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WO2002006880A2 - Appareil a lames a substrat plastique pour analyse microscopique aux ir - Google Patents

Appareil a lames a substrat plastique pour analyse microscopique aux ir Download PDF

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Publication number
WO2002006880A2
WO2002006880A2 PCT/CA2001/001031 CA0101031W WO0206880A2 WO 2002006880 A2 WO2002006880 A2 WO 2002006880A2 CA 0101031 W CA0101031 W CA 0101031W WO 0206880 A2 WO0206880 A2 WO 0206880A2
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WO
WIPO (PCT)
Prior art keywords
slide
infrared
plastic
range
substrate
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.)
Ceased
Application number
PCT/CA2001/001031
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English (en)
Other versions
WO2002006880A3 (fr
Inventor
Paul Anthony Presta
Michael Maszkiewicz
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.)
Veracel Inc
Original Assignee
Veracel Inc
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 Veracel Inc filed Critical Veracel Inc
Priority to AU2001275617A priority Critical patent/AU2001275617A1/en
Publication of WO2002006880A2 publication Critical patent/WO2002006880A2/fr
Publication of WO2002006880A3 publication Critical patent/WO2002006880A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides

Definitions

  • the present invention relates to medical and biological specimen analysis, and more particularly to a plastic substrate slide apparatus and technique for infrared microscopy analysis.
  • FTIR Fourier Transform Infrared
  • the mid-Infrared (i.e. 2.5 - 25 microns) response of normal and pre-cancerous human tissue comprises a variety of structural and chemical changes which may be used to discriminate between the two. These changes include increases in glycogen content, extensive hydrogen bonding of phosphodiester groups in nucleic acids, tighter physical packing of nucleic acids, phosphorylation of C-OH groups in carbohydrates and proteins, increased disorder of methylene chains in membrane lipids, increased ratio of methyl to methylene, reduction in the hydrogen bond strength in the amide groups of ⁇ -helical segments and an increase in the hydrogen bond strength in the amide groups of the ⁇ -sheet segments.
  • Proposals have been made to utilize these mechanisms in cancer screening protocols, such as, the early detection of pre-cancerous lesions of the uterine cervix and carcinomatous breast tissue.
  • Pap test a screening test that searches for evidence of precancerous lesions in exfoliated cervical cells.
  • the Pap test Involving the manual examination of tens of thousands of cervical epithelial cells, the Pap test is costly to apply and is subject to human error. Nevertheless, it has an enviable record of reducing cervical cancer mortality in the countries where it is applied. As a result considerable effort has been put into developing automated alternatives for the manual Pap test.
  • the third step is known as feature extraction.
  • feature extraction each of the segmented regions or objects in the image is subjected to a range of mathematical measures that seek to encapsulate the visual appearance in numerical form.
  • classification comprises utilizing the numerical features to arrive at some type of conclusion about the object's identity.
  • the mid- infrared absorption properties of the cervical epithelial cells may be used to yield a new and useful channel of information in addition to the usual visible-light imaging measures.
  • these new mid-IR channels may be used to improve segmentation performance, to create a new set of features for classification, or as discrimination measures on their own. It has been found that restricting the use of the mid-IR channels to regions of the samples that contain nuclear material improves the effective signal-to-noise ratio of the measurement over bulk tissue measurements offering a greater power of discrimination to the technique.
  • differential mid-IR measure one channel acting as an interrogator, the other as a reference
  • a differential mid-IR measure allows a high-speed real-time scan of cellular material to quickly discriminate between normal and abnormal cells or tissue and may be extended to other important diagnostic properties of the cells.
  • the nature of the substrate or carrier holding the specimen poses an acute problem for practical applications of mid-infrared spectroscopy to anatomic pathology.
  • the most common substrate for anatomic pathology evaluations is the glass microscope slide. Glass slides are typically manufactured from borosilicate glass to relatively relaxed tolerances.
  • the problem with borosilicate glasses (and most other reasonable substitute glasses) is the lack of a suitable transmission window for mid-infrared radiation. As shown in Fig. 4, the optical transmission of borosilicate glass declines rapidly after 1.5 microns. This is well short of the mid-IR region of interest which falls between 7 and 12 microns. Thus, the interrogation signal required by an automated system is essentially absorbed indiscriminately by the glass substrate.
  • the present invention provides an apparatus and technique for a microscope slide or carrier suitable for application of mid-IR spectroscopic techniques, such as, discrimination between normal and pre-cancerous cells prepared in the form cells or cell groupings in a specimen placed on the slide.
  • the present invention provides a microscope slide comprising a plastic substrate having mid-infrared transmissive characteristics in the range of 7 to 12 microns.
  • the present invention provides a slide apparatus for infrared microscopy analysis of a biological specimen
  • the slide apparatus comprises: (a) a plastic slide substrate for receiving the biological specimen; (b) the plastic slide substrate has optical transmission characteristics in the mid- infrared range.
  • the present invention provides a slide apparatus for infrared microscopy analysis of a cytological sample prepared according to the Pap staining protocol, the slide apparatus comprises: (a) a plastic slide substrate for receiving the cytological sample; (b) the plastic slide substrate comprises polyethylene having optical transmittance characteristics substantially in the range 975 to 1000 cm -1 .
  • the present invention provides a slide apparatus for infrared microscopy analysis of a cytological sample prepared according to the Pap staining protocol, the slide apparatus comprises: (a) a plastic slide substrate for receiving the cytological sample; (b) the plastic slide substrate comprises polyethylene having optical transmittance characteristics substantially in the range 1155 to 1180 cm "1 .
  • the present invention provides slide a apparatus for infrared microscopy analysis of a cytological sample prepared according to the Pap staining protocol, the slide apparatus comprises: (a) a plastic slide substrate for receiving the cytological sample; (b) the plastic slide substrate comprises polymethyl pentene having optical transmittance characteristics substantially in the range 975 to 1000 cm "1 .
  • the present invention provides slide a apparatus for infrared microscopy analysis of a cytological sample prepared according to the Pap staining protocol, the slide apparatus comprises: (a) a plastic slide substrate for receiving the cytological sample; (b) the plastic slide substrate comprises polymethyl pentene having optical transmittance characteristics substantially in the range 1155 to 1180 cm "1 .
  • Fig. 1 shows in diagrammatic form a plastic slide apparatus suitable for infrared microscopy analysis according to the present invention
  • Fig.2 shows in graphical form the optical transmittance characteristics for the plastic slide formed from polyethylene (PE) according to one embodiment of the invention
  • Fig.3 shows in graphical form the optical transmittance characteristics for the plastic slide formed from polymethyl pentene (TPX).
  • Fig.4 shows in graphical form the optical transmittance characteristics for a conventional borosilicate glass specimen slide.
  • Fig. 1 shows in diagrammatic form a plastic slide 10 suitable for infrared microscopy analysis in accordance with the present invention.
  • the slide 10 comprises a substrate or carrier indicated by reference 12. Cells indicated by reference 20 are deposited on the slide substrate 12. The cells 20 may be covered by a plastic cover slip 14.
  • the plastic cover slip 14 is formed from the same material as the substrate 12. As will be described in more detail below, the plastic slide 10 provides optical transmission in the mid-infrared region in the range of 7 to 12 microns.
  • the cells 20 may be stained for the usual visual analysis (i.e. the Papanicolaou procedure for epithelial cell observations) and mounted in the usual mounting medium in an automated analysis system for example.
  • visual analysis i.e. the Papanicolaou procedure for epithelial cell observations
  • the cells in the specimen 20 that respond to the stain comprise naturally- derived, encapsulated protein and this is what the stains will generally bind to.
  • the hematoxylin will bind to DNA or RNA in the cell nucleus
  • the Orange G stain will selectively bind to keratin in the cytoplasm, etc.
  • an infrared absorbance signal at 975 cm “ 1 to 1000 cm “1 and again at 1155 cm “1 to 1180 cm “1 may be used to discriminate between normal and abnormal cells.
  • the infrared band between 975 cm “1 to 1000 cm “1 corresponds to the symmetric stretching vibration of the dianionic phosphate monoesters of phosphorylated proteins and nucleic acids.
  • the infrared band at 1155 cm “1 to 1180 cm “1 is associated with the stretching vibrational modes of the C-O groups serine, threonine, or tyrosine amino acid residues of cellular proteins.
  • the substrate 12 for the plastic slide 10 is manufactured from polyethylene (PE).
  • PE polyethylene
  • the cellular sample 20 to be analyzed is fixed using known techniques and then deposited onto the surface of the plastic slide 10.
  • Fig. 2 shows in graphical form optimal transmittance characteristics for polyethylene.
  • polyethylene has an optical transmission in the mid-IR 'windows' or bands of 975 - 1000 cm "1 and 1155 - 1180 cm "1 which is never less than 50%.
  • the substrate 12 for the plastic slide 10 is made from polymethyl pentene (TPX). As shown in Fig. 3, the substrate 12 of polymethyl pentene shows an optical transmission of more than 20% in the mid-IR bands of 975 - 1000 cm “1 and 1155 - 1180 cm “1 .
  • TPX polymethyl pentene
  • a cell or cells 20 deposited on the plastic slide 10 are analyzed according to the following method.
  • the cell or cells 20 of interest on the slide 10 are brought into focus using conventional light microscopy by either automatic or manual means.
  • the microscope stage is translated to a blank region that contains no cells at all.
  • a background reading is obtained for the infrared absorption through the substrate 12 of the plastic slide 10 using an infrared microscopy apparatus (not shown).
  • the background infrared bands of interest lie between 970 cm “1 and 1000 cm “1 and again between 1145 cm “1 and 1190 cm “1 .
  • the slide 10 carrying the sample or specimen is rapidly scanned with the infrared microscope (not shown) across all of the cells in the specimen.
  • Scan data is collected in the infrared bands of 970 cm “1 and 1000 cm “1 and 1145 cm “1 and 1190 cm “1 .
  • the absorbence is corrected for the presence of the background, and the following ratios are calculated:
  • the plastic slide according to the invention provides apparatus which increases the signal-to-noise ratio of the relatively weak mid-IR absorbencies and makes possible the practical realization of mid-infrared microscopy analysis for cytological specimens.

Landscapes

  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

L'invention porte sur un appareil à lames à substrat plastique pour analyse microscopique aux IR. Le substrat plastique est constitué d'un matériau dont les caractéristiques de transmission optique se situent dans l'IR moyen ou dans la plage de 7 à 12 microns. Dans une version, le substrat plastique, fait de polyéthylène (PE), assure une transmission optique dans les bandes de l'IR moyen de 975 à 1000 cm-1 et de 1155 à 1180 cm-1. Dans une autre version, le substrat plastique, fait de polyméthyl pentène (TPX), assure également une transmission optique dans les bandes de l'IR moyen de 975 à 1000 cm-1 et de 1155 à 1180 cm-1.
PCT/CA2001/001031 2000-07-17 2001-07-16 Appareil a lames a substrat plastique pour analyse microscopique aux ir Ceased WO2002006880A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2001275617A AU2001275617A1 (en) 2000-07-17 2001-07-16 Plastic substrates slide apparatus for infrared microscopy analysis

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US21855700P 2000-07-17 2000-07-17
US60/218,557 2000-07-17

Publications (2)

Publication Number Publication Date
WO2002006880A2 true WO2002006880A2 (fr) 2002-01-24
WO2002006880A3 WO2002006880A3 (fr) 2002-10-24

Family

ID=22815567

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2001/001031 Ceased WO2002006880A2 (fr) 2000-07-17 2001-07-16 Appareil a lames a substrat plastique pour analyse microscopique aux ir

Country Status (2)

Country Link
AU (1) AU2001275617A1 (fr)
WO (1) WO2002006880A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103105670A (zh) * 2013-02-06 2013-05-15 李宏 镜下观察用细胞载片

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5095213A (en) * 1988-11-03 1992-03-10 Syntex (U.S.A.) Inc. Method of using an opaque plastic microscope slide for epi-fluorescent microscopy
JPH07500180A (ja) * 1991-06-25 1995-01-05 ミネソタ・マイニング・アンド・マニュファクチュアリング・カンパニー 分光器用のサンプルホルダーおよびその使用方法
US5463223A (en) * 1994-01-24 1995-10-31 Patwong Technologies, Inc. Disposable all purpose micro sample holder
JP2000501844A (ja) * 1995-07-19 2000-02-15 モルフォメトリックス テクノロジーズ インク． 顕微鏡スライドの自動走査
US5945674A (en) * 1997-07-30 1999-08-31 Vysis, Inc. Method of identifying cellular types in a biological sample supported on an absorptive substrate by infrared spectroscopy
US5812312A (en) * 1997-09-04 1998-09-22 Lorincz; Andrew Endre Microscope slide

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103105670A (zh) * 2013-02-06 2013-05-15 李宏 镜下观察用细胞载片

Also Published As

Publication number Publication date
AU2001275617A1 (en) 2002-01-30
WO2002006880A3 (fr) 2002-10-24

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