Patent US3001900A successfully refreshed from Google Patents.
Patent EP0214713B1 successfully refreshed from Google Patents.
EP0214713B1
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EP
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Method of analysing the results of electrophoresis, method of separating and detecting the components of a mixture, and separation apparatus
Assignee: Astromed Ltd
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Filing: 1986-05-27
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Granted: 1991-02-27
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Expires: 2006-05-27
Abstract
Abstract Light from the pattern of spots of bands resulting from electrophoresis is detected by a two-dimensional charge coupled device (26). Visualisation of the spots or bands may be achieved by viewing in transmitted light if the material subject to electrophoresis is dyed, by fluorescent markers in the material stimulated by untraviolet, or by a scintillating material if radioactive markers are used.Preferably the charge coupled device (26) is cooled to suppress thermal dark current, improving sensitivity and dynamic range.Light can be detected from moving components during separation by electrophoresis by clocking the charge coupled device to move charge packets in synchronism with movement of the components. This means that each charge packet moves through all the elements of a charge coupled device column during the detection, resulting in automatic averaging for the detection sensitivity variations in the charge coupled device column. Further, it is thereby possible to detect each separated band or spot at the same position in the gel at different times, and thus at the same degree of resolution, instead of detecting the bands or spots at different positions at the same time.
Inventors
- 👤 Limited Astromed
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-26 14:34
CPC Classifications
G; G01; G01N; G01N27/00; G01N27/26; G01N27/416; G01N27/447; G01N27/44704; G01N27/44717; G01N27/44721
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
98 words
A method of analysing samples by use of electrophoresis comprising the steps of: - treating the samples with fluorescent marking material so that components of the samples are fluorescently marked; - applying the samples to an electrophoretic gel; - running the gel to effect; electrophoresis causing differential migration of different components; - irradiating the gel with a U/V source to render the marked components visible; - detecting the pattern of light emanating from the marked components by means of a light sensitive charge coupled device (CCD), said CCD having a two-dimensional detector array and being cooled during detection.
Claim 2
Dependent Claim
22 words
A method according to claim 1, in which the samples are treated with fluorescent marking material prior to application to the gel.
Claim 3
Dependent Claim
60 words
A method according to claim 1 or 2, in which the charge coupled device is a frame transfer CCD and the pattern of light is detected during electrophoretic movement, charge packets being shifted through the CCD in the same direction as and in synchronism with movement of the pattern of light falling on the CCD due to the electrophoretic movement.
Claim 4
Dependent Claim
34 words
A method according to claim 3, in which the CCD is used to detect bands or spots contained in the pattern of light when they are brought to a predetermined location by the electrophoresis.
Claim 5
Dependent Claim
39 words
A method according to claim 1 or 2, in which the running of the gel to effect separation by electrophoresis is halted, the pattern of light from separated components is detected, and running of the gel is then resumed.
Claim 6
Independent Claim
25 words
A method according to any one of the preceding claims, in which the charge coupled device is cooled during operation to substantially below ambient temperature.
Claim 7
Dependent Claim
19 words
A method according to claim 6, in which the charge coupled device is cooled to down to about 125K.
Claim 8
Independent Claim
79 words
A method according to any one of the preceding claims, in which the samples are marked with a fluorescent material which emits electromagnetic radiation at a first wavelength and a calibration chemical is mixed with the material, the calibration chemical being marked with a fluorescent material which emits electromagnetic radiation at a second wavelength, and the wavelengths detected by the charge coupled device are varied to enable the calibration chemical to be distinguished from the material subject to electrophoresis.
Claim 9
Independent Claim
32 words
A method according to any one of the preceding claims, in which the output from the charge coupled device is compared with the results of previous electrophoretic separations stored in a databank.
Claim 10
Independent Claim
37 words
A method according to any one of the preceding claims, in which the charge coupled device is coated with one or more materials which absorb electromagnetic radiation at one wavelength and emit electromagnetic radiation at another wavelength.
Claim 11
Independent Claim
37 words
A method according to any one of the preceding claims, in which the electrophoresis is performed on four parallel one-dimensional tracks, a respective DNA segment (A, C, G or T), being subject to electrophoresis in each track.
Claim 12
Dependent Claim
99 words
Apparatus for analysing fluorescently marked samples by electrophoresis, comprising: - a source of U/V radiation for illuminating an electrophoretic gel to which the fluorescently marked samples have been applied; - means to control running of gel to effect electrophoresis, causing differential migration of different components of the samples giving rise to a pattern of light due to illumination of the gel by the U/V source; and - means for detecting the pattern of light; wherein the detecting means comprises a light sensitive charge coupled device (CCD) having a two-dimensional detector array; and means for cooling the CCD during detection.
Claim 13
Dependent Claim
12 words
Apparatus according to claim 12, in which two-dimensional electrophoresis can be performed.
Claim 14
Dependent Claim
18 words
Apparatus according to claim 12 or 13, in which the charge coupled device is a frame transfer CCD.
Claim 15
Dependent Claim
33 words
Apparatus according to claim 14, having driving circuits for the charge coupled device adapted to drive the charge coupled device in synchronism with movement of the light falling thereon due to electrophoretic movement.
Claim 16
Dependent Claim
24 words
Apparatus according to any one of claims 12 to 14, in which the cooling means can reduce the temperature to down to about 125K.
Claim 17
Dependent Claim
27 words
Apparatus according to any one of claims 12 to 16, in which means is provided to vary the wavelength of light detected by the charge coupled device.
Claim 18
Dependent Claim
37 words
Apparatus according to any one of claims 12 to 17, including a databank of results of previous electrophoresis runs and electronic means for comparing the output of the charge coupled device with the content of the databank.
Claim 19
Dependent Claim
37 words
Apparatus according to any one of claims 12 to 18, in which the charge coupled device is coated with one or more materials which absorb electromagnetic radiation of one wavelength and emit electromagnetic radiation of another wavelength.
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