sample holder; the spectroscopic patterns were stable for many hours following sample application. An auto scale plotting configuration device was set to automatically normalize peak intensities.
The tests made fell into two groups, which were i) atmospheric pressure tests, where the sample holder described and claimed in United States Patent No. 4,980,551, dated December 25, 1990, was found to be useful, and ii) high pressure tests, where the sample holder described and claimed in United States Patent No. 4,970,396, dated November 13, 1990, was found to be useful.
Cervical specimens rated as normal healthy specimens on the basis of conventional, cytological evaluation displayed essentially identical IR spectra. The spectra of specimens diagnosed cytologically as malignant were markedly different from those of normal specimens. In Figure 2 the results for normal healthy cervical tissue are shown as , and the results for malignant cervical tissue are shown as , these are typical spectra from normal healthy and malignant cervical specimens. The most prominent differences include the following findings in malignant specimens: a) severe change in the intensity of the bands at 1025 cm-1, 1047 cm-1 (it appears as a shoulder on'- 1025 cm"1 band), 1082 cm"1, - 1155 cm-1, -1244 cm"1 and -1303 cm"1, b) significant shift of ~ 1082 cm"1, » 1155 cm"1 and —1244 cm"1 peaks, and c) a new band peaking at «_ 970 cm-1.
In Figures 3 to 5, 0 are the results for normal healthy exfoliated cervical tissue and • are the results for malignant, exfoliated cells of cervical tissue. The shifts in the two peaks at — 1082 cm"1 and -*- 1155 cm"1 averaging about 4 and 12 wave numbers, respectively, for all the specimens studied are shown in Figures 3 and 4. Figure 5 shows that the ratio of the peak intensity between the ~ 1025 cm-1 and ~ 1082 cm"1 bands differs greatly between normal and cancer tissue specimens.
The bands at ~ 1025 cm-1 and ~ 1047 cm"1 were found to be due to carbohydrates, and glycogen was found to make an overwhelming contribution to the intensity of these bands. The bands at «- 1082 cm"1 and rm 1244 cm"1 were found to be due to the symmetric and asymmetric phosphate (POj) stretching modes, respectively. They were found to originate mainly in the vibrations of the phosphodiester groups of cellular nucleic acids. The band at r*> H55 cm" was found to be due to the C-0 stretching mode of the C-OH groups of, among others, the residues of the amino acids serine, threonine and tyrosine in cell proteins as well as the C-OH groups of carbohydrates. The band at •-*' 1303 cm-1 consists of many overlapping bands and its assignment has yet to be confirmed. Finally, the band at <**• 970 cm was found to be due partly to the vibrational mode of dianionic phosphate groups, as had been previously observed in the infrared spectra of model phosphorylated proteins, and partly due to DNA and/or RNA molecules.
Referring now to Figure 6, which is an enlargement of a portion of Figure 2, and the corresponding third power derivative spectra shown in Figure 7, a more detailed study of the asymmetric phosphate stretching band revealed that in malignant, exfoliated cervical tissues, shown , it consisted of two overlapping bands, one of which, the lower frequency band, was markedly increased when compared to normal healthy, exfoliated cells. The frequency of the asymmetric PO stretching band was known at the time that the tests were made to be -*- 1220 cm"1, when the Pθ2~ group was fully hydrogen-bonded, and greater than 1240 cm"1 when it is not hydrogen-bonded. This finding indicated that in cervical cancer many Pθ2~ groups of nucleic acids are hydrogen-bonded, in contrast to those of the normal, exfoliated cervical tissue. This conclusion is supported further by the pressure dependence of the frequencies of these bands, as shown in Figure 8, where 0 shows the results for normal healthy, exfoliated cervical tissue and • the results for cancerous, exfoliated cervical tissue. The frequency of the low-frequency band was found to decrease with increasing pressure, whereas that of the high-frequency band of cancer and the single Pθ2~ band of normal healthy tissue were found to increase with increasing pressure. This response of the low-frequency band to pressure is known to be typical of a hydrogen-bonded group because pressure enhances the strength of hydrogen bonds and thereby decreases the frequency of the Pθ2~ stretching. On the other hand, the effect of pressure on the high-frequency Pθ2~ band, common to malignant and normal tissue, is known to be characteristic of non-hydrogen bonded functional groups and results from pressure enhancement of intermolecular interactions.
A similar analysis of the C-O stretching band at 1155 cm"1, shown in Figures 9 and 10 where the results for normal, healthy, exfoliated cells of cervical tissue are shown and the results for cancerous, exfoliated cells of cervical tissue are shown , revealed that it consists of three overlapping bands, 1153 cm-1, ~ 1161 cm"1 and r- H72 cm"1 of which the first two display decreased intensity and the third increased intensity when compared to the corresponding bands of normal cervical tissue. The frequencies of the first two decrease with increasing pressure, whereas that of the latter was found to increase as shown in Figure 11. These findings, therefore, indicate that the component bands at— 1153 cm"1 and «* 1161 cm" arise from the stretching vibrations of hydrogen-bonded C-OH groups, whereas the band at ~ 1172 cm-1 is due to the stretching vibrations of non hydrogen-bonded C-OH groups.
In Figure 12, where 0 shows the results for normal, healthy, exfoliated cells of cervical tissue and • shows the results for cancerous, exfoliated cells of cervical tissue, the pressure dependence of the frequency of the CH2 bending mode of the methylene chains of lipids is shown in the frequency range -— 1466 cm"1 to «*-_ 1476 cm"1. The pressure dependence in the frequency of this band has previously been widely used to study interchain packing and order/disorder properties of lipid bilayers. In lipid bilayers pressure was found to produce an increase in the frequency of this band because induced conformational and orientational ordering of the methylene chains occurred, which thereby increase interchain interactions. In malignant cervical tissue, pressure was found to induce a smaller shift in this frequency range as compared to normal healthy tissue. This indicates that in exfoliated cells of cervical cancer tissue the methylene chains of lipids are more disordered than in normal, healthy, exfoliated cells of cervical tissue.
In Figure 13, the results for normal, healthy, exfoliated cervical specimens are shown , for dysplasia are shown - • - ■ , and for malignant specimens are shown .
In Figure 13, the prominent differences for dysplasia specimens as compared to normal specimens, include: a) severe change in the intensity of the bands at 1025 cm"1, ~* 1082 cm-1, <-- 1244 cm"1 and 1303 cm"1, and b) a significant shift in the frequency of the bands at <•<- 1047 cm-1 and ~ 1082 cm-1. Also in Figure 13, the prominent differences for malignant specimens as compared to normal and dysplasia specimens include: a) severe change in the intensity of the bands at 970 cm"1, -^ 1025 cm"1, ~ 1047 cm"1, ~ 1082 cm"1, -- 1155 cm"1, 1244 cm"1 and r~* 1303 cm"1, and b) a significant shift in the frequency of the bands at -*• 1025 cm"1, -_- 1047 cm-1,
~ 1082 cm-1 and -*-• 1155 cm"1. These tests indicated that several important changes are associated with cervical, exfoliated cancer tissue. The amount of glycogen was found dramatically decreased in exfoliated, cervical cells of cancer tissue as compared to normal, healthy, exfoliated cells of cervical tissue. This is further confirmed by the fact that cancer of the liver, an organ extremely rich in glycogen, displays a similar spectroscopic pattern which is not observed in cancer of the colon, an intestinal portion poor in glycogen. The known spectrum of glycogen (D-glycogen from mammalian liver) is comparable with that obtained by the applicants in the frequence region <-> 975-1100 cm (Figures 13 and 14). Furthermore, the hydrogen-bonding of the phosphodiester groups of nucleic acids was found to increase in exfoliated cells of cervical cancer tissue.
This spectroscopic finding was common to those found in other tests with cancerous tissue biopsied specimens that the applicants had examined such as colon, liver, skin, vagina and breast.
In Figure 14, the results for healthy vaginal specimens are shown , while those for malignant specimens are shown .
In Figure 14, the prominent differences for the malignant specimens as compared to normal specimens include: a) a severe change in the intensity of the bands at <~ 1025 cm"1, _» 1154 cm"1,
Λ- 1240 cm"1 and 1300 cm"1, and b) a significant shift in the frequency of the bands at 1025cm , 1155 cm and
< 1081 cm"1. These and other tests have indicated that the present invention is useful for, for example, detecting anomalies in exfoliated tissue or cell specimens from secretions, exudates, transudates of various human or other mammalian organs or tissues, and is particularly useful for detecting exfoliated cancerous tissues or cells from Papanicolaou cervical smears, i.e., endocervical smears, ectocervical smears, vaginal smears and uterus smears.
Exfoliated cell anomalies which may be detected according to the present invention include, for example, viruses, bacteria, fungi and other infections and non-infectious diseases.
The cell anomalies may be detected in exfoliated cells or in cells in exfoliated tissue.

Claims

WE CLAIM
1. A method of detecting the presence of anomalies in exfoliated cells using infrared spectroscopy, comprising: a) directing a beam of infrared light at a specimen of exfoliated cells, and b) determining if an anomaly is present in the specimen by spectral analysis of the infrared absorption by the specimen, at at least one range of frequencies, to ascertain whether at least one change in the infrared absorption characteristics has occurred, due to vibration of at least one functional group of molecules, present in the specimen, which is characteristic of that anomaly.
2. A method according to claim l, wherein the said at least one change in the infrared absorption characteristics is a change in absorption intensity at a particular frequency, a change of frequency at which a particular absorption occurs, or a different pressure applied to the functional group causing a change of frequency at which a particular absorption occurs.
3. A method according to claim 1, wherein the functional group of molecules is in at least one of the following molecules, carbohydrates, nucleic acids, tissue proteins or membrane lipids.
4. A method according to claim 1, wherein the functional group is a phosphodiester group in nucleic acids, a C-OH group in tissue protein and carbohydrates, or a CH2 group in lipids.
5. A method according to claim 1, wherein the functional group of molecules is in at least one of the ollowing: carbohydrates, phosphodiester groups in nucleic acids,
C-OH groups of tissue proteins and carbohydrates, or
CH„ group of lipids.
6. A method according to claim 1, wherein the specimen is a Papanicolau smear.
7. A method according to claim 1, wherein the specimen is a cervical specimen.
8. A method according to claim 1, wherein the specimen is an endocervical specimen.
9. A method according to claim 1, wherein the specimen is an ectocervical specimen.
10. A method according to claim 1, wherein the specimen is a vaginal specimen.
11. A method according to claim 1, wherein the specimen is a uterus specimen.
EP92903692A 1991-02-04 1992-01-31 A method of detecting the presence of anomalies in exfoliated cells using infrared spectroscopy Withdrawn EP0570418A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CA 2035603 CA2035603C (en) 1991-02-04 1991-02-04 A method of detecting the presence of anomalies exfoliated cells using infrared spectroscopy
CA2035603 1991-02-04

Publications (1)

Publication Number Publication Date
EP0570418A1 true EP0570418A1 (en) 1993-11-24

Family

ID=4146947

Family Applications (1)

Application Number Title Priority Date Filing Date
EP92903692A Withdrawn EP0570418A1 (en) 1991-02-04 1992-01-31 A method of detecting the presence of anomalies in exfoliated cells using infrared spectroscopy

Country Status (6)

Country Link
EP (1) EP0570418A1 (en)
JP (1) JPH06507237A (en)
CN (1) CN1067314A (en)
AU (1) AU1180892A (en)
CA (1) CA2035603C (en)
WO (1) WO1992014134A1 (en)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5733739A (en) * 1995-06-07 1998-03-31 Inphocyte, Inc. System and method for diagnosis of disease by infrared analysis of human tissues and cells
WO1997014961A1 (en) * 1995-10-18 1997-04-24 Aktsionernoe Obschestvo Zakrytogo Tipa 'ntf Intersvyaz' Method of diagnosing malignant disorders
AUPN825796A0 (en) 1996-02-26 1996-03-14 Ashdown, Martin The application of infrared (ir) spectrometry to the investigations of components of blood and other body fluids
JP2002508845A (en) 1997-06-27 2002-03-19 パシフィック ノースウェスト リサーチ インスティテュート How to distinguish between metastatic and non-metastatic tumors
JP4403908B2 (en) * 2004-07-27 2010-01-27 住友電気工業株式会社 Peptide or protein phosphorylation analyzer, phosphorylation discrimination program, and recording medium for the program
WO2011151825A2 (en) 2010-06-01 2011-12-08 Todos Medical Ltd. Diagnosis of cancer
EP2707710B1 (en) 2011-05-11 2022-08-17 Todos Medical Ltd. Diagnosis of cancer based on infrared spectroscopic analysis of dried blood plasma samples
US9804145B2 (en) 2013-05-28 2017-10-31 Todos Medical Ltd. Infrared analysis of benign tumors
KR101735464B1 (en) 2014-12-17 2017-05-15 한밭대학교 산학협력단 Device for detecting organic material and measuring death rate of cells, and method thereof
GB2545877B (en) * 2015-09-10 2021-09-15 Sierra Medical Ltd ATR-FTIR computational analysis of Barrett's esophagus and esophageal cancers
JP2017203637A (en) * 2016-05-09 2017-11-16 住友電気工業株式会社 Tumor cell detection method and tumor cell detection device
CN110702720A (en) * 2019-10-09 2020-01-17 李俊菲 Sampling swab and collection set for gynecological secretion analyzer

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4017192A (en) * 1975-02-06 1977-04-12 Neotec Corporation Optical analysis of biomedical specimens
DE3903777A1 (en) * 1989-02-09 1990-08-16 Bruker Analytische Messtechnik METHOD FOR THE QUICK DETECTION OF MICROORGANISMS IN SAMPLES AND DEVICE FOR IMPLEMENTING THE METHOD
US4970396A (en) * 1989-12-26 1990-11-13 National Research Council Of Canada/Conseil National De Recherches Du Canada Infrared spectra recording, high pressure sample holder
US4980551A (en) * 1990-01-05 1990-12-25 National Research Council Canada Conseil National De Recherches Canada Non-pressure-dependancy infrared absorption spectra recording, sample cell
CA2008831C (en) * 1990-01-29 1996-03-26 Patrick T.T. Wong Method of detecting the presence of anomalies in biological tissues and cells in natural and cultured form by infrared spectroscopy

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9214134A1 *

Also Published As

Publication number Publication date
CN1067314A (en) 1992-12-23
CA2035603A1 (en) 1992-08-05
CA2035603C (en) 1998-08-04
WO1992014134A1 (en) 1992-08-20
AU1180892A (en) 1992-09-07
JPH06507237A (en) 1994-08-11

Similar Documents

Publication Publication Date Title
US5168162A (en) Method of detecting the presence of anomalies in exfoliated cells using infrared spectroscopy
Ci et al. Fourier transform infrared spectroscopic characterization of human breast tissue: implications for breast cancer diagnosis
Chiriboga et al. Infrared spectroscopy of human tissue. I. Differentiation and maturation of epithelial cells in the human cervix
CA2008831C (en) Method of detecting the presence of anomalies in biological tissues and cells in natural and cultured form by infrared spectroscopy
Wong et al. Pressure-tuning FT-IR study of human cervical tissues
CA2035603C (en) A method of detecting the presence of anomalies exfoliated cells using infrared spectroscopy
Krafft et al. Analysis of human brain tissue, brain tumors and tumor cells by infrared spectroscopic mapping
Maziak et al. Fourier-transform infrared spectroscopic study of characteristic molecular structure in cancer cells of esophagus: an exploratory study
Wong et al. Characterization of exfoliated cells and tissues from human endocervix and ectocervix by FTIR and ATR/FTIR spectroscopy
Wong et al. Infrared spectra of microtome sections of human colon tissues
AU2011333336B2 (en) Methods and kits for differential staining of abnormal urinary system cells
US20050250091A1 (en) Raman molecular imaging for detection of bladder cancer
Bandzeviciute et al. Fiber based infrared spectroscopy of cancer tissues
RU2740431C1 (en) Method of preparing cell blocks based on cervical exfoliative material and cervical canal
JPH10505412A (en) Infrared spectroscopy of preservative treated samples
Wang et al. Evaluation of gallbladder lipid level during carcinogenesis by an infrared spectroscopic method
SU1114004A1 (en) Method of diagnostics of degree of malignization in case of fibrous-cystoid mastopathy
RU2670655C2 (en) Method for determining the risk of recurrence of superficial bladder cancer after surgical treatment
RU2104526C1 (en) Method for diagnosing t-cellular lymphomae of skin
RU2235323C2 (en) Method of diagnosing malignant tumors
RU197467U1 (en) BIOCHIP FOR MULTIPLEX ANALYSIS
Törnberg et al. Fluorescence microscopy and acridin-orange staining in the cytological diagnosis of atypical changes in cervical epithelium
RU2092843C1 (en) Method of differential diagnosis of fibroadenoma and mamma cancer
CA2126915A1 (en) Method of detecting the presence of anomalies in a preservative treated sample, prepared from fresh biological tissue or cells in natural or cultured form, by infrared spectroscopy
Shirazi et al. Fourier-Transform Infrared Spectroscopic Comparison of Normal and Malignant Cervical Tissue Cervical Tissue.

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19930827

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): DE FR GB IT

17Q First examination report despatched

Effective date: 19931217

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 19951116