JPS5988716A - Preparation for automatic focussing - Google Patents
Preparation for automatic focussingInfo
- Publication number
- JPS5988716A JPS5988716A JP19907982A JP19907982A JPS5988716A JP S5988716 A JPS5988716 A JP S5988716A JP 19907982 A JP19907982 A JP 19907982A JP 19907982 A JP19907982 A JP 19907982A JP S5988716 A JPS5988716 A JP S5988716A
- Authority
- JP
- Japan
- Prior art keywords
- cells
- markers
- focussing
- blood cells
- marker
- 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
Classifications
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
Landscapes
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Microscoopes, Condenser (AREA)
- Automatic Focus Adjustment (AREA)
Abstract
Description
【発明の詳細な説明】
〔発明の技術分野〕
本発明は、顕微鏡画像のパターン認識処理を行う際に、
画像入力時における自動焦点の８￥度の向上を図るため
の自動焦点用プレパラートに関するものである。[Detailed Description of the Invention] [Technical Field of the Invention] The present invention provides a method for performing pattern recognition processing on microscopic images.
This invention relates to an autofocus preparation for improving autofocus by 8 degrees when inputting an image.
自動細胞診断装置＃や自動血球分類装置など、細胞や血
球などの微小物体な姉（微鏡により光学的（二拡大し、
その光学像を掃像累子などにより電気信号に変換してそ
の画像信号を処理する装置においては、目Ｕ１焦点釉合
は必須の技術である。これらの自動装置においては、微
小物体を光学的に拡大する故に焦点深度が浅くなり、ま
たスライドガラスをｌｌＩｎ次移動して視野を変える際
に光学系の光軸と移ル１１機描の垂部ル゛が保てずに焦
点がすれ、さらにスライドガラスの厚みが場順（二より
異なるためスライドガラスの表面に塗抹された物体の光
軸方向の距離が祈、野の移動とともに変化するなどの現
象が生ずる。このため信頼度の高い画像データを得るに
は、これらの要因（二対して常（−焦点整合状態を保ち
、焦点不整合時の誤った画像データの入力を阻まなけれ
はならない。Automated cell diagnostic equipment and automatic blood cell sorting equipment are used to detect microscopic objects such as cells and blood cells (optically magnify them using a microscope).
In a device that converts the optical image into an electrical signal using a scanning device or the like and processes the image signal, focusing the eye U1 is an essential technique. In these automatic devices, the depth of focus becomes shallow because minute objects are optically magnified, and when the slide glass is moved to change the field of view, the optical axis of the optical system and the vertical part of the In addition, the distance of the object smeared on the surface of the slide glass in the optical axis direction changes with the movement of the field because the thickness of the slide glass is different from the original. Therefore, in order to obtain highly reliable image data, it is necessary to maintain a state of focus alignment and to prevent input of erroneous image data when the focus is misaligned.
ところで、自動焦点整合のアルゴリズムは、画像データ
の濃度総和の大小、画像データの空間周波数の高域成分
の大小、また隣接絵素間の差分値の自乗和の大小などに
より行なっており、これらはすべて視野内に画像が存在
するという仮定の上でｒＪ父立している。しかしながら
、スライドガラスに細胞や血球などを塗抹する際（二は
、全体Ｃ二均−（二塗抹することが困雑であり、視野に
よっては細胞や血球などが少ない場合も有り留る。した
がって、視野（＝細胞や血球などが無いか、あるいは極
めて少ない場合（二は自動焦点整合は不可能となる。By the way, the automatic focusing algorithm is based on the magnitude of the total density of the image data, the magnitude of the high-frequency component of the spatial frequency of the image data, and the magnitude of the sum of squares of the difference values between adjacent picture elements. All rJ is established on the assumption that an image exists within the field of view. However, when smearing cells, blood cells, etc. on a glass slide (2), it is difficult to smear the whole C uniformly (2), and there may be cases where there are few cells, blood cells, etc. depending on the field of view. Field of view (=If there are no or very few cells or blood cells, etc., automatic focusing is impossible.
このため、従来、上記欠点を補うため（二第１図（二示
す如く、焦点整合すべき面の近傍であるスライドがラス
１の被測定物乙の塗抹表面（二金属の蒸着などの手法（
二より規則的微小なマーク４を施し、被測定物の少ない
視野においても当該マーク４（二より焦点整合を行う方
法が採られていた。For this reason, in order to compensate for the above-mentioned drawbacks (as shown in Fig. 1), the surface of the object to be measured with the slide in the vicinity of the plane to be focused (2) is smeared on the surface of the object to be measured (2) where the slide is in the vicinity of the surface to be focused (2).
A method has been adopted in which more regular minute marks 4 are applied and focus alignment is performed using the marks 4 even in a small field of view of the object to be measured.
しかしながら、細胞や血球には厚みがありマークした口
１１が必らずしも焦点整合位＃５′ではないので、拡大
倍率が高い場合は、前述の如く焦点深度が浅いため、正
しく ＭＩＩ胞やＩＯ′１球など（−焦点を合せたこと
にはならない。また、マークは極めて薄いため（二、そ
の上（二細胞や血球などが重なるのは避けられず、画像
入力の際Ｃニマークを含めて処理することがあるため、
誤った画像データを入力してしまう恐れがあった。さら
に、金属蒸着く二よるマーキングは高価なもので、しか
も特殊な設備を要するという欠点かあった。However, cells and blood cells are thick and the marked opening 11 is not necessarily the focus position #5', so when the magnification is high, the depth of focus is shallow as described above, so it is not possible to correctly identify MII cells or IO'1 bulb, etc. (- does not mean that it is in focus. Also, since the marks are extremely thin, (2, on top of that) it is inevitable that cells, blood cells, etc. overlap, so when inputting images, do not include the C2 mark. may be processed,
There was a risk of inputting incorrect image data. Furthermore, double-sided markings made by metallization are expensive and require special equipment.
〔発明の１．」的〕
本発明は、前Ｍ［Ｈ１ｉ情（＝鋸、ｙＪてなされたもの
であり、安価でしかも、判別な設（ｉｔｆｉを必要とせ
す、かつ適正な整合を行うことのできる自動焦点用プレ
パラートを提供することを目的と−「るものである。[Invention 1. ] The present invention was developed based on the previous M[H1i situation (= saw, yJ), and is an automatic focusing device that is inexpensive, requires a discriminative installation (ITFI), and can perform proper alignment. The purpose is to provide preparations.
前記の１」的を達成するための本発明の顧１少は、被測
定物Ｃ二自動焦点用胎合マーカーを、あらかじめ混入し
たものをスライドガラス（＝ひ抹すること（二より、前
記？Ｊ５ ｆｆ１ｉ定物と自動焦点用整合マーカーとが
分１１ｉ１１ Ｌだ状態で包含されていることを偵１徴
とするものである。The first aspect of the present invention to achieve the above-mentioned objective 1 is to crush a slide glass (= grinding) a slide glass (= grinding) with a pre-mixed automatic focusing pregnancy marker. The main feature is that the J5 ff1i constant and the autofocus alignment marker are included in a state of 11i11L.
以下、本発明の一実施例を図ｍｌを参照して説明する。 Hereinafter, one embodiment of the present invention will be described with reference to Figure ml.
第２図は、本発明の日勤焦点杉−合用プレバラードの一
実力撓例を示す概略説明図である。図（−おいて、（ａ
）は平面図であり、（ｂｌは（ａｌの断面図である。１
目スライドガラスであり、２はマーカーであり、３は被
測定物（例えは卸訓ゼ１やｍ１球など）である。FIG. 2 is a schematic explanatory diagram showing an example of the strength of the day shift focus cedar-combined prevarado of the present invention. Figure (-, (a
) is a plan view, and (bl is a cross-sectional view of (al).1
The eye is a slide glass, 2 is a marker, and 3 is an object to be measured (for example, a bulletin board, an M1 ball, etc.).
以上のよう（−構成された本発明の作用（二ついて説明
する。顕微鏡などで、ａｌ胞や血球などを観察する場合
（二は、周知の如く、スライドガラスのガラス板の表面
上に細胞や血球を塗抹する工程が必要である。本発明（
二よれば、その細胞や血球を塗抹する工程の際（＝、あ
らかじめ細胞や血球に焦点整合用マーカーなる微小物体
を混入し、スライドガラスＣ′″−細胞や血球と同時に
焦点整合用マーカーを塗抹すること（二より、自動焦点
整合の際には当該マーカーによっても焦点整合を行うこ
とができる。As described above, the function of the present invention constructed as described above (- will be explained in two parts. When observing alveolar cells, blood cells, etc. with a microscope etc. (2), as is well known, cells and A step of smearing blood cells is necessary.The present invention (
According to 2, during the process of smearing the cells and blood cells (=, a microscopic object called a focus alignment marker is mixed into the cells and blood cells in advance, and the focus alignment marker is smeared on the slide glass C''' at the same time as the cells and blood cells. (Secondly, during automatic focus alignment, focus alignment can also be performed using the marker.
ここで、本発明に用いられる焦点整合用マーカー（二つ
いて述べる。本マーカーの作用は、背景との吸光夏着を
大きくして、焦点整合の検出精度を良くし、画像データ
入力後にマーカーと細胞や血球など被測定物との識別を
正しく行い、また細胞や血球などとマーカーとの重なり
を防ぎ、さらにスライドがラス塗抹の際に他のものに付
着せず（＝均一に塗抹できることを特徴としている。こ
のため焦点整合用マーカーとしては次のし、′件を、７
．ｊ、ｌｉすものならどのようなものでも良い。Here, we will discuss two markers for focus alignment used in the present invention. The function of this marker is to increase the absorption of light with the background, improve the detection accuracy of focus alignment, and connect the marker and cells after inputting image data. It is characterized by correctly identifying objects to be measured such as cells and blood cells, preventing overlap between cells and blood cells and markers, and preventing the slide from adhering to other objects during lath smearing (= uniform smearing). Therefore, the following focus alignment markers are used.
．． It can be anything as long as it is good.
■ マーカーの光のＵ及光度が被ｒｉｉｆｌ定物である
４荀胞や血球などの吸光度（二比較し、充分に大きいこ
と。■ The light intensity of the marker light must be sufficiently large compared to the absorbance of the riifl constants such as cells and blood cells.
■ 被測定物である細胞やｎｎ琢の形状に比較し大きさ
が同程度かそれ以下であり、かつ光学系・撮像系の解像
度よりも大きいこと。■ The size must be the same or smaller than the shape of the cell or nn-taku that is the object to be measured, and larger than the resolution of the optical system/imaging system.
■ 当該マーカーが細胞や血球あるいはマーカー同志で
付着しにくいこと。■ The marker is unlikely to adhere to cells, blood cells, or to other markers.
以上の要件を備える様なマーカーとして例えばラテック
ス系の乳液がある１、このようなマーカーを用いること
Ｃ二より信頼度の高い画像データを供給することができ
る。As a marker that meets the above requirements, for example, there is a latex-based emulsion (1). By using such a marker, more reliable image data can be provided.
尚、本発明は０１１記実施例（二限？されるものではな
く、本発明の要旨の勅、凹円でｆｎｌ＋々の変形例を包
含することは１うまでもない。It should be noted that the present invention is not limited to the embodiments described above, but it goes without saying that the gist of the present invention includes modifications such as fnl+ in a concave circle.
以上説明したよう（二、この発明Ｃユよると、１視野当
り数個の焦点整合用マーカーが分布するよう（二、被測
定物に混入することにより、被測定物である細胞や血球
などが無いか、少ない視野においても自動焦点整合が可
能（二なり、また焦点整合用マーカーとｎ）１胞や血球
の重なりを防ぐことができ、さら（１該マーカーがＩＩ
Ｉ Ｊ胞や血球とほぼ同じ厚さなので、４ｉ１１胞や血
球の有る視野においては、焦点整合楯度がより正確（１
行なえる自動焦点用プレパラートをＣ７（３４すること
ができる。As explained above (2. According to this invention, several markers for focus alignment are distributed per field of view (2. By mixing with the object to be measured, cells, blood cells, etc., which are the object to be measured) Automatic focusing is possible even in fields where there is no or a small field of view (2 or 2 markers for focus alignment).
Since the thickness is almost the same as that of IJ cells and blood cells, the focusing shield is more accurate (1
C7 (34 autofocus preparations) can be used.
第１図は、従来行なわれていたマーキングな流点整合用
マーカーが塗抹された状態の火施例であり、
へ、第２図（ａ）は正面図、（ｂ）は断面図である。
１・・・スライドガラス、 ２・・・焦点整合用マーカ
ー、 ３・・・被測定物、 ４・・・マーク。
代理人 弁理士 則 近 憲 佑（ほか１名）（７）
第 １ 図
（ｂ）
〜
第２図
（０）
□−−−−□−ｊ４２苓λ−１
９３FIG. 1 shows an example of the flow point alignment marker with conventional markings smeared on it, and FIG. 2(a) is a front view, and FIG. 2(b) is a sectional view. DESCRIPTION OF SYMBOLS 1... Slide glass, 2... Focus alignment marker, 3... Measured object, 4... Mark. Agent Patent attorney Noriyuki Chika (and 1 other person) (7) Figure 1 (b) - Figure 2 (0) □----□-j42蓓λ-1 93
Claims (1)
たものをスライドガラスに塗抹すること（二より、前記
被６１１ｊ定物と自動焦点用整合マーカーとが分離した
状態で包含されていることを特徴とする自動角店用プレ
パラート。Smearing a slide glass with an object to be measured mixed with an automatic focus alignment marker in advance (Secondly, the object to be measured and the automatic focus alignment marker are included in a separated state) Automatic corner store preparation parts.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19907982A JPS5988716A (en) | 1982-11-15 | 1982-11-15 | Preparation for automatic focussing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19907982A JPS5988716A (en) | 1982-11-15 | 1982-11-15 | Preparation for automatic focussing |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS5988716A true JPS5988716A (en) | 1984-05-22 |
Family
ID=16401755
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP19907982A Pending JPS5988716A (en) | 1982-11-15 | 1982-11-15 | Preparation for automatic focussing |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5988716A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1987002803A1 (en) * | 1985-11-04 | 1987-05-07 | Cell Analysis Systems, Inc. | An apparatus and method for analyses of biological specimens |
| US4998284A (en) * | 1987-11-17 | 1991-03-05 | Cell Analysis Systems, Inc. | Dual color camera microscope and methodology for cell staining and analysis |
| US5016283A (en) * | 1985-11-04 | 1991-05-14 | Cell Analysis Systems, Inc. | Methods and apparatus for immunoploidy analysis |
| US5086476A (en) * | 1985-11-04 | 1992-02-04 | Cell Analysis Systems, Inc. | Method and apparatus for determining a proliferation index of a cell sample |
| US5109429A (en) * | 1985-11-04 | 1992-04-28 | Cell Analysis Systems,Inc. | Apparatus and method for analyses of biological specimens |
| US5134662A (en) * | 1985-11-04 | 1992-07-28 | Cell Analysis Systems, Inc. | Dual color camera microscope and methodology for cell staining and analysis |
| DE10250247A1 (en) * | 2002-10-28 | 2004-05-13 | Leica Microsystems Heidelberg Gmbh | Microscope slide and method for making a slide |
| JP2008209258A (en) * | 2007-02-27 | 2008-09-11 | Honda Electronic Co Ltd | Sample support for acoustic parameter measuring device, and acoustic parameter measuring device |
| WO2012025220A1 (en) * | 2010-08-23 | 2012-03-01 | Euroimmun Medizinische Labordiagnostika Ag | Method and device for focusing substrates in an automated manner in fluorescence microscopy |
| JP2013254108A (en) * | 2012-06-07 | 2013-12-19 | Canon Inc | Defocus amount estimating method, imaging device, and translucent member |
-
1982
- 1982-11-15 JP JP19907982A patent/JPS5988716A/en active Pending
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5134662A (en) * | 1985-11-04 | 1992-07-28 | Cell Analysis Systems, Inc. | Dual color camera microscope and methodology for cell staining and analysis |
| US4741043A (en) * | 1985-11-04 | 1988-04-26 | Cell Analysis Systems, Inc. | Method of and an apparatus for image analyses of biological specimens |
| JPS63501597A (en) * | 1985-11-04 | 1988-06-16 | セル・アナラシス・システムズ・インコ−ポレ−テッド | Analytical methods and devices for biological specimens |
| WO1987002803A1 (en) * | 1985-11-04 | 1987-05-07 | Cell Analysis Systems, Inc. | An apparatus and method for analyses of biological specimens |
| US5016283A (en) * | 1985-11-04 | 1991-05-14 | Cell Analysis Systems, Inc. | Methods and apparatus for immunoploidy analysis |
| US5086476A (en) * | 1985-11-04 | 1992-02-04 | Cell Analysis Systems, Inc. | Method and apparatus for determining a proliferation index of a cell sample |
| US5109429A (en) * | 1985-11-04 | 1992-04-28 | Cell Analysis Systems,Inc. | Apparatus and method for analyses of biological specimens |
| US4998284A (en) * | 1987-11-17 | 1991-03-05 | Cell Analysis Systems, Inc. | Dual color camera microscope and methodology for cell staining and analysis |
| DE10250247A1 (en) * | 2002-10-28 | 2004-05-13 | Leica Microsystems Heidelberg Gmbh | Microscope slide and method for making a slide |
| DE10250247B4 (en) * | 2002-10-28 | 2006-06-01 | Leica Microsystems Cms Gmbh | Sample carrier for microscopy and method for preparing a sample carrier |
| US7583436B2 (en) | 2002-10-28 | 2009-09-01 | Leica Microsystems Cms Gmbh | Sampler carrier for a confocal microscope and method for fabricating a sample carrier |
| JP2008209258A (en) * | 2007-02-27 | 2008-09-11 | Honda Electronic Co Ltd | Sample support for acoustic parameter measuring device, and acoustic parameter measuring device |
| WO2012025220A1 (en) * | 2010-08-23 | 2012-03-01 | Euroimmun Medizinische Labordiagnostika Ag | Method and device for focusing substrates in an automated manner in fluorescence microscopy |
| CN103080807A (en) * | 2010-08-23 | 2013-05-01 | 欧蒙医学诊断技术有限公司 | Method and device for focusing substrates in an automated manner in fluorescence microscopy |
| JP2013254108A (en) * | 2012-06-07 | 2013-12-19 | Canon Inc | Defocus amount estimating method, imaging device, and translucent member |
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