EP1177472A2 - Videomicroscope a haute resolution destine a mesurer des echantillons extraits de suspensions de particules, avec agitation mecanique marquee des echantillons - Google Patents

Videomicroscope a haute resolution destine a mesurer des echantillons extraits de suspensions de particules, avec agitation mecanique marquee des echantillons

Info

Publication number
EP1177472A2
EP1177472A2 EP00941913A EP00941913A EP1177472A2 EP 1177472 A2 EP1177472 A2 EP 1177472A2 EP 00941913 A EP00941913 A EP 00941913A EP 00941913 A EP00941913 A EP 00941913A EP 1177472 A2 EP1177472 A2 EP 1177472A2
Authority
EP
European Patent Office
Prior art keywords
suspension
microscope
sample
particles
pulsed
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.)
Withdrawn
Application number
EP00941913A
Other languages
German (de)
English (en)
Inventor
Hajo Suhr
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Suhr Hajo Prof Dr
Original Assignee
KARL VOELKER STIFTUNG AN DER FACHHOCHSCHULE MANNHEIM
KARL VOELKER STIFTUNG AN DER F
Karl Volker Stiftung An Der Fachhochschule Mannheim
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by KARL VOELKER STIFTUNG AN DER FACHHOCHSCHULE MANNHEIM, KARL VOELKER STIFTUNG AN DER F, Karl Volker Stiftung An Der Fachhochschule Mannheim filed Critical KARL VOELKER STIFTUNG AN DER FACHHOCHSCHULE MANNHEIM
Publication of EP1177472A2 publication Critical patent/EP1177472A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/361—Optical details, e.g. image relay to the camera or image sensor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/02—Investigating particle size or size distribution
    • G01N15/0205—Investigating particle size or size distribution by optical means
    • G01N15/0227—Investigating particle size or size distribution by optical means using imaging; using holography
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/365—Control or image processing arrangements for digital or video microscopes

Definitions

  • sample suspensions of small size are an important diagnostic method.
  • suspension samples are taken from the fermentation containers, which are diluted in standardized transparent measuring chambers (for example thoma Measuring chamber) under a laboratory microscope, counted for concentration determination and measured with regard to particle size with the help of automatic image processing.
  • standardized transparent measuring chambers for example thoma Measuring chamber
  • This is a standard method in all particle-controlled processes, for example in biotechnology or in chemical process engineering, where substances in the form of microscopic particles occur in reactors and have to be characterized.
  • This invention solves two problems that have previously arisen in the microscopic diagnosis of small amounts of suspension.
  • the prepared transparent measuring chamber must be fixed and focused precisely and stably under the microscope, so that neither motion blur due to residual flow or vibrations or optical defocusing disturb the visual or automatic image evaluation. This places high demands on the microscope stand and the optical adjustment devices.
  • the sample is set in constant agitation and rapid movement in apparent paradox to the requirements of high-resolution microscopy by means of electromechanical or magnetomechanical aids. This has the effect that the sample suspension is constantly mixed again and that no concentration gradients can develop.
  • a high-resolution video microscope with extremely short exposure times of less than one microsecond is used according to the invention.
  • This video microscope is state of the art in the field of inline process control (patent DE 40 32 002 C2 and the article "In Situ Microscopy for On Line Characterization of Cell Population in Bioreactors", Biotechnology & Bioengineering 47, 106-117 (1997)) extremely short exposure time of this microscope of less than microseconds allows sharp images of moving microscopic parts none in the mechanically shaken, vibrated or stirred sample cell.
  • this microscopy concept does not require a mechanically limited precise observation volume. Instead, you can use the virtual volume, which, due to the limited depth of field of the lens image, automatically emerges from a larger suspension environment.
  • the observed measurement volume is objectively and reproducibly limited by using a numerical focus criterion in the automatic image processing algorithm. It can be calibrated with defined calibration suspensions.
  • the short-pulsed video microscope generates countable and measurable direct images from the artificially turbulent or flowing sample suspension. The counted objects can be interpreted directly as a measure of the particle concentration by calibration.
  • a larger number of images are recorded through a transparent cell wall, for example a few hundred, which are sent to the automatic image processing system for counting and measuring the particle shapes.
  • the suspension is kept in motion by miniaturized mechanical stirrers or by impressed vibrations by magneto-mechanical or electromechanical vibrations.
  • a piezo crystal that is subjected to vibrations in contact with the cuvette can be used for permanent mixing and movement of the particles.
  • the size of the statistical sample that makes up the image information is proportional to the number of images taken. However, this only applies as long as it is guaranteed that scenes of successive images (ie the random collection of particles contained therein) are completely uncorrelated.
  • the sharply depicted virtual sample volume used for evaluation within the suspension must therefore have its content between two images replace completely.
  • the mechanical shaking of the cuvette according to the invention also fulfills this purpose.
  • Undemanding sample containers A volume-precise sample chamber can be dispensed with, since the image processing only filters out the sharply imaged particles from a small part of the cuvette for evaluation anyway, thus creating a virtual but precisely calibratable sample volume.
  • Automatic image processing counts sharply imaged cells and uses the average count rate per image to generate a calibratable signal of the number concentration in the suspension. In addition, it can determine morphological details such as form factors or size histograms.
  • Simplified sample preparation The need for a strong dilution is eliminated or is alleviated, since the in situ microscope can also evaluate and calibrate images in the range of very high concentrations of, for example, over 10 9 particles per plastic. bicentimeter at 4 micron particle diameter. With existing sample quantities in milliliter format, the careful preparation of representative tiny (and therefore more error-prone) sample volumes of fewer microliters is no longer necessary, since the larger samples remain well mixed with the specimen movement and representative images are generated directly from them in the pulsed video microscope.
  • FIG. 1 shows an exemplary embodiment of the device according to the invention. It shows the microscope tube 1 with image sensor 2 (for example a CCD frame) and with objective 3 and viewing window 4 over a transparent cuboid-shaped sample cuvette 5.
  • image sensor 2 for example a CCD frame
  • objective 3 and viewing window 4 over a transparent cuboid-shaped sample cuvette 5.
  • the cuvette sits on a piezo ring 6, which is electrically excited to mechanical vibrations in a conventional manner by electrode coating can. If a suitable electrical alternating voltage 8 is applied to the electrodes, any periodic movements from low-frequency vibrations to ultrasound can be coupled into the cuvette and the mixing of the suspension can be forced.
  • a miniaturized flash lamp 7 - if necessary with condenser optics - is positioned under the cuvette in the center of the piezo ring, which generates a transmitted light exposure of the moving sample.
  • the use of pulsed luminous diodes is advantageous - because it is inexpensive.

Landscapes

  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Multimedia (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Computer Vision & Pattern Recognition (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Microscoopes, Condenser (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Analysing Materials By The Use Of Radiation (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Vidéomicroscope à haute résolution qui est utilisé pour mesurer des échantillons extraits de suspensions de particules. L'échantillon est agité et des images dudit échantillon sont prises avec un temps d'exposition extrêmement court.
EP00941913A 1999-05-13 2000-05-11 Videomicroscope a haute resolution destine a mesurer des echantillons extraits de suspensions de particules, avec agitation mecanique marquee des echantillons Withdrawn EP1177472A2 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE19923074 1999-05-13
DE1999123074 DE19923074A1 (de) 1999-05-13 1999-05-13 Hochauflösendes Videomikroskop zur Ausmessung extrahierter Proben von Partikelsuspensionen mit eingeprägter mechanischer Probenschwingung
PCT/DE2000/001466 WO2000070385A2 (fr) 1999-05-13 2000-05-11 Videomicroscope a haute resolution destine a mesurer des echantillons extraits de suspensions de particules, avec agitation mecanique marquee des echantillons

Publications (1)

Publication Number Publication Date
EP1177472A2 true EP1177472A2 (fr) 2002-02-06

Family

ID=7908570

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00941913A Withdrawn EP1177472A2 (fr) 1999-05-13 2000-05-11 Videomicroscope a haute resolution destine a mesurer des echantillons extraits de suspensions de particules, avec agitation mecanique marquee des echantillons

Country Status (4)

Country Link
EP (1) EP1177472A2 (fr)
JP (1) JP2002544560A (fr)
DE (1) DE19923074A1 (fr)
WO (1) WO2000070385A2 (fr)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101120365B (zh) * 2005-02-17 2012-01-18 辛根塔参与股份公司 种子计数和频率测量设备及方法
WO2008088249A1 (fr) * 2007-01-17 2008-07-24 Hemocue Ab Appareil de détermination de positions d'objets contenus dans un échantillon
JP6134402B1 (ja) 2016-01-29 2017-05-24 シスメックス株式会社 生体試料撮像装置及び生体試料撮像方法

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4283128A (en) * 1980-06-23 1981-08-11 The United States Of America As Represented By The Administrator Of The U.S. Environmental Protection Agency Photomicrographic system for flowing fluids
DE4032002C2 (de) * 1989-10-11 1997-05-22 Suhr Hajo Prof Dr In situ Mikroskopsonde und Meßverfahren
AT398852B (de) * 1990-02-05 1995-02-27 Kempe Eberhard Anordnung zur überprüfung des morphologischen zustands von biomassen in fermentern
WO1997004347A1 (fr) * 1995-07-19 1997-02-06 Morphometrix Technologies Inc. Balayage automatique de lamelles de microscope

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0070385A3 *

Also Published As

Publication number Publication date
DE19923074A1 (de) 2000-11-16
WO2000070385A2 (fr) 2000-11-23
WO2000070385A3 (fr) 2001-04-19
JP2002544560A (ja) 2002-12-24

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