JPS63282653A - Method for measuring activity of cell - Google Patents
Method for measuring activity of cellInfo
- Publication number
- JPS63282653A JPS63282653A JP11779887A JP11779887A JPS63282653A JP S63282653 A JPS63282653 A JP S63282653A JP 11779887 A JP11779887 A JP 11779887A JP 11779887 A JP11779887 A JP 11779887A JP S63282653 A JPS63282653 A JP S63282653A
- Authority
- JP
- Japan
- Prior art keywords
- image
- fertilized egg
- change
- cell
- activity
- 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.)
- Pending
Links
- 230000000694 effects Effects 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims description 17
- 238000003384 imaging method Methods 0.000 claims description 7
- 230000008859 change Effects 0.000 abstract description 13
- 238000012545 processing Methods 0.000 abstract description 13
- 239000007788 liquid Substances 0.000 abstract 1
- 235000013601 eggs Nutrition 0.000 description 22
- 210000004027 cell Anatomy 0.000 description 12
- 238000005259 measurement Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000000691 measurement method Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 230000035935 pregnancy Effects 0.000 description 2
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- BPPVUXSMLBXYGG-UHFFFAOYSA-N 4-[3-(4,5-dihydro-1,2-oxazol-3-yl)-2-methyl-4-methylsulfonylbenzoyl]-2-methyl-1h-pyrazol-3-one Chemical compound CC1=C(C(=O)C=2C(N(C)NC=2)=O)C=CC(S(C)(=O)=O)=C1C1=NOCC1 BPPVUXSMLBXYGG-UHFFFAOYSA-N 0.000 description 1
- 241001091551 Clio Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 239000003761 preservation solution Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
【発明の詳細な説明】
［発明の１］的］
（産業上の利用分野）
本発明は、細胞の活性度測定方法に係り、特に受精卵等
の活性度の測定に好適な細胞の活性度測定方法に関する
。Detailed Description of the Invention [Objective of the Invention 1] (Industrial Application Field) The present invention relates to a method for measuring the activity of cells, and in particular, a method for measuring the activity of cells suitable for measuring the activity of fertilized eggs, etc. Regarding measurement methods.
（従来の技術）
近年は、哺乳動物の体内から採取した、受精卵や未受精
卵子に体外受精した受精卵を子宮内に移植して発育させ
るいわゆる人工妊娠の技術が急速に進歩している。(Prior Art) In recent years, there has been rapid progress in so-called artificial pregnancy technology, in which a fertilized egg or an unfertilized egg collected from a mammal's body is in vitro fertilized and implanted into the uterus for development.
このような人工妊娠においては、受精卵か移植に適する
かどうか検査する必要がある。また移植を行って受胎す
る可能性があると判定された場合どの程度の受胎成績が
期待できる受精卵なのか品質も検査する。従来は、顕Ｖ
＆鏡下で形態を観察し、一定の基準に照らし合わせて受
精卵の品質を判定する方法が広く行われている。この方
法は、受精卵の形態観察の経験がある程度要求され、か
なり主観的な方法であり、受胎率の向上を阻害する要因
となっている。In such artificial pregnancies, it is necessary to test whether the fertilized eggs are suitable for implantation. In addition, the quality of the fertilized eggs is also inspected to see what level of fertilization results can be expected if it is determined that there is a possibility of conception after transplantation. Conventionally, the
& A widely used method is to observe the morphology under a mirror and judge the quality of the fertilized eggs based on certain criteria. This method requires a certain degree of experience in observing the morphology of fertilized eggs, and is a highly subjective method, which is a factor that hinders the improvement of the conception rate.
（発明が解決しようとする問題点）
しかしながら、受精卵に生じる変化の速度は遅く、上述
のように、顕微鏡により目視する方法では、受精卵に生
じる変化の度合、すなわち活性度の違いを正確に知るこ
とができなかった。(Problem to be solved by the invention) However, the rate of change that occurs in the fertilized egg is slow, and as mentioned above, the method of visual observation using a microscope cannot accurately determine the degree of change that occurs in the fertilized egg, that is, the difference in activity. I couldn't know.
本発明は、かかる従来の事情に対処してなされたもので
、受精卵等の動きおよび変化の速度の遅いＭＡｌｉ２！
でも、その活性度を正確に測定することのできる細胞の
活性度測定方法を提供しようとするものである。The present invention has been made in response to such conventional circumstances, and the MAli2!
However, the present invention aims to provide a method for measuring cell activity that can accurately measure the activity.
［発明の構成コ
（問題点を解決するための手段）
すなわち本発明の細胞の活性度測定方法は、細胞の拡大
像を撮像手段により電気的な画像信号に変換し、この画
像信り−を画像データとして記憶１段に記憶させ、この
後所定時間経過後の前記細胞の画像信号を前記撮像手段
から入力し前記記憶手段に記憶された前記画像データと
比較することを特徴とする。[Configuration of the Invention (Means for Solving Problems) In other words, the cell activity measurement method of the present invention converts an enlarged image of a cell into an electrical image signal by an imaging means, and converts the image reliability into an electrical image signal. It is characterized in that it is stored as image data in a first storage stage, and then an image signal of the cells after a predetermined period of time is inputted from the imaging means and compared with the image data stored in the storage means.
（作 用）
本発明の細胞の活性度測定方法では、細胞の拡大像を撮
像手段により電気的な画像借りに変換し、この画像信号
を画像データとして記憶手段に記憶させ、この後所定時
間経過後の細胞の画像信号を撮像手段から入力し、記憶
手段に記憶された画像データと例えばデジタル減算処理
による画像演算等を行うことにより比較する。(Function) In the cell activity measuring method of the present invention, an enlarged image of a cell is converted into an electrical image by an imaging means, this image signal is stored as image data in a storage means, and then a predetermined period of time has elapsed. A subsequent image signal of the cell is input from the imaging means and compared with the image data stored in the storage means by performing image calculations such as digital subtraction processing.
したがって、受精卵等の動きおよび変化の速度の遅い細
胞でも、その活性度を正確にＭ１定することができる。Therefore, even in cells that move and change slowly, such as a fertilized egg, the activity level M1 can be determined accurately.
（実施例）
以下本発明の活性度測定方法を図面を参照して実施例に
ついて説明する。(Example) Examples of the activity measuring method of the present invention will be described below with reference to the drawings.
第１図は、この実施例方法に使用する装置の構成を示す
もので、倒立ｍｓ鏡１と、倒立顕微鏡１の像を撮像する
ためのＣＣＤカメラ２等の撮像手段と、ＣＣＤカメラ２
によって得られた画像信号を処理する画像処理装置３と
から構成されている。FIG. 1 shows the configuration of an apparatus used in this embodiment method, which includes an inverted MS mirror 1, an imaging means such as a CCD camera 2 for capturing an image of the inverted microscope 1, and a CCD camera 2.
and an image processing device 3 that processes the image signal obtained by.
なお、画像処理＊ ’ｒａ ３は、Ａ／Ｄ変換器３ａ、
グラフィックプロセッサ３ｂ、メモリ３Ｃ等から構成さ
れている。Note that image processing *'ra 3 is an A/D converter 3a,
It is composed of a graphic processor 3b, a memory 3C, and the like.
上記構成の装置を用いてこの実施例では、次のようにし
て受精卵の活性度を測定する。In this example, the activity of a fertilized egg is measured in the following manner using the apparatus configured as described above.
まず、保存液等に浸した受精卵が配置されたシャーレ１
１を倒立顕微鏡１の測定ステージに配置し、マイクロマ
ニピュレータ１２等でこの受精卵が移動しないように保
持してサンプルセットを行う、なお、倒立顕微鏡１の倍
率は、例えば４０倍程度とし、上部から照明１３による
照明を行う。First, a Petri dish 1 containing fertilized eggs soaked in a preservation solution, etc.
1 is placed on the measurement stage of the inverted microscope 1, and the sample is set by holding the fertilized egg using a micromanipulator 12 or the like so that it does not move.The magnification of the inverted microscope 1 is, for example, about 40x, and the sample is set from the top. Illumination is performed by illumination 13.
次に、ＣＣＤカメラ２のフォーカス調整を手動または自
動で行い、画像処理装置３による処理を開始する。Next, focus adjustment of the CCD camera 2 is performed manually or automatically, and processing by the image processing device 3 is started.
画像処理装置３は、第２図のフＩ７−チャートにも示す
ように、次のような処理を行う。The image processing device 3 performs the following processing, as shown in the chart I7 of FIG.
すなわち、まずＡ／Ｄ変換器３ａのゲイン調整を行ない
（ａ）ｃｌＩＯ的に、サンプルの位置すなわち計測位置
を検出する（ｂ）。That is, first, the gain of the A/D converter 3a is adjusted (a) and the position of the sample, that is, the measurement position is detected using CLIO (b).
次に、サンプルの画像をマスター画像としてグラフィッ
クプロセッサ３ｂに入力しくＣ）、メモリ３ｃ内に記憶
する（ｄ）。Next, the sample image is input as a master image to the graphic processor 3b (c) and stored in the memory 3c (d).
この後、Ｎ＝ １にカウント設定を行い（ｅ）、あらか
じめ設定された所定時間例えば数秒ないし数１−秒待伏
する（ｆ）。Thereafter, a count is set to N=1 (e), and the process waits for a preset predetermined time, for example, several seconds to several seconds (f).
上記所定時間経過後、再びグラフィック１０センサ３ｂ
に、Ｎ・・１のサンプルの画像を人力しくｇ）、メモリ
３Ｃ内に記憶されたマスター画像とデジタル減算処理に
よる画像演算を行う（１１）。After the predetermined time has elapsed, the graphic 10 sensor 3b
Next, the images of the N...1 samples are manually processed (g) and image calculations are performed by digital subtraction processing with the master image stored in the memory 3C (11).
次に、上記演３ｔＶ１里からヒストグラムを算出しくｉ
）、カウントをＮ＋１とする（ｊ）。Next, calculate the histogram from the above performance 3tV1ri.
), and the count is set to N+1 (j).
そして、」−記カウントと、予め設定された測定回数Ｍ
と比較し、Ｎ＜Ｍの場合は上述の（ｆ）以後の処理を繰
り返して行い、Ｎ＜Ｍでない場合は次の処理に進む（ｋ
）。and the count and the preset number of measurements M.
If N<M, the process from (f) above is repeated, and if N<M, proceed to the next process (k
).
上記比較結果がＮ＜Ｍでない場合は、処理（ｉ）で得ら
れたヒストグラムの配列データから活性度を算出しくフ
）、この算出結果を出力して（ｍ）、終了する＜ｎ）。If the above comparison result is not N<M, calculate the activity level from the histogram array data obtained in process (i) (f), output this calculation result (m), and end <n).
すなわち、まず、受精卵の画像（１４ＡＳＴＬＲ１をマ
スター画像として入力して記憶し、この後、所定時間経
過後の受精卵の画像（ＤＬＳＴＩＮ＾Ｔｌ０Ｎ）を入力
し、マスター画像とのデジタル減算処理による画像演算
を行う。That is, first, the image of the fertilized egg (14ASTLR1) is input and stored as a master image, then the image of the fertilized egg after a predetermined time (DLSTIN^Tl0N) is input, and the image is created by digital subtraction processing with the master image. Perform calculations.
上記画像演算の結果得られる画像は、全く変１ヒのない
部分については、真黒の画像となり、変化があった部分
の画像については、その変化の度合により、次第に明る
くなる。このようなグレイレベル毎にビクセル（画素）
の数をカランｌ−Ｌ、第３図に示すようなグレイレベル
毎のヒストグラムを作成する。The image obtained as a result of the above-mentioned image calculation will be a pure black image for areas where there is no change at all, and the image for areas where there has been a change will gradually become brighter depending on the degree of the change. A pixel for each gray level like this
, and create a histogram for each gray level as shown in FIG. 3.
したがって、活性が高く、例えば動きおよび変化の多い
受精卵では、画像演算の結果得られる画像が明るくなり
、ヒストダラムは、グレイレベルの高い方のビクセルの
数が多くなる。一方、例えばほとんど活性がなく、動き
および変化のない受精卵では、画像演算の結果得られる
画像がほぼ真黒の画像となり、ヒストグラムは、グレイ
レベルの低い方のビクセルの数か多くなるので、このヒ
ストグラム＼の配列データから、受精卵の活性度を算出
することができる。Therefore, for a fertilized egg with high activity, eg, a large amount of movement and change, the image obtained as a result of the image operation will be brighter, and the histogram will have a larger number of vixels with a higher gray level. On the other hand, for example, for a fertilized egg with almost no activity and no movement or change, the image obtained as a result of image operations will be an almost completely black image, and the histogram will have a larger number of pixels at the lower gray level. The activity level of the fertilized egg can be calculated from the sequence data of \.
なお、上述の測定中に振動笠により受精卵が動くと、正
確な測定を行うことかできない。そこで、１−記実施例
では、受精卵をマイクロマこユピュレータにより保持し
たが、例えばナイ１７ン笠のメンシュ」ユに受精卵を載
置あるいは、受精卵の上にメソシュを載置して、受精卵
を保持することもできる。Note that if the fertilized egg moves due to the vibrating shade during the above-mentioned measurement, accurate measurement cannot be performed. Therefore, in Example 1-1, the fertilized eggs were held by a micromacroupulator. It can also hold eggs.
また、上記実施例では、所定時間毎に入力される画像と
、最初に入力したマスター画像と比較したが、例えば所
定時間毎に入力される画像とその剪に入力された画像と
を比較するようにしても良い。In addition, in the above embodiment, an image input at a predetermined time interval is compared with the first input master image, but for example, an image input at a predetermined time interval and an image input at that time may be compared. You can also do it.
［発明の効果］
１−述のように、本発明の細胞の活性度測定方法では、
受精卵等の動きおよび変化の速度の遅いｉＩ！ｌ胞でも
、その活性度を正確に測定することかできる。[Effects of the Invention] 1- As mentioned above, in the cell activity measurement method of the present invention,
The movement and change speed of fertilized eggs is slow iI! It is possible to accurately measure the activity of single cells.
第１図は本発明の一実施例の活性度測定方法に用いるｙ
２置の構成図、第２図は第１図に示す装置の動作を示す
フローチャー１・、第３図はグレイレベル毎のビクセル
数を示すヒストダラムである。
１・・・・・・倒立類ｖ１鏡、２・・・・・・ＣＣＬ）
カメラ、３・・・・・・画像処理装こ。
出願人 家畜受精卵移植技術研究組合
代理人 弁理士 須 山 佐 −
第１図
グレイレベル
第３図Figure 1 shows y used in the activity measurement method of one embodiment of the present invention.
FIG. 2 is a flowchart 1 showing the operation of the apparatus shown in FIG. 1, and FIG. 3 is a histogram showing the number of pixels for each gray level. 1...Inverted class v1 mirror, 2...CCL)
Camera, 3...Image processing device. Applicant: Livestock Fertilized Egg Transplant Technology Research Association Representative Patent Attorney: Sa Suyama - Figure 1 Gray Level Figure 3
Claims (1)
に変換し、この画像信号を画像データとして記憶手段に
記憶させ、この後所定時間経過後の前記細胞の画像信号
を前記撮像手段から入力し前記記憶手段に記憶された前
記画像データと比較することを特徴とする細胞の活性度
測定方法。(1) An enlarged image of the cell is converted into an electrical image signal by the imaging means, this image signal is stored as image data in the storage means, and after a predetermined period of time has elapsed, the image signal of the cell is transferred from the imaging means. A method for measuring cell activity, which comprises comparing the inputted image data with the image data stored in the storage means.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11779887A JPS63282653A (en) | 1987-05-14 | 1987-05-14 | Method for measuring activity of cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11779887A JPS63282653A (en) | 1987-05-14 | 1987-05-14 | Method for measuring activity of cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS63282653A true JPS63282653A (en) | 1988-11-18 |
Family
ID=14720547
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP11779887A Pending JPS63282653A (en) | 1987-05-14 | 1987-05-14 | Method for measuring activity of cell |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63282653A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6429765A (en) * | 1987-07-27 | 1989-01-31 | Hitachi Ltd | Diagnosing device for number and activity of cells |
| KR100675103B1 (en) * | 2000-07-18 | 2007-01-29 | 주식회사 태산솔루젼스 | Apparatus and method for the quantitative measurement of adhesive force of cell to constructs |
| JP2012531584A (en) * | 2009-06-25 | 2012-12-10 | フェイズ ホログラフィック イメージング ペーホーイー アーベー | Egg or embryo analysis by digital holographic imaging |
| CN103583440A (en) * | 2013-11-21 | 2014-02-19 | 中国水产科学研究院黑龙江水产研究所 | Method for fast and physically distinguishing quality of mature ova of hucho taimen |
-
1987
- 1987-05-14 JP JP11779887A patent/JPS63282653A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6429765A (en) * | 1987-07-27 | 1989-01-31 | Hitachi Ltd | Diagnosing device for number and activity of cells |
| KR100675103B1 (en) * | 2000-07-18 | 2007-01-29 | 주식회사 태산솔루젼스 | Apparatus and method for the quantitative measurement of adhesive force of cell to constructs |
| JP2012531584A (en) * | 2009-06-25 | 2012-12-10 | フェイズ ホログラフィック イメージング ペーホーイー アーベー | Egg or embryo analysis by digital holographic imaging |
| CN103583440A (en) * | 2013-11-21 | 2014-02-19 | 中国水产科学研究院黑龙江水产研究所 | Method for fast and physically distinguishing quality of mature ova of hucho taimen |
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