Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2000-07-24
2004-06-08
Forman, B J (Department: 1634)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S174000, C435S283100, C435S287200, C422S050000, C422S068100
Reexamination Certificate
active
06746840
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to a micro array and an analyzing method using the micro array. This invention particularly relates to a micro array, which comprises a base plate and multiple kinds of biomolecules arrayed and fixed on the base plate. This invention also relates to an analyzing method using the micro array, wherein the micro array is subjected to hybridization with a solution containing a labeled biomolecule, and a biomolecule fixed on the base plate of the micro array, which biomolecule has been hybridized with the labeled biomolecule, is specified.
As will be described later, the term “micro array” as used herein has broad meanings embracing a micro array, a macro array, a DNA chip, and others.
2. Description of the Prior Art
A thesis entitled “DNA microarray for gene expression analysis” is published in Experimental Medicine Series, Yodosha Co., Vol. 17, the January 1999 issue, pp. 61-65. In the thesis, a technique for performing a genetic expression analysis by the utilization of a micro array is explained in detail.
Recently, the genetic expression analyzing techniques utilizing micro arrays have widely been used in practice. As illustrated in
FIG. 6
, in the genetic expression analyzing techniques, a micro array comprising a base plate
40
and multiple kinds of biomolecules arrayed and fixed in a matrix-like form on the surface of the base plate
40
is utilized. The base plate
40
is constituted of a membrane, a glass, a slide glass, a silicon base plate, or the like. As the multiple kinds of biomolecules, currently, cDNA, oligo-DNA, other DNA's, PNA, EST, and the like, are utilized. The micro array comprising the base plate
40
and multiple kinds of biomolecules arrayed and fixed in a matrix-like form on the surface of the base plate
40
is referred to as the macro array, the micro array, the DNA chip, or the like, depending on the kind of the base plate
40
, and the like. In this specification, the group of the macro array, the micro array, the DNA chip, and the like, is generically referred to as the “micro array.”
Also, in the genetic expression analyzing techniques utilizing micro arrays, various kinds of biomolecules, such as cDNA, genome DNA, mRNA, total RNA, other RNA's, dNTP, and PNA, which have been labeled with a radioactive isotope, a fluorescent substance, or the like, are prepared.
Thereafter, the biomolecules, which have been fixed in a matrix-like form on the surface of the base plate
40
, and a biomolecule, which has been labeled with the radioactive isotope, the fluorescent substance, or the like, are subjected to hybridization.
In cases where the biomolecules having been fixed on the surface of the base plate
40
contains a biomolecule, which is capable of undergoing hybridization (binding) with the biomolecule having been labeled with the radioactive isotope, the fluorescent substance, or the like, the fixed biomolecule and the labeled biomolecule are hybridized with each other on the base plate
40
. As a result, the radioactive isotope, the fluorescent substance, or the like, is fixed at a position on the base plate
40
, at which the fixed biomolecule having been hybridized with the labeled biomolecule is located. Also, the radioactive isotope, the fluorescent substance, or the like, is not fixed at positions on the base plate
40
, at which the fixed biomolecules having not been hybridized with the labeled biomolecule are located. In
FIG. 6
, the circles surrounding the dots indicate the positions on the base plate
40
, at which the fixed biomolecule having been hybridized with the labeled biomolecule is located, i.e. the positions at which the radioactive isotope, the fluorescent substance, or the like, has been fixed to the base plate
40
.
FIG. 6
is a conceptual view, in which the dots located in a matrix-like form can be visually discriminated from one another. However, actually, fine dots are located at a high density on the base plate
40
, and therefore they cannot be visually discriminated from one another.
In cases where the label is the radioactive isotope, a stimulable phosphor sheet capable of storing energy from radiation thereon may be utilized in order to detect the positions on the base plate
40
at which the radioactive isotope is located.
When the stimulable phosphor sheet is exposed to radiation radiated out from the radioactive isotope, the stimulable phosphor sheet stores energy from the radiation. When the stimulable phosphor sheet, on which the energy from the radiation has been stored, is exposed to stimulating rays, such as a laser beam, which cause the stimulable phosphor sheet to emit light in proportion to the amount of energy stored thereon during its exposure to the radiation, light is emitted by the stimulable phosphor sheet. At this time, no light is emitted from the sites, which were not exposed to the energy from the radiation. One of typical stimulable phosphor sheets comprises a substrate and a stimulable phosphor layer, which is overlaid on the substrate and comprises a binder and BaFX phosphor particles dispersed at a high density in the binder, where X represents a halogen. The stimulable phosphor sheet is also known as a radiation image storage panel utilizing a stimulable phosphor.
As illustrated in
FIG. 7
, the stimulable phosphor sheet (also referred to as the imaging plate) IP is brought into close contact with the surface of the base plate
40
, on which the radioactive isotope has been locally fixed as a result of the hybridization. In this manner, the stimulable phosphor sheet is exposed locally to the radiation radiated out from the radioactive isotope. In
FIG. 7
, the base plate
40
is turned upside down from the state of FIG.
6
and is then brought into close contact with the stimulable phosphor sheet IP.
As a result, the stimulable phosphor sheet IP is exposed locally to the radiation. The stimulable phosphor sheet IP, on which the energy from the radiation has been stored, is then exposed to the stimulating rays and is caused to locally emit light. In accordance with the position which emits the light, the position on the base plate
40
, at which the fixed biomolecule having been hybridized with the labeled biomolecule is located, can be specified. Also, in accordance with the thus specified position, the kind of the fixed biomolecule, which has been hybridized with the labeled biomolecule, can be specified.
FIG. 8
is an explanatory view showing the read-out step, which is performed in the manner described above. In
FIG. 8
, the circle indicates the site on the stimulable phosphor sheet IP, which was locally exposed to the radiation coming from the radioactive isotope and has stored the energy from the radiation. Also, in
FIG. 8
, reference numeral
41
represents the stimulating rays, and reference numeral
42
represents the light emitted from the site on the stimulable phosphor sheet IP.
When the stimulable phosphor sheet IP is exposed to the stimulating rays and is caused to emit the light, the site at which the energy from the radiation has been stored returns to the state in which no energy from the radiation was stored. Therefore, the stimulable phosphor sheet IP can be used repeatedly. However, as illustrated in
FIG. 7
, with the conventional technique described above, the processing must be performed for accurately setting the position of the stimulable phosphor sheet IP with respect to the base plate
40
, and bringing the entire area of the stimulable phosphor sheet IP into close contact with the base plate
40
and thus superposing the entire area of the stimulable phosphor sheet IP upon the base plate
40
. Therefore, the conventional technique described above has the problems in that considerable time and labor are required to perform the processing. Also, with the conventional technique described above, it is necessary to perform the step for bringing the stimulable phosphor sheet IP into close contact with the base plate
40
and causing the energy from the radiation to be stored on the
Forman B J
Fuji Photo Film Co. , Ltd.
Sughrue & Mion, PLLC
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