JPH0621864B2 - Electrophoresis image analysis method - Google Patents
Electrophoresis image analysis methodInfo
- Publication number
- JPH0621864B2 JPH0621864B2 JP59221133A JP22113384A JPH0621864B2 JP H0621864 B2 JPH0621864 B2 JP H0621864B2 JP 59221133 A JP59221133 A JP 59221133A JP 22113384 A JP22113384 A JP 22113384A JP H0621864 B2 JPH0621864 B2 JP H0621864B2
- Authority
- JP
- Japan
- Prior art keywords
- image
- density
- electrophoretic
- shoulder
- electrophoretic image
- 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.)
- Expired - Lifetime
Links
- 238000001962 electrophoresis Methods 0.000 title description 5
- 238000003703 image analysis method Methods 0.000 title 1
- 238000000034 method Methods 0.000 claims description 12
- 238000013508 migration Methods 0.000 claims description 12
- 230000005012 migration Effects 0.000 claims description 12
- 230000003287 optical effect Effects 0.000 description 15
- 102000009027 Albumins Human genes 0.000 description 8
- 108010088751 Albumins Proteins 0.000 description 8
- 238000001514 detection method Methods 0.000 description 7
- 238000010586 diagram Methods 0.000 description 7
- NNBZCPXTIHJBJL-UHFFFAOYSA-N decalin Chemical compound C1CCCC2CCCCC21 NNBZCPXTIHJBJL-UHFFFAOYSA-N 0.000 description 6
- 238000005259 measurement Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- PXXNTAGJWPJAGM-UHFFFAOYSA-N vertaline Natural products C1C2C=3C=C(OC)C(OC)=CC=3OC(C=C3)=CC=C3CCC(=O)OC1CC1N2CCCC1 PXXNTAGJWPJAGM-UHFFFAOYSA-N 0.000 description 3
- 108010044091 Globulins Proteins 0.000 description 2
- 102000006395 Globulins Human genes 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000006734 Beta-Globulins Human genes 0.000 description 1
- 108010087504 Beta-Globulins Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 108010074605 gamma-Globulins Proteins 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
- G01B11/00—Measuring arrangements characterised by the use of optical techniques
- G01B11/02—Measuring arrangements characterised by the use of optical techniques for measuring length, width or thickness
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44717—Arrangements for investigating the separated zones, e.g. localising zones
- G01N27/44721—Arrangements for investigating the separated zones, e.g. localising zones by optical means
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Description
【発明の詳細な説明】 （技術分野） 本発明は、電気泳動像の分析方法に関するものである。TECHNICAL FIELD The present invention relates to a method for analyzing an electrophoretic image.
（従来技術） 従来、蛋白質等の生体高分子を分析する装置として電気
泳動法を利用した自動化学分析装置が実用化されてい
る。この電気泳動法を利用した自動化学分装置では、緩
衝液で湿潤したセルロースアセテート膜等の支持体上に
血清等の検体を塗布し、支持体に通電して電気泳動させ
検体中に含まれる各種成分を分離した泳動像を作り、こ
の泳動像をデンシトメータを用いて光学走査してデンシ
トグラムパターンを造り、このデンシトグラムパターン
を演算処理して検体中に含まれる各種成分を定量分析す
るように構成されている。この自動化学分析装置では、
泳動像と正確に対応したデンシトグラムパターンを作る
ことが要求され、デンシトグラムパターンの形成は分析
精度を高める上で極めて重要である。(Prior Art) Conventionally, an automatic chemical analyzer utilizing an electrophoretic method has been put into practical use as an apparatus for analyzing biopolymers such as proteins. In the automatic chemical fractionation device using this electrophoresis method, a sample such as serum is coated on a support such as a cellulose acetate membrane wetted with a buffer solution, and electricity is applied to the support to cause various components to be electrophoresed. It is configured to perform a densitogram pattern by optically scanning this phoresis image by separating it with a densitometer, and performing a calculation process on this densitogram pattern to quantitatively analyze various components contained in the sample. ing. With this automatic chemical analyzer,
It is required to form a densitogram pattern that exactly corresponds to the electrophoretic image, and the formation of the densitogram pattern is extremely important for improving the analysis accuracy.
第１図Ａは泳動像の一例を示す線図であり、第１図Ｂは
第１図Ａに示す泳動像を光学走査して得られたデンシト
グラムパターンである。泳動像からデンシトグラムパタ
ーンを作るには、電気泳動後に染色及び脱色工程を経た
支持体をデカリン等の透明化液中に浸漬し、浸漬した状
態でデンシトメータを用いて矢印Ｘで示す泳動方向に光
学的に走査し、その透過光から泳動像の各成分の光学的
濃度を測定して第１図Ｂに示すデンシトグラムパターン
を作るように構成されている。デンシトメータで光学走
査するには、まずえ泳動像の泳動方向Ｘと直交するＹ方
向の中心点を検出しなければならず、中心点を正確に検
出しないとデンシトグラムパターンを正確に形成するこ
とができなくなってしまう。FIG. 1A is a diagram showing an example of the electrophoretic image, and FIG. 1B is a densitogram pattern obtained by optically scanning the electrophoretic image shown in FIG. 1A. To create a densitogram pattern from the electrophoretic image, the support that has been subjected to the staining and decolorization process after electrophoresis is immersed in a clearing solution such as decalin, and then in the immersion state, the densitometer is used to optically move in the migration direction indicated by arrow X. Scanning is performed, and the optical density of each component of the electrophoretic image is measured from the transmitted light to form the densitogram pattern shown in FIG. 1B. In order to perform optical scanning with a densitometer, first, the center point in the Y direction orthogonal to the migration direction X of the electrophoretic image must be detected. If the center point is not accurately detected, the densitogram pattern can be accurately formed. I can not do it.
従来の泳動像の中心点を検出する方法として特開昭58-4
5540号公報には、塗布した検体の一番近い位置にできる
アルブミンの第１分画像をデンシトメータでＹ方向に光
学的に走査し、光学濃度が一番高い位置を中心点として
検出する方法が記載されている。しかし、第１分画像の
泳像方向と直交するＹ方向の光学的濃度分布は、第２図
に示すように最大濃度点が中心点とならないパターンが
しばしば発生する。特に第２図のａ及びｂに示すように
肩部の光学濃度が高く中心部の濃度が低くなる泳動像を
生ずる場合が多く、従来の中心点検出方法では泳動像の
中心点を正確に検出することができない欠点があった。
特に、像中心からずれてしまうと光ビームのスリットが
斜行してしまい泳動像からずれた位置を光学走査する不
都合が生ずるため、像中心を正確に決定できる電気泳動
像の分析方法の開発が要請されている。As a conventional method for detecting the center point of an electrophoretic image, JP-A-58-4
5540 discloses a method of optically scanning the first minute image of albumin formed at the position closest to the applied sample in the Y direction with a densitometer and detecting the position with the highest optical density as the center point. Has been done. However, the optical density distribution in the Y direction orthogonal to the swimming image direction of the first minute image often has a pattern in which the maximum density point is not the center point as shown in FIG. In particular, as shown in FIGS. 2A and 2B, a migration image in which the optical density of the shoulder portion is high and the density of the center portion is low is often generated, and the conventional center point detection method accurately detects the center point of the migration image. There was a drawback that could not be done.
In particular, when the light beam is displaced from the center of the image, the slit of the light beam is skewed, which causes a disadvantage of optically scanning a position displaced from the electrophoresis image. Has been requested.
（発明の目的） 本発明の目的は上述した欠点を除去し、泳動像の中心点
を正確に決定できる電気泳動像の分析方法を提供するも
のである。(Object of the Invention) An object of the present invention is to eliminate the above-mentioned drawbacks and to provide a method for analyzing an electrophoretic image capable of accurately determining the center point of the electrophoretic image.
（発明の概要） 本発明による電気泳動像の分析方法は、支持体上に形成
された検体の泳動像を、この検体中に含まれる各成分の
分画像が順次形成されている泳動方向に沿って光学的に
走査して検体のデンシトグラムパターンを検出するに際
し、分析すべき検体中に含まれる１の成分の分画像につ
いて、泳動方向と直交する方向に沿って光学的に走査し
て支持体上の泳動像が存在していない領域と肩部との間
の濃度プロフィールを求め、求めた濃度プロフィール上
の、泳動像が存在していない領域の濃度Ｄ０から、肩部
濃度ＤＨと前記泳動像が存在していない領域の濃度Ｄ０
との間の差の所定の比率だけ増大した特定点を求め、前
記特定点から泳動像の中心に向けて所定の距離だけ離間
した位置に対応する泳動像上の位置を泳動像の中心位置
とし、この中心位置を通るように泳動方向に沿って光学
的に走査して検体中に含まれる各成分のデンシトグラム
パターンを検出することを特徴とするものである。(Summary of the Invention) A method for analyzing an electrophoretic image according to the present invention is a method in which an electrophoretic image of a sample formed on a support is taken along the electrophoretic direction in which partial images of respective components contained in the sample are sequentially formed. When the densitogram pattern of the sample is detected by optically scanning with a support, the sub-image of one component contained in the sample to be analyzed is optically scanned along the direction orthogonal to the migration direction. The concentration profile between the region where the electrophoretic image does not exist and the shoulder is obtained, and the concentration D H of the shoulder region is calculated from the concentration D 0 of the region where the electrophoretic image does not exist on the obtained concentration profile. Density D 0 in the area where no electrophoretic image exists
The specific point increased by a predetermined ratio of the difference between and, the position on the electrophoretic image corresponding to the position separated by a predetermined distance from the specific point toward the center of the electrophoretic image is the center position of the electrophoretic image. The densitogram pattern of each component contained in the sample is detected by optically scanning along the migration direction so as to pass through the center position.
（実施例） 第３図は本発明による電気泳動像の分析方法を実施する
ための比色定量装置の一例の構成を示す線図的断面図で
ある。染色及び脱色工程を経た泳動像が形成されている
細条又はシート状の支持体１を、透明化液を満した透明
容器２内に搬入する。透明容器２の入口部には一対の供
給ローラ３及び４を配置し、支持体１はこの供給ローラ
３及び４により透明容器２内に搬入され、まず、光源５
と光検出器６からなる先端検出センサによりその先端部
が検出される。先端部の検出後透明化液であるデカリン
液内に浸入する。この透明化液は支持体１だけを透明化
するものであり、デカリン内に浸入した支持体は、透明
化された支持体上に泳動像が形成された状態となる。次
に、第１の搬送ローラ７及び８により透明容器２の底部
に形成された測定領域に搬入され、泳動像の光学濃度が
測定される。測定後第２の搬送ローラ９及び10により挟
持搬送され、排出ローラ11及び12を経て外部に排出され
る。測定領域には、透明容器２の下側に光源13、熱線吸
収フィルタ14、レンズ15、フィルタ16、直角プリズム17
及びスリット18から成る光源装置を設けると共に、透明
容器２の上側には受光器19を設け、この受光器19を比色
計20に接続する。光源13から発した光ビームはレンズ15
及びスリット18を経てスポット状に収束されて支持体１
を透過し受光体19で受光され、泳動像の光学濃度が比色
計20により検出される。光源装置及び受光器19は、駆動
装置（図示せず）に一体的に連結され各検体の分画像が
順次形成されている支持体１の幅方向、すなわち紙面に
垂直方向に移動して支持体上に形成されている泳動像を
その泳動方向に光学的に走査する。また、第２の搬送ロ
ーラ９及び10は、パルスモータ（図示せず）に連結され
支持体１を１ステップずつ間欠的に搬送する。そして、
第２の搬送ローラ９及び10が１ピッチ移動する毎に光源
装置及び受光器19を支持体１の幅方向に移動させて光学
走査を行なうように構成する。(Example) FIG. 3 is a schematic cross-sectional view showing the configuration of an example of a colorimetric apparatus for carrying out the method for analyzing an electrophoretic image according to the present invention. The strip-shaped or sheet-shaped support 1 on which the electrophoretic image formed through the dyeing and decoloring steps is formed is carried into a transparent container 2 filled with a clearing solution. A pair of supply rollers 3 and 4 are arranged at the entrance of the transparent container 2, and the support 1 is carried into the transparent container 2 by the supply rollers 3 and 4, and first, the light source 5
The tip portion is detected by the tip detection sensor including the photodetector 6. After the tip is detected, it penetrates into the decalin solution, which is the clearing solution. This clearing liquid makes only the support 1 transparent, and the support that has penetrated into decalin is in a state in which a migration image is formed on the transparent support. Next, the first transport rollers 7 and 8 carry the sample into the measurement area formed on the bottom of the transparent container 2, and the optical density of the electrophoretic image is measured. After the measurement, the sheet is nipped and conveyed by the second conveying rollers 9 and 10, and is discharged to the outside via the discharging rollers 11 and 12. In the measurement area, a light source 13, a heat ray absorbing filter 14, a lens 15, a filter 16 and a right angle prism 17 are provided below the transparent container 2.
A light source device including a slit and a slit 18 is provided, and a light receiver 19 is provided on the upper side of the transparent container 2, and the light receiver 19 is connected to a colorimeter 20. The light beam emitted from the light source 13 is a lens 15
And the support 1 after being converged in a spot shape through the slit 18.
And is received by the light receiver 19, and the optical density of the electrophoretic image is detected by the colorimeter 20. The light source device and the light receiver 19 are moved integrally in the width direction of the support body 1 which is integrally connected to a driving device (not shown) and in which partial images of each sample are sequentially formed, that is, in the direction perpendicular to the paper surface. The electrophoretic image formed above is optically scanned in the electrophoretic direction. The second transport rollers 9 and 10 are connected to a pulse motor (not shown) and intermittently transport the support 1 step by step. And
Each time the second conveying rollers 9 and 10 move by one pitch, the light source device and the light receiver 19 are moved in the width direction of the support 1 to perform optical scanning.
第４図は光源装置から発する光ビームの支持体上の軌跡
を示す線図である。支持体１上には、支持体の搬送方向
であるＹ方向に順次各検体の泳動像が形成され、Ｘ方向
には検体の各成分の分画像が順次形成されている。本例
では検体として血清を用い血清中に含まれる各種蛋白質
を定量分析するものとし、第１分画像としてアルブミン
の泳動像が形成され、泳動方向であるＸ方向に順次α１
グロブリン，α２グロブリン，βグロブリン，γグロブ
リンの分画像が形成されている。本例では光学走査によ
り第１分画像であるアルブミンの分画像から電気泳動像
の中心点を求める。支持体１は第２搬送ローラ９及び10
により１ステップずつ間欠的にＹ方向に搬送され、光ビ
ームは１ステップ移動する毎にアルブミンの泳動像をほ
ぼ越える位置ます移動する。これによりアルブミンの泳
動像はＸ及びＹ方向に２次元的に走査されることにな
る。そして、光ビームのＸ方向の走査における光学濃度
の最大値を用いて濃度プロフィールを作る。このように
して得られたアルブミンの濃度プロフィールを第５図に
示す。第５図は３個の検体について連続的に測定したも
のであり、アルブミンの泳動像のプロフィールと正確に
対応している。FIG. 4 is a diagram showing the trajectory of the light beam emitted from the light source device on the support. On the support 1, electrophoretic images of each sample are sequentially formed in the Y direction that is the transport direction of the support, and partial images of each component of the sample are sequentially formed in the X direction. In this example, it is assumed that serum is used as a sample and various proteins contained in the serum are quantitatively analyzed, and an electrophoretic image of albumin is formed as a first minute image, and α 1 is sequentially transferred in the X direction, which is the electrophoretic direction.
Separate images of globulin, α 2 globulin, β globulin, and γ globulin are formed. In this example, the center point of the electrophoretic image is obtained from the minute image of albumin, which is the first minute image, by optical scanning. The support 1 includes the second transport rollers 9 and 10
Thus, the light beam is intermittently transported in the Y direction step by step, and the light beam moves to a position almost exceeding the electrophoretic image of albumin every time it moves one step. As a result, the electrophoretic image of albumin is two-dimensionally scanned in the X and Y directions. Then, a density profile is created using the maximum value of the optical density in the scanning of the light beam in the X direction. The concentration profile of albumin thus obtained is shown in FIG. FIG. 5 shows continuous measurement of three specimens, which accurately corresponds to the profile of the electrophoretic image of albumin.
各検体の濃度プロフィールは、濃度がきわめて低い像間
部ａから発し、濃度が直線的に且つ急激に増加する前縁
部ｂに移行し、肩部ｃを経て中央部ｄに達している。濃
度プロフィールの形態について種々の検討を加えた結
果、次のような特徴を有することが確認された。The concentration profile of each sample starts from the inter-image portion a where the concentration is extremely low, moves to the front edge portion b where the concentration increases linearly and sharply, and reaches the central portion d via the shoulder portion c. As a result of various studies on the form of the concentration profile, it was confirmed that it has the following features.
(1)像間部ａの濃度は、光を反射するスリット18が有限
の幅を有しているため、各検体間において相異してい
る。(1) The density of the inter-image portion a is different among the samples because the slit 18 that reflects light has a finite width.
(2)前縁部ｂは直線的に濃度が増加しており、かつその
勾配は急峻であり、各検体間において一致している。(2) The concentration at the front edge portion b increases linearly, and the gradient is steep, which is the same in each specimen.
(3)肩部ｃではゆるやかに濃度が増加しているため、そ
の発生位置の特定は若干誤差を生ずるおそれがある。(3) Since the density of the shoulder c is gradually increasing, there is a possibility that a slight error will occur in the specification of the position where the density occurs.
(4)肩部間の幅ｌ１は、各検体間において若干相異して
いるだけであり、ほぼ一致している。(4) The width l 1 between the shoulders is slightly different between the samples, and is almost the same.
(5)像中心点を通る中央部ｄの濃度は、各検体間におい
て相異し、最大濃度点は像中心と必らずしも一致してい
ない。(5) The densities of the central portion d passing through the image center point differ among the samples, and the maximum density point does not necessarily coincide with the image center.
以上の結果より、像中心を検出するに当り、次の結論が
得られる。From the above results, the following conclusions can be obtained in detecting the image center.
(1)像間部ａの立ち上がり点及び肩部の位置付近では、
位置に対して濃度がゆるやかに変化しているため、像間
部又は肩部の位置を基準にして像中心を求めると、誤差
が生じ易すい。(1) In the vicinity of the rising point of the inter-image part a and the position of the shoulder,
Since the density changes gently with respect to the position, if the image center is obtained with the position of the inter-image portion or the shoulder portion as a reference, an error easily occurs.
(2)前縁部では濃度が急激に増加しているから、濃度変
化に対する位置の変化は極めて小さい。しかも前縁部ｂ
の濃度勾配は各検体間において肩部濃度に対応して大き
くなっている。(2) Since the density at the front edge increases rapidly, the change in position with respect to the density change is extremely small. Moreover, the front edge b
The concentration gradient of is large between the samples in accordance with the shoulder concentration.
従って、前縁部ｂ上の特定濃度点を定め、この特定濃度
点を位置に所定の距離を加えれば各検体の濃度プロフィ
ールから誤差を生ずることなく像中心が求まる。Therefore, by defining a specific density point on the front edge portion b and adding a predetermined distance to the position of this specific density point, the image center can be obtained from the density profile of each sample without causing an error.
以上の考察より本発明では肩部ｃの濃度及び検体間に位
置する像間の濃度プロフィールを検出し、この像間濃度
と肩部の濃度との間の差に対して所定の割合にある前縁
部ｂ上の特定濃度点を定め、この特定濃度点の位置を基
準にして予め求めた所定の距離ｄ０を加えた点の位置を
求め、これを像中心とする。From the above consideration, according to the present invention, the density of the shoulder c and the density profile between the images located between the samples are detected, and the difference between the density between the images and the density of the shoulder is within a predetermined ratio. A specific density point on the edge b is determined, a position of a point to which a predetermined distance d 0 calculated in advance is added on the basis of the position of the specific density point is determined, and this is used as the image center.
次に本発明による像中心検出プロセスを第４図及び第６
図を参照しながら説明する。尚、第６図は光学走査によ
り得られた濃度プロフィールパターン図である。第２搬
送ローラ９及び10により、支持体１をステップ送りし、
これと同期して光源装置及び受光器19を支持体１の幅方
向に移動させて光学走査を行ない、第１分画像であるア
ルブミンの泳動像方向と直交する方向の濃度プロフィー
ルを検出する。濃度プロフィールの作成は、第２搬送ロ
ーラのステップ送り量から位置を求め、その位置及びそ
の位置で検出した光学濃度をそれぞれ記憶して行なう。
まず、検体と検体との中間の位置を基準位置Ｓ０として
定め、かつ支持体の先頭で検体がない基準位置S0の光学
濃度D0 を検出する。各検体の泳動像は、ほぼ所定のピ
ッチ間隔で形成されているから、支持体１の先端検出操
作から正確に定めることができる。Next, the image center detection process according to the present invention will be described with reference to FIGS.
Description will be given with reference to the drawings. Incidentally, FIG. 6 is a density profile pattern diagram obtained by optical scanning. The support 1 is stepwise fed by the second transport rollers 9 and 10,
In synchronization with this, the light source device and the light receiver 19 are moved in the width direction of the support 1 to perform optical scanning, and the concentration profile in the direction orthogonal to the electrophoretic image direction of albumin, which is the first minute image, is detected. The density profile is created by obtaining the position from the step feed amount of the second transport roller and storing the position and the optical density detected at that position.
First, an intermediate position between samples is set as the reference position S 0 , and the optical density D 0 at the reference position S 0 where there is no sample at the head of the support is detected. Since the electrophoretic image of each sample is formed at a substantially predetermined pitch interval, it can be accurately determined from the operation of detecting the tip of the support 1.
次に基準位置S0からステップ送りと同期して濃度検出を
行なう。像間部及び前縁部の検出では、次式に従う検出
プロセスを行なう。Next, density detection is performed from the reference position S 0 in synchronization with step feed. In detecting the inter-image portion and the leading edge portion, a detection process according to the following equation is performed.
Ｄｎ−Ｄn-1＞Ｃ１(n＝0,1…)…(1) ここで、Ｄｎはｎ番目に検出した濃度を示し、Ｄn-1は
n-1番目に検出した濃度を示す。Ｃ１は基準濃度差であ
り、各検体の前縁部の勾配に基き適切な値を実験により
予め求めておく。像間部では濃度変化が小さく、一方前
縁部は急激な濃度増加を呈するから，直前に検出した濃
度との濃度差がＣ１より大きければ前縁部を検出してい
ることになる。 D n -D n-1> C 1 (n = 0,1 ...) ... (1) where, D n denotes the concentration detected in the n-th, D n-1 is
The n-1st detected concentration is shown. C 1 is a reference concentration difference, and an appropriate value is experimentally obtained in advance based on the gradient of the leading edge of each sample. Since the density change is small in the inter-image area and the leading edge shows a sharp increase in density, if the density difference from the density detected immediately before is larger than C 1 , the leading edge is detected.
次に肩部の位置及び濃度を検出する。肩部の位置検出は
次式に基き行なう。Next, the position and density of the shoulder are detected. The shoulder position is detected based on the following equation.
Ｄｎ−Ｄn-1＜C2 Ｄn-1−Ｄn-2＜C2 …(2) 肩部は急激な濃度増加からゆるやかな増加に移行する点
であるから、直前に検出した濃度との濃度差が所定の値
C2よりも小さくなる点が肩部となる。このC2の値も実験
により適切な値を求めておく。本例では(2)式に基き肩
部の位置S1を検出し、この肩部の濃度ＤＨも検出する。
そして、検出し記憶されている前縁部の濃度プロフィー
ルから、肩部の濃度ＤＨと基準点濃度Ｄ０との濃度差の
半分の濃度に対応する位置S2を検出する。次に、検出し
た位置S2に距離ｄ０に相当するパルス数を加え、対応す
る位置S3を求めて像中心とする。このばあい加える距離
d0は、複数の検体について計測した値を統計処理して予
め求めて入力しておく。次に、肩部の位置S1と像中心S3
とのステップ差を検出し、このステップ数だけ移行し、
像中心に到達した時、支持体１の全幅まで光学走査し
て、デンシトグラムパターンを作る。デンシトグラムパ
ターンを検出した後、像間部までの所定のステップ数だ
け自動的に送り、次の検出に具えるこのように、肩部の
位置及び濃度から基準となる濃度点を求める構成とすれ
ば中央部まで光学走査することなく像中心を検出でき、
走査時間を短縮できる効果もある。D n -D n-1 <C 2 D n-1 -D n-2 <C 2 (2) Since the shoulder part is a point where the concentration increases suddenly and gradually increases, the concentration detected immediately before The density difference with
The point that is smaller than C 2 is the shoulder. An appropriate value for this C 2 value is also obtained by experiments. In this example, the position S 1 of the shoulder is detected based on the equation (2), and the density D H of this shoulder is also detected.
Then, from the detected and stored density profile of the front edge portion, the position S 2 corresponding to the half density of the density difference between the shoulder density DH and the reference point density D 0 is detected. Next, the number of pulses corresponding to the distance d 0 is added to the detected position S 2 , and the corresponding position S 3 is obtained and set as the image center. In this case, the distance to add
For d 0 , the values measured for a plurality of specimens are statistically processed and obtained in advance and input. Next, the shoulder position S 1 and the image center S 3
Detects the step difference between and, moves by this number of steps,
When it reaches the center of the image, it is optically scanned over the full width of the support 1 to create a densitogram pattern. After detecting the densitogram pattern, it is automatically sent by a predetermined number of steps up to the inter-image area, and it is ready for the next detection. For example, the center of the image can be detected without optical scanning to the center.
There is also an effect that the scanning time can be shortened.
本発明は上述した実施例にのみ限定されるものではなく
幾多の変更や変形が可能である。例えば上述した実施例
では、肩部の濃度ＤＨと基準の濃度Ｄ０との濃度差が1/
2 になる位置を基準としたが、肩部と像間部との間の前
縁部であれば他の任意の比率の点を基準とすることがで
き、濃度が1/2になる点に限定されるものではない。The present invention is not limited to the above-described embodiments, but various modifications and variations are possible. For example, in the above-described embodiment, the density difference between the shoulder density D H and the reference density D 0 is 1 /
Although the position of 2 was used as the standard, the point of any other ratio can be used as the standard if it is the front edge between the shoulder and the inter-image area, and It is not limited.
（発明の効果） 以上説明したように本発明によれば、支持体上の泳動像
が存在しない領域から肩部までの濃度プロフィールを検
出し、これに基いて像中心を求めるようにしたため、像
中心の濃度がどのようなものであっても像中心を常に正
確に検出することができる。さらに、泳動像中の一部の
領域だけを光学走査するだけで像中心を迅速かつ正確に
検出することができる。(Effect of the invention) As described above, according to the present invention, the density profile from the region where the electrophoretic image on the support does not exist to the shoulder is detected, and the image center is determined based on the density profile. The image center can always be detected accurately regardless of the density of the center. Furthermore, the image center can be detected quickly and accurately only by optically scanning only a part of the electrophoretic image.
第１図Ａは泳動像の一例を示す線図、 同図Ｂは泳動像を光学走査して得られたデンシトグラム
パターン、 第２図は第１分画像の濃度分布を示すグラフ、 第３図は本発明による像中心検出方法を実施するための
比色定量装置の一例の構成を示す線図的断面図、 第４図は光源装置から発する光ビームの支持体上の軌跡
を示す線図、 第５図は各種検体の泳動方向と直交する方向の濃度プロ
フィールを示す線図、 第６図は一検体分の濃度プロフィールを示す線図であ
る。 １……支持体、２……透明容器 ３，４……供給ローラ、５，１３……光源 ６……光検出器、７，８……第１搬送ローラ ９，10……第２搬送ローラ 11,13……排出ローラ、14……熱線吸収フィルタ 15……レンズ、16……フィルタ 17……直角プリズム、18……スリット 19……受光器、20……比色計FIG. 1A is a diagram showing an example of the electrophoretic image, FIG. 1B is a densitogram pattern obtained by optically scanning the electrophoretic image, FIG. 2 is a graph showing the density distribution of the first minute image, and FIG. FIG. 4 is a schematic cross-sectional view showing the configuration of an example of a colorimetric quantification device for carrying out the image center detection method according to the present invention, and FIG. 4 is a diagram showing the trajectory of a light beam emitted from a light source device on a support, FIG. 5 is a diagram showing concentration profiles of various samples in a direction orthogonal to the migration direction, and FIG. 6 is a diagram showing concentration profiles of one sample. 1 ... Support body, 2 ... Transparent container 3,4 ... Supply roller, 5, 13 ... Light source 6 ... Photodetector, 7, 8 ... First transport roller 9, 10 ... Second transport roller 11, 13 ...... Ejection roller, 14 ...... Heat ray absorption filter 15 …… Lens, 16 …… Filter 17 …… Right angle prism, 18 …… Slit 19 …… Photoreceiver, 20 …… Colorimeter
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 昭57−204437（ＪＰ，Ａ) 特開 昭58−45540（ＪＰ，Ａ) 特公 昭47−44680（ＪＰ，Ｂ１) ─────────────────────────────────────────────────── ─── Continuation of the front page (56) Reference JP-A-57-204437 (JP, A) JP-A-58-45540 (JP, A) JP-B 47-44680 (JP, B1)
Claims (1)
の検体中に含まれる各成分の分画像が順次形成されてい
る泳動方向に沿って光学的に走査して検体のデンシトグ
ラムパターンを検出するに際し、 分析すべき検体中に含まれる１の成分の分画像につい
て、泳動方向と直交する方向に沿って光学的に走査して
支持体上の泳動像が存在していない領域と肩部との間の
濃度プロフィールを求め、 求めた濃度プロフィール上の、泳動像が存在していない
領域の濃度Ｄ０から、肩部濃度ＤＨと前記泳動像が存在
していない領域の濃度Ｄ０との間の差の所定の比率だけ
増大した特定点を求め、 前記特定点から泳動像の中心に向けて所定の距離だけ離
間した位置に対応する泳動像上の位置を泳動像の中心位
置とし、 この中心位置を通るように泳動方向に沿って光学的に走
査して検体中に含まれる各成分のデンシトグラムパター
ンを検出することを特徴とする電気泳動像の分析方法。1. A densitogram of a specimen by optically scanning a migration image of the specimen formed on a support along the migration direction in which partial images of each component contained in the specimen are sequentially formed. When detecting a pattern, a partial image of one component contained in the sample to be analyzed is optically scanned along a direction orthogonal to the migration direction and a region where the migration image on the support does not exist is detected. The density profile between the shoulder and the shoulder is calculated, and from the density D 0 of the area where the electrophoretic image does not exist on the obtained density profile, the shoulder density DH and the density D of the area where the electrophoretic image does not exist. The specific point increased by a predetermined ratio of the difference between 0 and 0 is obtained, and the position on the electrophoretic image corresponding to the position separated from the specific point by the predetermined distance toward the center of the electrophoretic image is the center position of the electrophoretic image. And run it so that it passes through this center position A method for analyzing an electrophoretic image, which comprises optically scanning along a direction to detect a densitogram pattern of each component contained in a sample.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59221133A JPH0621864B2 (en) | 1984-10-23 | 1984-10-23 | Electrophoresis image analysis method |
| DE19853537199 DE3537199A1 (en) | 1984-10-23 | 1985-10-18 | Determining mid-point of electrophoretic image - by adding specified distance to coordinate of mean point of leading edge |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59221133A JPH0621864B2 (en) | 1984-10-23 | 1984-10-23 | Electrophoresis image analysis method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6199841A JPS6199841A (en) | 1986-05-17 |
| JPH0621864B2 true JPH0621864B2 (en) | 1994-03-23 |
Family
ID=16761970
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59221133A Expired - Lifetime JPH0621864B2 (en) | 1984-10-23 | 1984-10-23 | Electrophoresis image analysis method |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JPH0621864B2 (en) |
| DE (1) | DE3537199A1 (en) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57204437A (en) * | 1981-06-11 | 1982-12-15 | Hiranuma Sangyo Kk | Measuring method for interval of inspection body in concentration measuring apparatus |
| JPS5845540A (en) * | 1981-09-14 | 1983-03-16 | Toyo Kagaku Sangyo Kk | Detecting method for central part of inspection body in densitometer |
-
1984
- 1984-10-23 JP JP59221133A patent/JPH0621864B2/en not_active Expired - Lifetime
-
1985
- 1985-10-18 DE DE19853537199 patent/DE3537199A1/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| DE3537199C2 (en) | 1989-03-30 |
| DE3537199A1 (en) | 1986-04-24 |
| JPS6199841A (en) | 1986-05-17 |
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