Shear Force Generating Device for Analysis of Biopsied Mammalian Samples

Abstract

A shear force generating device includes a shear-force generator structured for exerting a shear force at to each volume element of a sample of body fluid or tissue and a native conformation prion protein arranged in a container. A controller controls the shear-force generator to exert a shear-force of at maximum 10% of one shear-force intensity to each volume element of the sample arranged in the container. A light source is directed into the container. An optical detector is arranged to detect light emanating from the container. The detector is coupled to a computer including code to analyze light detected by the optical detector as a measure for the content of aggregated prion protein by comparing the light detected to a Reference Information Database.

Claims

1. Device for analysis of a biopsied mammalian sample of body fluid or tissue and a native conformation prion protein to determine the presence of disease-related aggregated conformation prion protein, the device comprising a shear-force generator structured for exerting a shear force to each volume element of the sample arranged in a container, a controller to control the shear-force generator to exert the shear-force of at maximum 10% of one shear-force intensity to each volume element of the sample arranged in the container, a light source, the beam path of which is directed into the container, an optical detector arranged to detect light emanating from the container, wherein the optical detector is a luminescence detector, a scatter light detector, or an absorbance detector, a computer including code to analyze light detected by the optical detector as a measure for the content of aggregated prion protein by comparing the light detected to a Reference Information Database containing pre-determined data relating detected light to content of aggregated conformation prion protein in relation to specific shear-force intensities and in relation to a diagnosis for a neurodegenerative disease pertaining to a reference sample used for generating the pre-determined data.

2. The device according to claim 1, wherein the Reference Information Database comprises data on amounts of aggregated conformation prion protein, which data are pre-determined for one specific shear-force intensity for a reference sample from a specific source of known diagnosis.

3. The device according to claim 1, wherein the code compares the data in the Reference Information Database to data representing amounts of aggregated conformation prion protein generated from an admixture of sample and native conformation prion protein, wherein the comparison of amounts of aggregated conformation prion protein is made for the same shear-force intensity.

4. The device according to claim 1, wherein the Reference Information Database comprises data for a specific neurodegenerative disease and/or a subtype thereof in association with pre-determined data on amounts of aggregated conformation prion protein, each generated at one of at least two, e.g. 3, 4, 12, 24, or 36 different shear-force intensities for each reference sample.

5. The device according to claim 1, comprising at least two containers, each arranged in a recess formed of a housing containing the optical detector and the light source, a thermostat and a lid section comprising a bearing for an axle carrying the rotor, wherein the housings are connected to one another and the lid sections are connected to one another.

6. The device according to claim 1, wherein the shear-force generator comprises a rotor arranged at a spacing from a stator, wherein the stator has extensions forming a funnel narrowing to an inlet which is coaxial to the rotor.

7. The device according to claim 1, wherein the shear-force generator has a stator formed of a portion of the wall of the container which for a portion of the circumference of the rotor is in parallel to the rotor at a constant spacing.

8. The device according to claim 1, employed for analysis for the presence of disease-related aggregated conformation prion protein in the biopsied mammalian sample by the steps of a) adding to the sample at least one native conformation prion protein, b) via the shear-force generator and controller, subjecting the mixture comprising the sample and at least one native conformation prion protein obtained in step a) to at least one shear-force intensity that is controlled to have a uniform intensity having an intensity range of maximally 20% of one shear-force value for a pre-determined number of cycles of a pre-determined time of shear-force acting and a pre-determined resting phase, c) with the code, following step b), determining the content of aggregated conformation prion protein for each of the shear-force intensities, characterized by d) comparing the content of aggregated conformation prion protein determined in step c) to pre-determined data on the content of aggregated conformation prion protein, which content was determined for native conformation prion protein subjected to the same shear-force intensity as in step b), wherein the pre-determined data on the content of aggregated conformation prion protein was determined in native conformation prion protein in admixture with a reference sample and these data are provided in Reference Information Database which in association with these data contains the neurodegenerative disease diagnosis for the patient from which the reference sample originates.

9. The device according to claim 8, wherein comprising, prior to step b) the mixture is divided into aliquots and in step b) at least two aliquots are subjected to a different shear-force intensity each and in step c) the content of aggregated conformation prion protein is determined for each aliquot and in step d) the content of aggregated conformation prion protein determined in step c) for each aliquot is compared to data on a pre-determined content of aggregated conformation prion protein.

10. The device according to claim 8, wherein comprising in step b) the mixture is subjected to a succession of at least two different shear-force intensities and the content of aggregated conformation prion protein is determined during or following subjecting the mixture to each one of the shear-force intensities.

11. The device according to claim 8, wherein comprising adding to the mixture at least one luminescent dye that is specific for the aggregated conformation prion protein prior to the step of subjecting the mixture to at least two different shear-force intensities and measuring the luminescence of the dye.

12. The device according to claim 8, wherein the device is configured for irradiating the mixture with light having a wavelength for exciting luminescence in the dye and measuring the luminescence of the dye during shear-force acting of step b) or during a resting phase of step b), without moving the volume occupied by the mixture relative a the shear-force generator generating the shear-force in step b).

13. The device according to claim 8, configured and controlled for irradiating the mixture with light having a wavelength that is scattered by the aggregated conformation prion protein and measuring scattered light exiting the admixture during step b), or during a pause of step b), with or without moving the volume occupied by the mixture relative a the shear-force generator generating the shear force in step b).

14. The device according to claim 8, wherein in step b) the rate of formation of aggregated conformation prion protein is determined from the content of aggregated state prion protein determined at the at least one shear-force intensity and the pre-determined data contain the rate of formation at the same shear-force intensity.

15. The device according to claim 8, wherein the content of aggregated conformation prion protein is determined as the time-resolved content and that the rate of formation of aggregated conformation prion protein is determined by non-linear regression analysis of an approximation on the determined time-resolved content of aggregated conformation prion protein for each of the shear-force intensities.

16. The device according to claim 8, wherein configured and controlled for adding at least one aggregated conformation prion protein to at least one aliquot of the mixture comprising the sample and at least one native conformation prion protein, wherein the at least one aggregated conformation prion protein is produced by subjecting a native conformation prion protein to a uniform shear-force controlled to an intensity range of maximally 1% of one shear-force intensity.

17. The device according to claim 8, wherein configured and controlled for treating in parallel at least one native conformation prion protein without addition of a sample of body fluid.

18. The device according to claim 1, wherein the Reference Information Database relates the medical diagnosis for a specific neurodegenerative disease and/or a subtype thereof in association with pre-determined data on amounts of aggregated conformation prion protein, which data were generated separately for each of at least two different shear-force intensities for each native conformation prion protein added to the sample.

19. The device according to claim 1, wherein the Reference Information Database includes data of the rate of formation of aggregated conformation prion protein for each of at least two different shear-force intensities, the rate of dissociation from aggregated conformation prion protein and/or the original amount of aggregated conformation prion protein in the sample.

20. The device according to claim 1, wherein the Reference Information Database contains data on pre-determined amounts which are specific for a single shear-force intensity and specific for a seed from a specific source.

21. The device according to claim 1, wherein the Reference Information Database contains the medical diagnosis for a specific neurodegenerative disease and/or a subtype thereof in association with the pre-determined data on amounts of aggregated conformation prion protein, each amount generated at one of at least two different shear-force intensities for each native conformation prion protein added and the diagnosis associated with the sample, and additionally contains the specific source or sample to which the diagnosis pertains.

22. The device according to claim 1, wherein the Reference Information Database contains information on the age of disease onset, sex of the mammal, duration of the disease, progression and/or severity of the disease.

Description

[0066] The invention is now described in greater detail by way of examples with reference to the figures that show in

[0067] FIGS. 1 to 6 fluorescence measurements specific for the amplification of the aggregated conformation of prion protein at different shear-force intensities for samples of known diagnosis,

[0068] FIGS. 7-10 fluorescence measurements specific for the amplification of the aggregated conformation of prion protein at different shear-force intensities for samples from post mortem brain samples,

[0069] FIG. 11 a schematic overview of an embodiment of the process,

[0070] FIG. 12 a schematic overview of an embodiment of the process,

[0071] FIG. 13 a schematic cross-section of a preferred device for use in the process,

[0072] FIG. 14 an exploded schematic view of the device of FIG. 13,

[0073] FIG. 15 a schematic view of a device,

[0074] FIG. 16 a schematic view of an embodiment of the device,

[0075] FIG. 17 a schematic cross-section along line A1 of FIG. 16, and

[0076] FIG. 18 a schematic cross-section along line A2 of FIG. 17.

EXAMPLE: AMPLIFICATION OF AGGREGATED STATE CONFORMATION AT DIFFERENT SHEAR-FORCE INTENSITIES

[0077] As samples, a 1:50 volume portion of brain homogenate was used and mixed with 2 mg/ml alpha-synuclein as the native conformation prion protein. The brain homogenate was prepared by homogenization in cold phosphate-buffered saline (PBS) containing 0.5% Triton X-100 and 1 complete EDTA-free protease inhibitor cocktail (Roche, Cat. No. 11873580001) using 20 strokes with a dounce homogenizer on ice. Brain samples were post-mortem from one healthy control (NEG.) and four different synucleopathy disease patients: Idiopathic Parkinson's Disease (IPD), Parkinson's Disease with Dementia (PDD), Dementia with LewyBodies (DLB), Muliple System Atrophy (MSA). The suspension was clarified by centrifugation at 2000g for 45 s, and the supernatant was used.

[0078] As another sample, 360 l human cerebrospinal fluid (CSF), stored at 80 C. were mixed with 40 l cold 10PBS containing 0.5% Triton X-100 and 1 complete EDTA-free protease inhibitor cocktail and clarified by centrifugation at 2000g for 45 s, giving the supernatant as human cerebrospinal fluid extract (CSFE). In this example the CSF sample was post-mortem from the same patient of Parkinson's disease with dementia (PDD) that was used above.

[0079] The native conformation prion protein was recombinantly expressed human alpha-synuclein of approx. 5.0 mg/ml water, stored at 80 C. and thawed at 37 C. at very mild agitation. The protein concentration was adjusted to 2.22 mg/ml using sterile-filtered water and 13.5 ml thereof were mixed with 1.5 ml of 10 concentrated PBS to yield 2.0 mg/ml human alpha-synuclein in PBS.

[0080] On ice, 15 ml of 2.0 mg/ml human alpha-synuclein in PBS were combined with 300 l brain homogenate supernatant, or alternatively with 300 l CSFE. Of this mixture, twelve identical aliquots of 1.2 ml each were filled into 1.5 ml sealed polypropylene test tubes (SureLock, Eppendorf). Into each test tube, one rotary shearing device having a rotor of 2.00 mm within a tube at a gap width of 0.30 mm was inserted. These assemblies were incubated at 37 C. for 15 min. Shear-force was applied by rotating the rotors with control of the rotating rate to at maximum 1% from the rate set for 5 s with a subsequent resting phase of 295 s for a total of 22 h. The rotating rates used are indicated in FIGS. 1 to 9. Generally, the device corresponded to WO 2012/110570.

[0081] Samples of 20 l were taken from each reaction mixture at time 0 h, 3 h, 6 h, 9 h, 12 h, and at 22 h under shear-force. The amplification of aggregated state conformation was determined by fluorescence spectroscopy of 15 l of the sample taken after mixing with 135 l Thioflavin T stock solution (30 M Thioflavin T in PBS buffer solution) with excitation at 450 nm (bandwidth 10 nm) and detection at 482 nm (bandwidth 20 nm).

[0082] The brain samples originated from patients diagnosed with the following Parkinson syndromes: Idiopathic Parkinson Disease (IPD), ICD-10: G20 [0083] Dementia in Parkinson's disease (PDD), ICD-10: G20, F02.3 [0084] Multisystem Atrophy (MSA), ICD-10: G90.3: [0085] Dementia with Lewybodies (DLB), ICD-10: G31.8, F02.3 [0086] Healthy Control (NEG.),
and the CSF sample originated from a patient diagnosed with [0087] Dementia in Parkinson's disease (PDD), ICD-10: G20, F02.3

[0088] For diagnosis, the ICD-10 codes (available at www.ICD-code.de, at http://apps.who.int/classifications/icd10/browse/2010/en#, at http://www.who.int/classifications/icd/en/, and at http://www.who.int/classifications/icd/en/GRNBOOK.pdf) were used.

[0089] In detail, F00 Dementia in Alzheimer's disease: Alzheimer disease (AD) is a primary degenerative cerebral disease of unknown etiology with characteristic neuropathological and neurochemical features. The disorder is usually insidious in onset and develops slowly but steadily over a period of several years. Associated with the deposition of seed of abeta protein, tau protein, and sometimes synuclein protein [0090] F00.0*, G30.0*: Dementia in Alzheimer's disease with early onset. Dementia in Alzheimer disease with onset before the age of 65, with a relatively rapid deteriorating course and with marked multiple disorders of the higher cortical functions. Includes: (i) Alzheimer disease, type 2; (ii) Presenile dementia, Alzheimer type; (iii) Primary degenerative dementia of the Alzheimer type, presenile onset. [0091] F00.1*, G30.1*: Dementia in Alzheimer's disease with late onset. Dementia in Alzheimer disease with onset after the age of 65, usually in the late 70 s or thereafter, with a slow progression, and with memory impairment as the principal feature. Includes (i) Alzheimer disease, type 1; (ii) Primary degenerative dementia of the Alzheimer type, senile onset; (iii) Senile dementia, Alzheimer type. [0092] F00.2*, G30.8*: Dementia in Alzheimer's disease, atypical or mixed type. Atypical dementia, Alzheimer type [0093] F00.8, G30.9: Dementia in Alzheimer's disease, unspecified

[0094] F02 Dementia in other diseases classified elsewhere. Cases of dementia due, or presumed to be due, to causes other than Alzheimer disease or cerebrovascular disease. Onset may be at any time in life. [0095] F02.0*, G31.0*: Dementia in Pick's disease (Frontotemporal lobular Dementia, FTD). A progressive dementia, commencing in middle age, characterized by early, slowly progressing changes of character and social deterioration, followed by impairment of intellect, memory, and language functions, with apathy, euphoria and, occasionally, extrapyramidal phenomena. [0096] F02.2*, G10*: Dementia in Huntington's disease (HD). A dementia occurring as part of a widespread degeneration of the brain. The disorder is transmitted by a single autosomal dominant gene. Symptoms typically emerge in the third and fourth decade. Progression is slow, leading to death usually within 10 to 15 years. Includes: Dementia in Huntington chorea [0097] F02.3*, G20*: Dementia in Parkinson's disease (PDD): dementia developing in the course of established Parkinson disease. No particular distinguishing clinical features have yet been demonstrated. Includes (i) Hemiparkinsonism, (ii) Paralysis agitans, (iii) Parkinsonism or Parkinson disease (NOS (not otherwise specified), idiopathic, primary) [0098] F02.3*, G31.82: Lewy body Dementia (DLB), Lewy Body Disease (LBD). A progressive degenerative dementia. Persons with LBD will show markedly fluctuating cognition. Persistent or recurring visual hallucinations with vivid and detailed pictures are often an early diagnostic symptom.

[0099] A81 Atypical virus infections of central nervous system. Prion diseases of the central nervous system. [0100] A81.0*, F02.1*. Dementia in Creutzfeldt-Jakob disease. A progressive dementia with extensive neurological signs, due to specific neuropathological changes that are presumed to be caused by a transmissible agent. Onset is usually in middle or later life, but may be at any adult age. The course is subacute, leading to death within one to two years. [0101] A81.8: Other atypical virus infections of central nervous system: Kuru [0102] A81.9: Atypical virus infection of central nervous system, unspecified: Prion disease of central nervous system.

Amyloidosis:

[0103] 168.0* Cerebral amyloid angiopathy (E85.-+)

[0104] [Possibly to be Extended]

Parkinson Syndromes:

[0105] G20: Idiopathic Parkinson Disease (IPD) [0106] G20, F02.3: Dementia in Parkinson's disease (PDD) [0107] G90.3: Multisystem Atrophy (MSA) [0108] G31.8, F02.3: Dementia with Lewybodies (DLB)

Motor Neuron Disease:

[0109] G12.2: Motor neuron disease: includes (i) Familial motor neuron disease and (ii) Amyotrophic Lateral sclerosis (ALS).

[0110] FIGS. 1-6 show the fluorescence detected in 3 independent repetitions of the examples. In FIG. 1, the sample was BN449-PDD, in FIG. 2 BN379-IPD, in FIG. 3 BN175-MSA, in FIG. 4 BN526-DLB, in FIG. 5 BN449-PDD total CSF, and in FIG. 6 BN276 Healthy Control.

[0111] The results show that in the healthy control (NEG.), no aggregated conformation prion protein was generated. All the samples from the patients diagnosed with Parkinson syndromes resulted in the generation of amplification that was dependent on the shear-force intensity (rotation rate, application time, resting time and cycle number) and dependent on the origin of the sample. The process was highly reproducible in three independent experiments performed on the same brain tissues.

[0112] In FIGS. 1-10, the shear-force intensities are given as rpm of the rotary shear-force generator on the X-axis, the amounts of aggregated conformation prion protein detected are given on the Y-axis with the individual curves for given for the time points indicated on the right (h application of shear-force intensities).

[0113] In FIGS. 1-4 and 6, the contents of aggregated conformation prion protein generated at a pre-determined application of one shear-force intensity each is depicted, showing a specific pattern of amplification for each sample for the shear-force intensities. In FIG. 5, the contents of aggregated conformation prion protein at 0 h, and generated at 3 h, 9 h and 12 h, respectively, are depicted, showing the different rates of amplification at different shear-force intensities. For the process of the invention it is therefore generally preferred that the shear-force intensity is pre-determined and the same for the sample and for pre-determined contents, e.g. the shear-force intensity can generally be pre-determined for shear-force applied, duration of shear-force application, duration of resting phase for each cycle, and repetition number of cycles.

[0114] The results show that the process of the invention differentiates between samples of different pathologies and between subtypes, e.g. specific disease presentation of individual patients.

[0115] FIGS. 7-9 show results for the same process conditions for post mortem brain (BN) samples, wherein numbers designate individual samples. In contrast to the samples of FIGS. 1-6, the disease status of patients from whom the samples of FIGS. 7-9 originate is unknown to the persons involved in performing the analytical process. In these Figures, the content of aggregated prion protein generated at different time points is indicated.

[0116] The results depicted in FIGS. 7-9 show that the amplification greatly varies between samples and between points in time of shear-force application.

[0117] FIG. 10 shows the amount of aggregated conformation prion protein from an admixture of a post mortem sample of a brain histologically determined as Alzheimer and human -synuclein as the native conformation prion protein. The result shows that the Alzheimer sample which was diagnosed to contain A and tau protein aggregates when subjected to specific shear-force intensities did not significantly induce the generation of aggregated conformation prion protein from the native conformation -synuclein. This results demonstrates that at least for this Alzheimer sample, generation of aggregated conformation prion protein from a native conformation prion protein is specific for the sample.

[0118] A comparison of the amplification of aggregated prion protein at single shear-force intensities, i.e. for different rotation rates at the same cycles for a total of 22 h allows to identify similar patterns of amplification, preferably an identification of the unknown sample according to similarities of the amplification pattern generated from a sample of known diagnosis. In this process, the sample of known diagnosis serves as a reference sample. In detail, the samples of FIGS. 8 and 9 show a similar amplification pattern at specific shear-force intensities (22 h) as the IPD sample of FIG. 2.

[0119] Therefore, it is assumed that the process can differentiate samples according to the progression of accumulation of aggregated prion protein during disease.

[0120] FIG. 11 schematically shows an overview of the process for analysis. Reference samples, e.g. post mortem brain tissue samples, serum, CSF or urine, each in association with the specific diagnosis for a neurodegenerative disease are provided as a library of reference seeds (Reference Seed Library). In the library, the reference samples can be assigned to the respective prion protein, represented by alpha-synuclein (-Syn), tau (Tau) or amyloid beta (A). For the reference samples, pre-determined data on the amounts of aggregated conformation prion protein is generated by application of specific shear-force intensities (SSA) from an admixture containing reference sample and native conformation prion protein. The measurement can be by optical determination of the amounts of aggregated conformation prion protein in Western blots or using an optical detector receiving irradiation from the admixture. As indicated by the double arrows between the Reference Seed Library and the databank Reference Information Database, the data on the detected amounts (Amplification Profiles) of aggregated conformation prion protein in respect of each shear-force intensity are stored in a computer and stored in a databank (Reference Seed Library) in association with the respective specific diagnosis (Disease, IPD, PDD, MSA, DLB, FTD, AD), preferably including at least one subtype (G20, F02.3, G90.3, G31.82, G31.0, F00.0, G30.0, G30.1, G30.8, G30.9) according to a classification (ICD10). The databank (Reference Information Database) therefore for each reference sample (Sample Information) in association with the specific disease, preferably its subtype, for the native conformation prion protein used (A, Tau, -Syn) contains the amounts of aggregated conformation prion protein for each shear-force intensity (Amplification Profiles).

[0121] As indicated by the double arrows between the Diagnostic Process and the Reference Seed Library, patient samples can be integrated into the reference samples (Reference Seed Library) once the diagnosis associated with the sample is known.

[0122] As generally preferred, the sample to be analysed (Patient Sample) in admixture with the same native conformation prion protein (separated admixtures for each of -Syn, Tau and A) as at least one reference sample of the Reference Seed Library is subjected to the same at least one shear-force intensity (SSA), and amounts of aggregated conformation prion protein are measured. Generally preferred, the sample to be analysed is of the same type as the reference sample, e.g. blood serum, lymph fluid, urine, CSF or a tissue sample.

[0123] The amount of aggregated conformation prion protein generated at specific shear-force intensities generated for a sample (Patient sample) is compared to the amount of aggregated conformation prion protein generated at the same shear-force intensities for the same native conformation prion protein (A, Tau, -Syn) each (Computer), allowing the identification (Diagnosis) of the diagnosis associated to the reference sample in the databank (Reference Information Database) by this comparison.

[0124] FIG. 12 in addition to FIG. 11 shows that preferably for each application of a shear-force intensity in addition to a sample to be analysed (Patient), native prion protein only, i.e. without a seed (negative Control) and a reference sample (Reference Seed) as a positive control can optionally be used in the process in parallel to the sample. The reference sample can e.g. be taken from the Reference Seed Library. Generally, the reference sample can be an aggregated conformation prion protein produced by application of one shear-force intensity to an admixture of one patient sample of known disease with native conformation prion protein, preferably followed by at least one further application of the same shear-force intensity to an admixture of an aliquot of the resultant product with the same native conformation prion protein.

[0125] Further, FIG. 12 shows that preferably for each application of a shear-force intensity to a sample or reference sample, at least one of the following data is stored: the specific shear-force generator or its drive D, preferably including the frequency generated by its drive D, the type of lid L, the type of sample compartment S, e.g. comprising the lid L, the thermostat T and the optical detector O, the current temperature and/or the specific temperature control element T and/or the type and/or specific optical detector O, including the optical measurement data are stored, e.g. using an interface (Computer Interface) coupled to each shear-force generator of the device and coupled to the computer (Computer) for transmitting the data. As further preferred, FIG. 12 shows that for measuring the amount of aggregated conformation prion protein, the computer is set up to store the temperature, time course (Timing, t), the shear-force generator (Drive, D), and the optical detector (Optics, O) for each application of a shear-force intensity, preferably including storing the time course of the detected amounts of aggregated conformation prion protein, e.g. in the form of amplification profiles for each native conformation prion protein in an admixture using e.g. a programme for storing these data (Device Software). Further, the device optionally comprises a programme (Evaluation Software) for generating from these data which are measured and stored during the application of at least one shear-force intensity the amounts of aggregated conformation prion protein, each for the combination of specific native conformation prion protein of the admixture (A, Tau, -Syn), with the measured values for shear-force intensity, temperature, time-course of the application of shear-force intensity, specific shear-force generator and/or specific optics, e. g. detector. The evaluation software has access to the databank. The device contains or has access to the databank comprising pre-determined amounts of aggregated conformation prion protein generated at at least one shear-force intensity for reference samples (Reference Information Database), and the computer is set up to compare the amplification profiles generated for samples for each shear-force intensity (double arrows between Computer and Reference Information Database), allowing the association of a disease, preferably including its subtype, stored for a reference sample to the sample analysed. The computer is optionally set up to edit this diagnosis (Diagnosis).

[0126] FIGS. 13 and 14 show cross-sections of a preferred shear-force generator for use in the invention. By way of a connection 1, e.g. a data transfer line to a computer (not shown), a drive control unit 2 is controlled by the computer. The drive control unit 2 controls the drive motor 3 to a uniform rotation frequency. The drive control unit 2 and the drive motor 3 can also be termed drive D. The rotor 9 is arranged on an axle 9a that is connected to the drive motor 3 by a coupling 4, which preferably is a magnetic coupling. The rotor 9 is arranged within a container 8 and a stator 10 is arranged between the rotor 9 and the container 8. The stator 10 is arranged with a spacing to rotor 9, which preferably is a uniform spacing to the radial outer surface of rotor 9, e.g. forming a channel of ring-shaped cross-section, and stator 10 is arranged with a spacing to container 8. Preferably, stator 10 is mounted on the lid section L, also containing a bearing for axle 9a. Optionally, lid section L contains drive motor 3 and drive control unit 2. Stator 10 preferably has extensions 10e opposite the axle 9a carrying rotor 9, which extensions 10e form a funnel having an inlet 10i and reducing the free inner volume of container 8 in the region between the end of rotor 9 opposite the axle 9a and container 8. The inlet 10i of the stator 10 is preferably arranged coaxially to rotor 10 and forms the narrow exit of the funnel formed by extensions 10e, guiding liquid to the front surface section of rotor 9, allowing a pumping action by rotor 9 to move liquid into the spacing between the radial surface of rotor 9 and stator 10. Opposite the inlet 10i, the outlet opening 10o has at least the cross-section of the spacing between rotor 9 and stator 10.

[0127] The container 8 provided for receiving a sample 20 is arranged within a housing 7, which preferably at least sectionally is a thermostat T, preferably having form fit to the container 8. Preferably, the thermostat T for each container 8 has a temperature sensor and is independently computer-controlled. The open end of the container 8 is closed by a lid 6 and a seal 5. As shown, the section of housing 7 embracing the section of container 8 between extensions 10e of stator 10 and the bottom of the container 8 opposite its opening is provided with a light source 17 arranged to irradiate the inner volume of the container 8 and an optical detector 14 arranged to receive irradiation exiting the inner volume of the container 8, wherein preferably the beam path of the light source 17 crosses the beam path of the optical detector 14 in the area between the inlet 10i and the bottom of the container 8. The beam path 19 generated by the light source 17 can cross the exiting beam path 11 towards the optical detector 14 e.g. at an angle of 90. Both the light source 17 and the optical detector 14 are coupled to a computer for control of the light source 17 and for receiving measurement signals from the detector 14. In the exiting beam path 11, a wavelength discriminator 13, e.g. an optical filter can be arranged. In the beam path 19 generated by the light source 17, a wavelength discriminator 18, e.g. an optical filter can be arranged. The housing 12 for the detector 14 and/or for the light source 17 has a dataline 16 for transmitting data on irradiation and measurement signals to a computer. Optionally, a control unit 15 for controlling the thermostat T and/or the light source 17 is arranged at the housing 12.

[0128] Preferably, housing 12 containing the light source 17, optionally provided with a wavelength discriminator 18, and optical detector 14, optionally provided with a wavelength discriminator 13, form an integrated optical unit O. The optical unit O can be mounted releasably to thermostat T and to adjacent lid section L, wherein these elements form a recess for receiving a portion of container 8.

[0129] The scale indicated in FIGS. 13 and 14 is an exemplary scale, indicating that preferably each container 8 is provided with an individual controlled shear-force generator comprising a rotor 10 within a stator 9, a thermostat T, a light source 17 and a detector 14 and a controlled drive motor 3 within a scale of 7 to 12 mm, preferably 9 mm, for arrangement in a row or grid. FIG. 15 shows a generally preferred arrangement of at least two, e.g. of 8 or 12 containers that are connected to one another, each provided with a separate shear-force generator comprising a drive D and a lid section L, wherein drives D and lid sections L as well as thermostats T and optical units O, respectively are connected to one another in the same spacing for arrangement around spaced-apart coupled containers 8.

[0130] FIG. 16 shows another embodiment, wherein the shear-force generator is formed of a rotor 9 and stator 10 consisting of a wall section of the container 8. The wall section of the container 8 forming the stator 10 is e.g. arranged for a portion, e.g. at least 1 to 270, e.g. for 30 to 180 about the circumference of the rotor 9 at a constant distance. As shown in FIG. 16, the rotor 9 is preferably arranged asymmetrically within container 8, leaving a free inner volume section for optical detection.

[0131] The container 8 can e.g. have a circular, oval or egg-shaped cross-section, and the rotor 9 can be arranged within a part of the cross-section having a smaller or larger diameter.

[0132] FIGS. 17 and 18 show that the container 8 preferably forms a stator 10 arranged at a constant distance from the rotor 9 for a portion about the circumference of the rotor 9, wherein this distance forms the smallest passage between rotor 9 and stator 10. Accordingly, the surface of rotor 9 preferably is parallel to the stator 10 for a portion of the circumference of the rotor 9.

REFERENCE NUMERALS

[0133] 1 connection [0134] 2 drive control unit [0135] 3 drive motor [0136] 4 coupling [0137] 5 seal [0138] 6 lid [0139] 7 housing [0140] 8 container [0141] 9 rotor [0142] 9a axle [0143] 10 stator [0144] 10e extensions [0145] 10o outlet opening [0146] 10i inlet [0147] 11 exiting light path [0148] 12 housing containing optical detector and [0149] light source [0150] 13 wavelength discriminator [0151] 14 optical detector [0152] 15 control unit [0153] 16 dataline [0154] 17 light source [0155] 18 wavelength discriminator [0156] 19 beam path from light source [0157] 20 sample [0158] D drive [0159] L lid section [0160] T thermostat [0161] S sample compartment [0162] O optical unit