DE102005018845A1 - Miniature multifunctional opto sensor control and lock unit for minimal invasive sensor location has measurement chamber and micrometer drive - Google Patents

Abstract

A miniature multifunctional opto sensor control and lock unit for minimal invasive sensor location provides continuous physical, chemical and biological analysis of fluids and gases flowing through a closed or open measurement chamber (16) with sensor mounting holes (1-9) and a micrometer drive (14).

Description

A miniaturized and multifunctional optosensory measuring and control sluice for minimally invasive process analysis with high spatial usnd temporal resolution in biological and abiotic systems and for experimental Manipulation physico-chemical and biological parameters represents an innovative tool that over conventional measuring devices provides key benefits for the purpose of in situ bio-process analysis.

Progress in quantifying life processes under natural environmental conditions in the field as well as under controlled conditions in the laboratory today requires the use of techniques and sensors that are characterized by miniaturization, precision, longevity and minimal disruption of the system under investigation. The contact-free and therefore also contamination-free measurement is the goal of the minimally invasive examination, for which optical sensors are capable for the first time. Miniaturization and minimally invasive intervention in the system under investigation are two essential prerequisites for the progress of the understanding of the coordinated course of life processes in spatially and temporally high resolution. Fiber-optic sensors with a diameter of only a few micrometers (optodes) represent the current state of the art of measuring on a microscale scale for the quantification of biologically relevant physico-chemical parameters, such as oxygen partial pressure (PreSens Precision Sensing GmbH, Regensburg). Furthermore, the temperature, the CO ₂ partial pressure, the pH or the chlorophyll content of green plants can be optically quantified.

Hybridoptodes represent the latest generation of optical sensors whose market launch is expected within one to two years (HYBOP research collaboration project of the BMBF Priority Program BIOPHOTONIK at the University of Düsseldorf, coordinated by the applicant). Hybrid-optic devices can use analyte-specific microparticles to measure two physicochemical parameters at the same time and on a molecular scale. The hybrid character halves the number of sensors needed for a process analysis, thereby providing significant methodological progress. For example, with only one CO ₂ -O ₂ hybridoptode, the respiratory quotient of cell cultures, tissues and organs can be determined from the simultaneous measurement of metabolic O ₂ consumption and CO ₂ production.

in the Unlike conventional sensors, the new optical one stands out Sensor technology in addition to the miniaturization mainly by the independence the measurement of the state of aggregation of the analyte. With optical Sensors are measurements in the aqueous and gaseous Phase alike possible, whereas conventional measuring methods be defined by the state of aggregation of the analyte. this will in the following for biologically relevant parameters explained.

In the aqueous phase, CO ₂ and O _{2 are} measured electrochemically by means of electrodes (eg Clark-type O ₂ electrode or CO ₂ electrode from Microelectrodes Inc., Bedford, USA). The disadvantage of electrochemical measurements lies in the consumption of a certain amount of analyte by the measuring process itself, so that an unaffected by the measuring instrument process analysis is not guaranteed. Likewise, the long-term stability of electrochemical sensors is limited.

Measurements in the gas phase are carried out with gas analyzers (absolute or differential measurements) or gas chromatographs suitable for trace gas analysis. Conventionally, CO ₂ analysis uses infrared gas analyzers (IRGA) with accuracies in the range of ± 1-2 ppm. For the accuracy of measurement by means of gas analyzers, however, defined mass flow rates are crucial, which are produced with a complex measuring and control technology. Therefore, such systems are very expensive and maintenance intensive. Spatial high-resolution measurements in tissues and organs can not be carried out either. Since they are open measuring systems, they can not be used under sterile conditions. The gas analysis with gas chromatographs is technically complex, costly and time-consuming. Furthermore, their use for field measurements can be accomplished only with high infrastructural effort (laboratory trolley, power supply, explosion protection, etc.).

Pulsed amplitude modulation (PAM) fluorometer (Walz, Effeltrich, Germany, LI-COR Inc., Lincoln, USA) for the physiological characterization of the photosynthesis of green plants corresponds to the state of the art in the field of bio-process analysis. In plant science, chlorophyll fluorescence analysis has become established as a powerful method. Restrictive, however, the actual photosynthetic O ₂ production and the CO ₂ fixation is not measured in this method, but calculated from the fluorescence intensity as a measure of the photochemical charge separation rate of the antenna pigments of the photosystems. A hybrid optode for the simultaneous measurement of chrolophyll content and O ₂ production is a viable quantitative and at the same time much cheaper alternative to the fluorometric method ren.

electronic Temperature measurements are usually made with absolute temperature sensors (Pt 100, Pt 1000) or reference-dependent thermocouples. With the latter will be a miniaturization up to about 0.2 mm in diameter (e.g., World Precision Instruments Inc., Sarasota, USA), wherein however, an additional one Referenzmeßstelle the Absolute temperature is required. In contrast, with temperature-sensitive Microparticles spatially high-resolution measurements the temperature is carried out be only on the dimensions of the wearer of the temperature-sensitive Coating depends.

• 1) multipler Mikrosensorik,
• 2) minimalinvasiver Präparation und Sensorapplikation zur online Messung unter in situ Bedingungen und
• 3) experimenteller Manipulation im Freiland und Labor, wie sie in der vorliegenden Meß- und Regelschleuse realisiert ist, liegt nach dem Ergebnis der fachlichen Recherche des Antragstellers bisher nicht vor.

The measuring principle underlying the sensors decisively defines the technical design of closed measuring cells or flow measuring cells with multiple sensor components. By way of example, mention may be made of the embodiment of a multiple sensor device (Publication Pub. No .: US 2005/0016260 A1, Jan. 27, 2005), which is used for the physicochemical analysis of liquids, but is unsuitable for the analysis of gases. Corresponding to this example, there are a number of publications that present different solutions of multiple sensor measurement. However, these devices are usually designed as monophasic systems, either for gaseous or aqueous material analysis. Furthermore, multiple sensor systems are designed for the purpose of analysis only as closed measuring cells or as flow measuring cells. An integrative technical solution that, beyond the function of a measuring cell, creates a functional synergy

• 1) multiple microsensors,
• 2) minimally invasive preparation and sensor application for online measurement under in situ conditions and
• 3) experimental manipulation in the field and laboratory, as it is realized in the present measuring and control sluice, according to the result of the expert search of the applicant so far not available.

• 1) die durch den Aggregatszustand des Analyten festgelegten Meßprinzipien und den daraus resultierenden Unzulänglichkeiten der Messung sowie den stark eingeschränkten Einsatzmöglichkeiten;
• 2) die unzureichende Miniaturisierung für den minimalinvasiven Einsatz.

In summary, two specific technical deficiencies, which do not meet the requirements of modern bio-process analysis, result from the mentioned specific deficiencies of conventional sensors and associated multiple measuring cells:

• 1) the measuring principles defined by the state of aggregation of the analyte and the resulting inadequacies of the measurement as well as the severely restricted possibilities of use;
• 2) insufficient miniaturization for minimally invasive use.

As an example, the measurement of the CO ₂ partial pressure in the leaf tissue of higher plants may be mentioned. Although CO _{2 is} the substrate of photosynthesis of all green plants, so far there is no possibility of direct measurement of the CO ₂ partial pressure in the leaf interior. It is calculated approximately from the atmospheric CO ₂ partial pressure and the stomatal conductivity. Optical sensors, which are introduced by means of a lock into the gas space of the leaves, provide an adequate solution to this problem. Considering the nature of CO ₂ as a greenhouse gas with climate relevance, progress in understanding the spatial and temporal heterogeneity of photosynthetic CO ₂ fixation as a bioprocess in green plants under natural conditions is of great importance.

Technical problem

• a) ihrer sicheren Applikation sowie der mikroskalig exakten und reproduzierbaren Positionierung in dem zu untersuchenden Objekt;
• b) dem Schutz vor Beschädigung der Sensoren durch mechanische Beanspruchung während des Meßvorgangs, eine entscheidende Voraussetzung für Langzeituntersuchungen (Wochen – Monate);
• c) der Erfordernis zur Applikation mehrerer Sensoren unterschiedlicher Analytspezifität auf kleinstem Raum;
• d) der Messung mehrerer Parameter in einem sehr kleinen Probenvolumen (nur wenige Mikroliter);
• e) der mikromanipulativen Präparation unter sensorischer Kontrolle und Wahrung der Sterilität.

Microsensors, in particular optical microsensors (optodes) are due to their extreme miniaturization very sensitive measuring instruments, which may not be exposed to mechanical stress during application and during the measuring process. This results in numerous problems for the user when carrying out minimally invasive process analyzes with respect to:

• a) their secure application as well as the micro-scale exact and reproducible positioning in the object to be examined;
• b) the protection against damage of the sensors due to mechanical stress during the measuring process, a crucial condition for long - term investigations (weeks - months);
• c) the requirement for the application of multiple sensors of different analyte specificity in the smallest space;
• d) the measurement of several parameters in a very small sample volume (only a few microliters);
• e) the micromanipulative preparation under sensory control and maintaining sterility.

Want the user over Beyond online measurements, perform experimental manipulations, including sampling and / or cloth application, so must always a special device be created, which meets the respective metrological requirements Takes into account. This is especially true for Outdoor investigations under variable environmental conditions.

The Solving specific methodological problems is technically demanding Time-consuming and cost-intensive and can be in many places lack of infrastructure (fine mechanical and electrical workshops) and professional qualification can not be realized.

The multifunctional conception of the optosensory measuring and control lock (Omni-Lock) provides an integrative technical solution that meets those requirements Fulfills and thus the user a broad range of methods to minimally invasive process analysis opened.

Technical Troubleshooting

One massive cylindrical body (Sensor body) with horizontal circumferential groove in the lower half and bevelled upper edge (Inclined surface, 45 ° inclined) becomes rotatable and sealing in a cylindrical tray (carrier plate) snugly used. In the lower planar surface of the sensor housing is Centrally a small cavity in the form of a hemisphere (measuring chamber) sunk. Located at a small concentric distance to the circumference of the measuring chamber a puncture with sealing ring. One in the bottom surface of the sensor housing Completely sunken annular Peltier element, whose inner opening adjacent to the sealing ring, allows the electronic regulation the temperature in the measuring chamber and in the system under investigation in the area of the contact surface of the Measuring and Usually lock. The Peltier element is controlled by a PID controller (proportional, integral and differential), the three modes of operation of the Temperature control allows: 1) a synchronous with the ambient temperature Caster regulation, 2) one of the ambient temperature by one constant amount deviating follow-up regulation and 3) one of the ambient temperature independent constant temperature specification. These options for thermal control are mainly for the process analysis in the field of great Importance.

On the circular wall of the carrier plate are three offset by 120 ° crescent Slats mounted, which pivot horizontally under a guide ring are stored. The rotatability of the sensor housing in the carrier plate is achieved by in which the slats are pivoted into the groove of the sensor housing. This must be the sensor housing be pressed with light pressure on the bottom plate of the carrier plate, whereby the measuring chamber is sealed to the bottom plate. The guide ring serves as Abutment.

Remains the bottom of the carrier plate closed, so is the Omni-Lock as a closed cell or as a flow cell too operate (see below). Is the measuring chamber via a closable opening in the bottom plate with a biological or abiotic system in Connection, then the Omni-Lock acts as a control lock. About the central opening in the bottom plate are sensors and micro tools through the measuring chamber positioned in the examination subject. To guarantee a sealing connection between lock and examination object First, the carrier plate with the help of a specially designed eccentric clamping device fixed on the object. Thereafter, the sensor housing is in the carrier plate used.

The sensor housing is provided with several holes of different diameters. A central Bore in the vertical axis of the sensor housing meets the measuring chamber at the apex of the calotte. Further holes are radial symmetric in the Angle of 45 ° to Vertical arranged and lead at equal intervals from the inclined surface of the sensor housing the calotte of the measuring chamber. With one exception, the axes of all holes have the geometric Center of the measuring chamber as a common point of intersection. The exception is a hole whose Axis cuts the 'equator' of the measuring chamber. Provided with a valve, this hole acts as an outlet to the residue-free filling of the measuring chamber with liquids or gases in vertical position. The removal of gas or liquid samples as well as the application of chemical substances also takes place via this opening. Several of the radial symmetry arranged bores act as cannulas, through the sensors and / or Micro tools from outside into the measuring chamber and introduced into the examination subject and positioned gas-tight become. By means of a micrometer feed all sensors and micro tools can be used individually along the drilling axes moved exactly and reproducibly back and forth become. Two additional holes are also equipped with valves. One is for filling the measuring chamber with liquids and gases while the other for the Pressure regulation in the measuring chamber to disposal stands.

• a) als geschlossene Meßzelle zur simultanen Messung physiko-chemischer und biologischer Parameter in einem sich in der Meßkammer befindlichen gasförmigen oder flüssigen Medium. In diesem Modus ist die Öffnung des Trägertellers verschlossen und der Stoffaustausch in der Meßzelle erfolgt über die Ein- und Auslaßventile. Die Sensoren werden auf der Kalotte der Meßkammer positioniert. Da nur der Eigendurchmesser der Sensoren den minimalen Abstand zueinander auf der Kalotte definiert, kann durch den Einsatz von Hybridoptoden die Zahl der in derselben Probe gleichzeitig zu erfassenden Parameter auf engstem Raum verdoppelt werden. Diese Meßanordnung eignet sich besonders zur Analyse sehr kleiner, nur wenige Mikroliter betragende Probenmengen eines gasförmigen oder flüssigen Stoffes oder Stoffgemisches, das in die Meßkammer eingeleitet wird.
• b) als Durchflußmeßzelle mit geschlossenem Trägerteller und geöffneten Ein- und Auslaßventilen oder im Bypass-Modus. Hierzu ist die Meßkammer bei geschlossenen Ventilen aber geöffnetem Trägerteller mit dem Untersuchungsobjekt verbunden und der Stofftransport erfolgt als Bypass vom Untersuchungsobjekt durch die Meßkammer und wieder zurück.
• c) als Regelschleuse zur ortsidentischen, sukzedanen Messung physiko-chemischer und biologischer Parameter und Mikropräparation in biologischen oder abiotischen Systemen. In dieser Konfiguration ist die Omni-Lock über die Öffnung im Trägerteller mit dem Untersuchungsobjekt verbunden. Mittels Mikrometervorschub werden die auf der Schrägfläche des Sensorgehäuses angeordneten Sensoren und Mikrowerkzeuge nacheinander in das Untersuchungsobjekt eingeführt. Die Drehbarkeit des Sensorgehäuses ist die konstruktive Voraussetzung zur freien Wahl der Meßstelle im Untersuchungsobjekt auf einem Umkreis, dessen Länge sich aus dem Winkel der Kanülen zur Horizontalen und der Tiefe der Meßstelle im Objekt ergibt. Dadurch kann mit zunehmender Tiefe ein dreidimensionales Raster von Meßpunkten erstellt werden, das sich kegelförmig in das Untersuchungsobjekt hinein erstreckt. Über die vertikale Kanüle des Sensorgehäuses kann ein weiterer Sensor senkrecht zur Oberfläche in das Untersuchungsobjekt appliziert werden, wodurch Messungen entlang der Rotationsachse des kegelförmigen Meßsektors möglich sind. Dies ist die einfachste Variante zur Erfassung einer Meßgröße in einem Untersuchungsobjekt, entweder in konstanter Tiefe oder im vertikalen Gradienten. Darüber hinaus dient die vertikale Kanüle der exakten Positionierung von Mikroendoskopen und -werkzeugen für die Mikropräparation im Untersuchungsobjekt.

By choosing the center of the measuring chamber as a common point of intersection of the cannulas, the measuring and control sluice offers three different uses for process analysis:

• a) as a closed measuring cell for the simultaneous measurement of physico-chemical and biological parameters in a gaseous or liquid medium located in the measuring chamber. In this mode, the opening of the carrier plate is closed and the mass transfer in the measuring cell via the inlet and outlet valves. The sensors are positioned on the calotte of the measuring chamber. Since only the intrinsic diameter of the sensors defines the minimum distance to each other on the dome, the use of hybrid optics can double the number of parameters to be recorded simultaneously in the same sample in the smallest possible space. This measuring arrangement is particularly suitable for the analysis of very small, only a few microliters amount of sample amounts of a gaseous or liquid substance or mixture of substances in the measuring chamber is directed.
• b) as a flow cell with closed carrier plate and open inlet and outlet valves or in bypass mode. For this purpose, the measuring chamber is connected with closed valves but open carrier plate with the object to be examined and the mass transfer takes place as a bypass from the examination subject through the measuring chamber and back again.
• c) as a control lock for the location-specific, successive measurement of physicochemical and biological parameters and micro-preparation in biological or abiotic systems. In this configuration, the Omni-Lock is connected to the object to be examined via the opening in the carrier plate. By means of micrometer advance, the sensors and micro-tools arranged on the inclined surface of the sensor housing are successively introduced into the examination subject. The rotatability of the sensor housing is the design prerequisite for the free choice of the measuring point in the examination object on a circumference, the length of which results from the angle of the cannulas to the horizontal and the depth of the measuring point in the object. As a result, with increasing depth, a three-dimensional grid of measuring points can be created, which extends conically into the examination subject. Via the vertical cannula of the sensor housing, a further sensor can be applied perpendicular to the surface in the examination subject, whereby measurements along the axis of rotation of the conical measuring sector are possible. This is the simplest variant for detecting a measured quantity in an examination object, either at a constant depth or in a vertical gradient. In addition, the vertical cannula serves the exact positioning of microendoscopes and tools for the micro-preparation in the examination subject.

In the following, a specific technical design of the optosensory measuring and control sluice is described and in its mode of operation using the example of the application of optical microsensors (O ₂ - and CO ₂ optodes) in the sapwood of trees to quantify the gas partial pressures while avoiding embolisms and thus the maintenance of the transpiration current.

The overview drawing ( 1 ) illustrates in a simplified representation of the essential components of the measuring and control lock. In the cross-sectional profile (upper diagram), the rotatable sensor housing with measuring chamber and Peltier element can be seen, which is inserted into the support plate and locked with the 3-part lamellar shutter. A gaseous or liquid mass transfer between the measuring chamber and a living or abiotic system (lock function) takes place via a small opening in the base plate. Microsensors are introduced gas-tight through cannulas in the sensor housing into the measuring chamber. With the aid of a convertible micrometer feed, all microsensors and tools can be precisely positioned through the measuring chamber in the examination subject. The arrangement of the sensors in the sensor housing allows the creation of a conical grid of measuring points, which extends with increasing diameter in the depth of the examination subject. The top view (lower diagram) shows the clock-shaped housing of the carrier plate with the crescent-shaped lamellae and an optional assignment of the measuring chamber with sensors. The inlet and outlet valves and the opening for pressure regulation in the measuring chamber are also shown.

The sensor housing (d = 30 mm), here made of PEEK plastic, is equipped with a total of nine holes of different diameters (d = 1 - 2.4 mm). The central bore (d = 2.4 mm) in the vertical axis of the sensor housing meets the measuring chamber (d = 5 mm, V = 35) at the vertex of the calotte. Another eight holes are arranged radially symmetrically at an angle of 45 ° to the vertical and lead at equal intervals from the inclined surface of the sensor housing on the calotte of the measuring chamber ( 2 ). Seven of these holes have the geometric center of the measuring chamber as a common point of intersection. Only hole no. 9 hits the equator of the measuring chamber ( 2A ). It is used for the discharge of liquids and gases and at the same time the sampling and substance application. For this dual function, a sealable infusion cannula with lateral septum is used. Five of the radial symmetry holes (Nos. 1-5) act as cannulas through which sensors (optodes and hybrid optics, thermocouples and electrodes) and micro tools are introduced from the outside into the measuring chamber and into the examination subject and gas-tight. Bore No. 7 carries a valve and is used to fill the measuring chamber with liquids and gases, while Bore No. 8 is provided with an adjustable pressure relief valve for pressure regulation in the measuring chamber.

The carrier plate (d = 40 mm) made of anodized aluminum has the shape of a watch case. Bottom plate (thickness = 1 mm), wall (height = 5 mm, thickness = 5 mm) and lateral supports for the web for fastening the tensioning belt and a groove for receiving a suspension clip of the tension belt are made of one piece ( 3 ). For use of the Omni-Lock as a measuring cell, the bottom plate of the carrier plate remains closed; For use as a lock, holes up to 5 mm in diameter can be placed depending on the intended use. The three hinged blades are hori zontal rotatably mounted on the wall of the carrier plate. They are dimensioned so that they encompass almost the entire circumference of the groove of the sensor housing. This is the prerequisite for the rotation of the sensor housing while ensuring the sealing of the measuring chamber to the bottom plate. A guide ring over the slats serves as an abutment for absorbing the contact pressure of the sensor housing in the carrier plate.

The mounting of the carrier plate on the object - here a tree trunk - takes place by means of an eccentric tensioner as a special embodiment ( 4 ). In a 6 mm thick aluminum plate (clamping plate: height = 25 mm, width = 41 mm) are milled two rectangular openings at the same distance from the vertical axis, in each of which an eccentrically mounted knurling roller is inserted. The knurling roller is mounted on an axle, which is rotatably mounted in the clamping plate. A spiral spring rotates the axis and thereby causes a pressure of the knurling roller on the inner opening surface. To perform a tension belt between the inner surface of an opening and the knurled roller it is pivoted with a knurled handle against the Federandruck to the outside. Due to the permanent pressure of the eccentric knurling roller on the tension belt, it can only be pulled in one direction, but the back-up always remains blocked. The clamping plate is guided by two studs, which are firmly mounted on an abutment. A central knurled screw moves the clamping plate back and forth over the stud for a distance of 8 mm. This device allows a quick mounting of the carrier plate on the examination object by two polyester bands on both sides enclose the object and are biased by the eccentric rollers. By the displacement of the clamping plate on the stud bolt a uniform tensile force is exerted on both bands in a short way, whereby the carrier plate is locked without displacement on the examination object. In addition, this embodiment of the clamping device has the advantage that it is suitable for mounting the Omni-Lock on very differently shaped objects of any size, even if the diameter is significantly smaller than that of the carrier plate.

There Microsensors and especially optodes exposed to no mechanical forces be allowed to it is impossible such sensors unprotected in animal or plant tissues and organs, which often prevents the user from using it at all. A solution The Omni-Lock offers this problem in such a way that with this Device fine cannulas introduced into tissues and organs can be in which then the sensors are pushed up to the cannula tip. About the cannulas the measuring chamber is the Omni-Lock connected to the examination object.

In Plants is the application of optical microsensors by solid Graduation-, Hardening and supporting tissue, but above all made difficult by the woodiness. Furthermore is for minimally invasive access to the juice-bearing plant tissues the avoidance of emboli by the ingress of air during the Sensor application a crucial methodological requirement to the tissue function during to maintain the in situ measurement. Vessels in which to apply of sensors without air exclusion of under negative pressure water thread breaks through embolism, stand for further Investigations no longer available. The Omni-Lock solves this methodological problem by the possibility for the application of cannulas and sensors under air exclusion in the liquid Phase. For this purpose will be during of introduction a cannula In the tissue to be examined, degas the measuring chamber with one Liquid, e.g. a sterile, aqueous isotonic Solution, flows through. For this becomes a continuous fluid circuit between the inlet and outlet of the measuring chamber produced. By applying a controllable overpressure in the measuring chamber is the penetration of air into the tissue during the sensor application and the measurement is prevented. This method of access in woody Vegetable tissues under exclusion of air for the purpose of embolism prevention has already been successfully tested by the applicant [Gansert D, Burgdorf M, Lösch R (2001) A novel approach to the in situ measurement of oxygen concentrations in the sapwood of woody plants. Plant, Cell and Environment 24: 1055-1064]. It became the purpose of multifunctionality in the technical realization the Omni-Lock considered.

Microendoscopes and tools are used for the micro-preparative procedure, eg for the removal of tissue samples or the application of probes. These, like the sensors, are positioned in the object by means of a micrometer feed. In the present functional model a built-in measuring screw (adjustment range: 0-25 mm, accuracy: 0.002 mm) with stationary spindle is used ( 5 ). On the spindle, a clamping plate for fixing of Senso ren and micro tools is placed. The clamping plate is guided on two rods (d = 3 mm, 1 = 41 mm), which are firmly connected to the transducer plate for the micrometer and a base plate. The base plate also serves to lock the feed on a sensor socket with a sealing ring in the sensor housing, through which a sensor is inserted into the measuring chamber and into the examination object. Sensor socket, base plate and clamping plate are centered on one axis. After positioning, the sensor is released from the clamping plate and festge the sensor socket by turning the feed pulled. As a result, the sensor is locked gas-tight in the housing. Thereafter, the locking of the base plate is released at the sensor socket and removed the micrometer feed. One after the other, all sensors are positioned and sealed. This technical implementation of the micrometer feed has three major advantages: 1) weight savings in the metering and control gate, 2) the sensor cables are not twisted during positioning as they are moved without self-rotation, 3) cost savings by producing only a standardized micrometre feed for all Connecting sockets on the sensor housing is made to fit.

1

1-9

drilling with different diameters for:

 1

temperature sensor

 2

pH O ₂ hybridoptode

3

pH microelectrode

4

TO ₂ hybridoptode

5

O ₂ optode

6

CO ₂ -O ₂ hybridoptode and micro tools

7

liquid and gas inlet

8th

pressure regulation

9

liquid and gas outlet, sampling and substance application

10

swiveling Slat of the 3-part lamellar shutter in the carrier plate

11

Annular Peltier element

12

rotatable sensor housing

13

microsensors and tools

14

Reversible Micrometre feed for Microsensors and tools

15

microsensors and tools

16

measuring chamber

17

microsensors and tools

18

O-ring to seal the measuring chamber

19

carrier plate with three-piece louvre closure

20

living or abiotic system

2

A

Sensor housing: cut with holes 3, 6, 9

16

measuring camera

B

cut with holes 2, 4, 6

C

At sight with positions of holes 1-9

3

A

Backing pad: At sight

10

swiveling lamella

22

web for tension belt

B

sideview

21

Closable opening in the bottom plate of the carrier plate

23

groove for suspension bracket

C

Hook-in hanger for tension belt

4

A

Eccentric: front view

24

eccentric mounted knurling roller

25

knurled

B

At sight

26

thumbscrew

27

chipboard

28

abutment with notched bearing surface

5

A

Micrometer feed: sideview

29

Micrometer head with stationary (not rotating) spindle

30

guide rod for the clamp

B

pickup the built-in micrometer

C

clamp for locking microsensors and tools

31

locking screw

D

baseplate for fixing the micrometer feed on hexagonal

Sensor socket head

Claims (3)

The claim is levied for a miniaturized and multifunctional optosensory measuring and control lock for the minimally invasive application of microsensors and micro tools in living and abiotic systems for quantitative, spatially and temporally high-resolution process analysis, preparation and experimental manipulation under field and laboratory conditions. In the function of a lock, the technical prerequisite for the continuous physico-chemical and biological analysis of gaseous or liquid substances in the flowing or stationary state while maintaining communication with the respective biological or abiotic system is fulfilled. In addition to the simultaneous and location-identical measurement of several biologically relevant parameters (eg T, pH, O ₂ , CO ₂ , chlorophyll), the use of optical microsensors allows for the first time a process analysis independent of the state of aggregation. Through experimental manipulation and minimally invasive intervention on the examination subject under sensory control, without interruption of the measurement procedure and under sterile conditions, the measuring and Regelschleuse an integrative technical solution for the causal analysis of biological and physico-chemical processes, as it was not previously available. According to claim 1, several microsensors are different analyte specificity, in particular optical sensors (optodes and hybrid optics), in one housed around a shaft rotatable, gas-tight sensor housing in radial symmetry arrangement and by micrometre feed individually in a miniaturized measuring chamber positioned. The chamber optionally works as a) closed measuring cell, b) as a flow cell and c) as a controllable lock, through which sensors and micro-tools, e.g. Micro scissors and micro endoscopes, in a biological or abiotic System can be positioned in variable depth. By adapting the physicochemical (Gaseous or liquid Phase, temperature, pressure, pH etc.) and biological conditions (isotonicity, nutrients, sterility etc.) in the lock to the respective system to be examined and the Use of optosensory micro-metering is a minimally invasive in situ process analysis guaranteed. moreover is the measuring and rule lock autoclavable, so that it meets microbiological sterility claims. The technical design of the measuring and control lock complied with claim 1 the requirements for a wide range of applications for process analysis in the field and in the field Laboratory by means of: 1) multifunctional use by: a) Miniaturization (chamber volume: 35 μl), b) Function as closed measuring cell, Flow cell and adjustable lock in contact with the examination object, c) easy and quick mounting on the examination object (plug & play) by two-piece System structure consisting of a rotatable sensor housing and carrier plate with lamellar closure, d) Use in the field and laboratory, under atmospheric and aquatic conditions; 2) simultaneous measurement of several physico-chemical and biological Parameters in the same sample: a) biotic or abiotic nature, b) in direct communication with the examination object (Bypass mode), b) in gaseous form or liquid Status, c) in flowing or dormant phase; 3) minimally invasive sensor application and micro-preparation under sensory control and sterile conditions; 4) individual Positioning of sensors and tools by micrometer feed without cable twisting; 5) Ensuring long-term series of measurements through secure sensor housing and maximum protection of the sensors before mechanical stress; 6) experimental manipulation at the examination object by change Physico-chemical and biological parameters under online measurement; 7) pressure-compensated sampling and substance addition.