Signal processing method in autoradiography

• US 4,888,695 A
• Filed: 01/06/1984
• Issued: 12/19/1989
• Est. Priority Date: 01/08/1983
• Status: Expired due to Term

• Current Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Original Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Inventors: Kimura, Tsutomu, Hishinuma, Kazuhiro, Shiraishi, Hisashi
• Primary Examiner(s): Smith, Jerry
• Assistant Examiner(s): Meyer, Charles B.
• Application Number:
  US06/568,876
• Time in Patent Office: 2,174 Days
• Field of Search: 364/414, 364/413, 382/6, 435/6, 435/76, 435/77, 436/169, 436/174
• US Class Current: 382/129
• CPC Class Codes: G01T 1/2012 using stimulable phosphors, e.g. stimulable phosphor sheets
• CPC Class Codes: G01T 1/2014 Reading out of stimulable sheets, e.g. latent image

Abstract

A signal processing method in autoradiography for determination of the base sequence of DNA or a DNA fragment, employing groups of radioactively labeled base specific cleavage products or mixtures thereof obtained by specifically cleaving the DNA or DNA fragment labeled with a radioactive element and resolved one-dimensionally in parallel relation to each other to form resolved rows on a support medium. An autograph is obtained having locational information on groups of radioactively labeled cleavage products contained in the rows on the support medium. An electrical digital signal corresponding to the autoradiograph are generated by recording and reproducing a radiation image stored in a stimulable phosphor sheet. Sampling points in each resolved row of the digital signal are next detected. Reference sampling points are then determined in a plurality of reference rows which are either directly provided on the support medium and/or synthesized from certain resolved rows. Corresponding reference sampling points in the plural reference rows are joined to generate plural continuous lines comprising straight lines, polygonal lines or curved lines. Sampling points in the remaining non-reference rows are identified by comparison with the continuous lines to thus determine the base sequence of the DNA or DNA fragment.

Claims

1. A signal processing method in autoradiography for determination of the base sequence of DNA or a DNA fragment, employing radioactively labeled cleavage products obtained by specifically cleaving the DNA or DNA fragment labeled with a radioactive element, comprising the steps of:

(1) providing at least two sets of reference mixtures each consisting essentially of guanine specific cleavage products, adenine specific cleavage products, thymine specific cleavage products and cytosine specific cleavage products;

(2) providing at least one group of cleavage products or a mixture thereof containing at least one kind of the base specific cleavage products;

(3) resolving both the reference mixtures provided in step (1) and the product group provided in step (2) one-dimensionally in such parallel relation that the resolved rows of said two sets of reference mixtures (1) sandwich at least one resolved row of the product group (2) to form at least three resolved rows on a support medium;

(4) obtaining an autoradiograph having locational information on groups of radioactively labeled cleavage products contained in said rows on the support medium;

(5) generating an electrical digital signal corresponding to said autoradiograph including said rows

(6) detecting reference sampling points in each resolved reference row of the digital signal;

(7) detecting sampling points in said one resolved row;

(8) joining corresponding reference sampling points in said reference rows to generate plural continuous lines comprising straight lines, polygonal lines or curved lines; and

(9) identifying said sampling points in said one resolved row of the product group (2) by comparison of the last-mentioned sampling points with said continuous lines to thus determine said base sequence of said DNA or DNA fragment.

(9) causing a stimulable phosphor sheet to absorb radiation and;

(10) scanning said stimulable phosphor sheet with an electromagnetic wave to release at least a portion of radiation energy stored in said phosphor sheet in the form of the autordiograph as stimulated emission, and detecting said stimulated emission photoelectrically.

2. The signal processing method in autoradiography as claimed in claim 1, wherein at least three resolved rows of the reference mixture (1) are so provided on the support medium that two resolved rows of the reference mixtures in close relation to each other sandwich at least one resolved row of the product group (2).

3. The signal processing method in autoradiography as claimed in claim 1 or 2, wherein the sampling points are detected by at least smoothing or threshold-processing the digital signal one each scanning line for the resolved rows respectively.

4. The signal processing method in autoradiography as claimed in claim 1 or 2, wherein the employed cleavage products of DNA or the DNA fragment comprise at least five groups including:

(1) guanine specific cleavage products and adenine specific cleavage products and thymine specific cleavage products and cytosine specific cleavage products

(2) base specific cleavage products containing guanine specific cleavage products;

(3) base specific cleavage products containing adenine specific cleavage products;

(4) base specific cleavage products containing thymine specific cleavage products; and

(5) base specific cleavage products containing cytosine specific cleavage products.

5. A signal processing method in autoradiography for determination of the base sequence of DNA or a DNA fragment, employing radioactively labeled cleavage products obtained by specifically cleaving the DNA or DNA fragment labeled with a radioactive element, comprising the steps of:

(1) providing at least two sets of specimens for synthesizing a reference row, each set comprising at least two groups of cleavage products or a mixture thereof, where the two groups totally contain guanine specific cleavage products, adenine specific cleavage products, thymine specific cleavage products and cytosine specific cleavage products;

(2) providing at least one group of cleavage products or a mixture thereof containing at least one kind of the base specific cleavage products;

(3) resolving both the two sets of specimens provided in step (1) and the product group provided in step (2) one-dimensionally in such parallel relation that at least a first two resolved rows respectively correspond to a first one of said two groups of cleavage products or mixtures thereof and a second two resolved rows respectively correspond to a second one of said two groups of cleavage products or mixtures thereof where said first two resolved rows and said second two resolved rows sandwich at least one resolved row of the product group (2) to form at least five resolved rows on a support medium;

(4) obtaining an autoradiograph having locational information on groups of radioactively labeled cleavage products contained in said rows on the support medium;

(5) generating an electrical digital signal corresponding to said autoradiograph including said row

(6) detecting reference sampling points in each resolved reference row of the digital signal;

(7) detecting sampling points in said one resolved row;

(8) joining corresponding reference sampling points in said reference rows to generate plural continuous lines comprising straight lines, polygonal lines or curved lines; and

(9) identifying said sampling points in said one resolved row of the product group (2) by comparison of the last-mentioned sampling points with said continuous lines to thus determine said base sequence of said DNA or DNA fragment.

(9) causing a stimulable phosphor sheet to absorb radiation and;

(10) scanning said stimulable phosphor sheet with an electromagnetic wave to release at least a portion of radiation energy stored in said phosphor sheet in the form of the autordiograph as stimulated emission, and detecting said stimulated emission photoelectrically.

6. The signal processing method in autoradiography as claimed in claim 5, including resolving on said support resolved rows of at least three sets of said specimens for synthesizing at least three reference where the rows of said first two resolved rows are closely spaced to each other as are the rows of said second two resolved rows.

7. The signal processing method in autoraddiography as claimed in claim 5 or 6, wherein the sampling points are detected by at least smoothing or threshold-processing the digital signal on each scanning line for the resolved rows respectively.

8. The signal processing method in autoradiography as claimed in claim 5 or 6, wherein each set of specimens for synthesizing a reference row consists essentially of two groups of mixtures of base specific cleavage products including:

(1) guanine specific cleavage products and adenine specific cleavage products; and

(2) thymine specific cleavage products and cytosine specific cleavage products.

9. A signal processing method in autoradiography for determination of the base sequence of DNA or a DNA fragment, employing radioactively labeled cleavage products obtained by specifically cleaving the DNA or DNA fragment labeled with a radioactive element, comprising the steps of:

(1) providing at least one set of a reference mixture consisting essentially of guanine specific cleavage products, adenine specific cleavage products, thymine specific cleavage products and cytosine specific cleavage products;

(2) providing at leat one set of a specimen for synthesizing a reference row, the set comprising at least two groups of cleavage products or a mixture thereof, where the two groups in combination totally include guanine specific cleavage products, adenine specific cleavage products, thymine specific cleavage products and cytosine specific cleavage products;

(3) providing at least one group of cleavage products or a mixture thereof containing at least one kind of the base specific cleavage products;

(4) resolving the reference mixture provided in step (1), the specimen set provided in step (2), and the product group provided in step (3) one-dimensionally in such parallel relation that the resolved row of the reference mixture (1) and two or more of resolved rows of the specimen for synthesizing a reference row (2) sandwich at least one resolved row of the product group (3) to form at least four resolved rows on a support medium;

(5) obtaining an autoradiograph having locational information on groups of radioactively labeled cleavage products contained in said rows on the support medium;

(6) detecting sampling points in each resolved row of the digital signal;

(7) assigning both the sampling points in a reference row corresponding to the resolved row of the reference mixture and the sampling points in a reference row synthesized from the two or more of resolved rows of the specimen to reference sampling points;

(8) joining the corresponding reference sampling points in said reference rows to generate plural continuous lines comprising straight lines, polygonal lines or curved lines; and

(9) identifying said sampling points in said one resolved row of the product group (3) by comparison of said last mentioned sampling points with said continuous lines to thus determine said base sequence of said DNA or DNA fragment.

(9) causing a stimulable phosphor sheet to absorb radiation; and

(10) scanning said stimulable phosphor sheet with an electromagnetic wave to release at least a portion of radiation energy stored in said phosphor sheet in the form of the autoradiograph as stimulated emission, and detecting said stimulated emission photoelectrically.

10. The signal processing method in autoradiography as claimed in claim 9, wherein at least three sets of resolved rows including a combination of a resolved row of the reference mixture (1) and two or more resolved rows of the specimen for synthesizing a reference row (2) are so provided on the support medium that said two resolved rows of the specimen are closely spaced and said resolved row of the reference mixture and the reference row (2) sandwich at least one resolved row of the product group (3).

11. The signal processing method in autoradiography as claimed in claim 9 or 10, wherein the sampling points are detected by smoothing or threshold-processing the digital signal on each scanning line for the plural resolved rows respectively.

12. The signal processing method in autoradiography as claimed in claim 9 or 10, wherein the set of specimens for synthesizing reference two (2) consists essentially of two groups of mixtures of base specific cleavage products including:

(1) guanine specific cleavage products and adenine specific cleavage products; and

(2) thymine specific cleavage products and cytosine specific cleavage products.