GB2400669A

GB2400669A - Determining haematological parameters and indicators by harnessing the dynamics of erythrocyte

Info

Publication number: GB2400669A
Application number: GB0308926A
Authority: GB
Inventors: Christopher Barnes
Original assignee: Individual
Current assignee: Individual
Priority date: 2003-04-17
Filing date: 2003-04-17
Publication date: 2004-10-20
Anticipated expiration: 2023-04-17
Also published as: GB0308926D0; GB2400669B

Abstract

A method and device to harness the dynamic properties of aggregating erythrocytes and other blood aggregations in vitro or in vivo to indicate a haematological parameter comprises, mixing a blood sample to a predefined rate of shear or for a predetermined mixing time if in vitro, or halting or reducing blood flow for a predetermined duration if in vivo, selecting a defined region in the temporal evolution of the aggregation, making at least two time successive remote examinations to yield quantities which reflect the aggregation state, and mathematically manipulating said quantities together to correlate with or indicate the haematological parameter. A capacitance measuring system measures dielectric properties of the blood samples by applying a frequency (A1) to a sample cell.

Description

A method and device to obtain haematological parameters and indicators by

means of harnessing the dynamics of erythrocyte and other aggregation.

This in invention relates to a method and device to harness or employ the dynamic behaviour of specifically discovered and defined temporal phases of erythrocyte and other aggregation to obtain haematological parameters and indicators therefrom.

The background technical field ofthis present invention impinges on several areas of Science. For instance,it has been long documented that blood Is a dielectric material There have been several academic papers claiming to relate static blood properties Particularly those of red cells of various species to their manifest dielectric properties such as for example cell membrane capacitance, plasma conductivity and cytoplasm conductivity. The results for these parameters as published are often very inconsistent with each other and it is the opinion of the present inventor that this is because the hemodynamc status of these various bloods has often not been taken into account.

Several patent specifications that claim to be able to measure static red cell parameters by dielectric and /or magnetic interaction with blood and haemoglobin.

As yet however there appears to be little if any successful instrumentation havmg reached the market place. Three patents appear to disclose dielectric means of measuring erythrocyte sedimentation rate but the various owners of those rights would appear not to have exploited those technologies. It Is the present view of the present inventor that such technologies have not been successfully exploited because of the overwhelming effects of electrode polarization, if electrodes are dipped into the blood, or to stray capacitance problems,container size errors and hemodynamic effects in the cases where contactless measurements have been attempted.

The approach taken by the present inventor and Invention is quite different and involves an interpretation and measurement of the causal effects of erythrocyte and other particulate aggregation. It has recently been suggested that transient erythrocyte aggregation gives rise to at least some ofthe backscattered ultrasonic signal in ultrasonic blood flow investigations (Sennaoui et al, University of Paris) and likewise gives rise to some of the amplitude effect in standard optical and infra-red pulse oximetery systems.

It has further been suggested without firm conclusion that erythrocyte aggregation may modify the apparent measured red cell membrane capacitance by up to a factor ofthree in stationary whole blood but that this effect can be removed by hemodynamic shear or when insufflcent plasma and/ or fibrinogen (the main blood clotting protein) is present (Beving et al European Biophys. J. (1994) 23;pp 207 -215) Traditionally whole blood (erythrocyte) aggregation and that of other blood components has been studied in vitro by optical aggregometers and direct current conductivity systems wherein pairs of electrodes or disposable electrodes directly contact the sample under investigation. In the case of whole blood two specific points on the aggregation curve have traditionally been recorded at 10 and 20 seconds into each aggregation, the so called tlO and t20 points. In certain pathologies the aggregation rate can be seriously disturbed in either direction from the normal range. Specialised optical aggregometer systems have also been employed for pseudo in vivo use such as for example in the flow circuits of kidney dialysis machines as an Indicator of drastic circulatory problems.

Using such systems under experimental conditions it has been possible to point out that with feline blood the rate at It O or t20 may sometimes correlate with the cat haematocrit over a limited range. The present inventor has privately experimented with and researched the aggregation of human and equine erythrocytes in a variety of anticoagulation media and with a variety of different kinds of energy probe to explore their temporal dynamic behaviour after known quantities of mixing (shear) . It has been previously stated that aggregation time curves have a simple mathematical form,being predominantly exponential growths.

The present inventor does not share this view nor is it consistent with the observations of the present inventor or the physico- chemical processes involved. According to the present inventor and invention there are at least four recognisable specific temporal phases in the time evolution of the dynamic behaviour of erythrocyte aggregation which generally but not exclusively proceed for a period of 20 -30 seconds to a few minutes after mixing for a predetermined time or with a predefined rate of shear and then stopping the said mixing and vertically placing the blood container or after similarly halting or reducing m vitro or in Volvo flow or the onset of stasis starting with a beginning phase, during which shear energy is dissipated by interracial slip and similar processes governed partly but not exclusively by plasma viscosity and protein content, especially fibrinogen and followed about 5 seconds later generally but not exclusively by an attractive phase governed mainly by but not exclusively by the blood hematocrit concentration, followed generally but not exclusively about 10 seconds later by a repulsive phase lasting about 10 seconds governed mainly but not exclusively by the blood cell mean cell volume, followed by a quasi stable phase lasting several minutes.After the quasi-stable phase the aggregated erythrocytes visibly begin to sediment under gravity if suitably contained in a vertical tube. An exception to this is if the system is not anticoagulated,when the additional phase of clotting will occur or when specific clotting factors are added.

Another exception is when specific antibody -antigen type reactions are allowed to occur as in blood typing studies when the final stage of sedimentation will be arrested or when blood components cause the aggregation or agglutination of a secondary component system such as latex or microspheres as in infectious disease detection drug and/or delivery evaluation which can be similarly studied according to the present invention. Further according to the present inventor and invention an explanation is offered for the behaviour of the attractive phase as follows there exists electrostatic forces dependent on the biochemistry of fibrinogen certain other plasma proteins and the surface proteins and charge ofthe erythrocytes which pull the erythrocytes towards each other.Further according to the present invention the fewer erythrocytes there were to start with the more well dispersed they are in the plasma so the larger relative physical change they make when they aggregate thus preferably remote measurement of rate of the aggregation here gives a measure of the volume occupied by the erythrocytes the so called packed cell volume or haematocrit.

Further and advantageously according to the present invention when the erythrocytes get too close together their approach is limited in the newly reported and so called repulsive phase by their shape or size and in healthy samples since this shape is a fixed form of ellipsoid then this approach Is governed simply by the mean cell volume. Thus remote measurement of the temporal change here can yield a measure or estimate of mean cel I -- 4- volume In the quasi stable phase minimal temporal change allows Depending on the energy probe used, an alternative measure ofthe haematocrit and/or the measure the cytoplasmic content of the erythrocytes which is haemoglobin. Further according to the present invention, contrasting measurements of the haematocrit values obtained from the attractive and quasi stable phases ofthe aggregation can be used as an indicator of certain pathologies or disease states.

Those skilled in the art will appreciate that m non -healthy samples and in the case of those with certain pathologies, the rates Degrees and onset point(S) of identified phases of aggregation according to the present invention may be disturbed and thus comparison of haematological parameters measured according to the method herein with the same as measured by other more traditional techniques can also itself be a powerful diagnostic tool for certain disease states, such diseases Include SLE (lupus), diabetes, Sickle Cell, Malaria and several others. Further according to the present inventor and invention because of the physical size of the aggregates at the differently defined phases of the aggregation as defined by the present inventor and invention, different energy probes can be employed with optimal wave types Energies and frequencies to probe each said phase and said temporal dynamic behaviour thereof.

According then to the first specific embodiment of the present invention there is provided a method to employ or harness the dynamic behaviour of specific temporal phases of the aggregation of erythrocytes and /or blood components and blood related aggregations or agglutinations, which said dynamic behaviour,its state or rate or degree or time -dependent onset point(s) allowing resultant determination of at least one haematological parameter or two or more such parameters which may be one(s) with a numeric value such as those of erythrocytes or clotting function or an indicator such as blood type or the presence or absence of disease state comprising a first step of selecting a defined region In the temporal evolution of moving aggregating erythrocytes or other such blood or blood related components undergoing aggregation or agglutination and comprising a second step making at least two time successive(non-simultaneous) remote preferably non-optical examinations thereofto yield real or virtual quantities which reflect the magnitude of which reflect the rate of or state of or degree of completeness of the said aggregation and further comprising a third step wherein the said quantities or their magnitudes are manipulated together mathematically so as to measure, determine or correlate with the said haematological parameter(s) or yield the said haematological indicator.

Further according to the first said embodiment the said remote examinations can be either in vivo or in vitro. Further according to the first said embodiment the means of remote examination may involve the supply of energy from a source and evaluation of energy loss and/or scattering or reflection by a probe Further said source and probe may involve the supply and detection of shear or longitudinal compression wave energy of any frequency or waveform or any simultaneous or sequential combination of frequencies and waveform. Further and as an alternative to compression-wave energy the said source and probe measurement may involve the supply and detection of transverse wave electromagnetic energy of any known polarization and with a single constant exciting frequency when making the said two or measurements referred to herein above or may mvolve the use of a separate single but different (preferably but not exclusively escalated) exciting frequency at each of the two or more said time successive measurements Further as a said means of remote examination and involving an energy loss evaluation probe, two or more time successive (sequential) measurements of the electrical capacitance of the system as a whole at fixed or differing (preferably escalating) frequency, preferably within the range I kHz to 3 GHz and preferably in accordance with a prior art method for capacitance measurement as previously disclosed by the present Inventor in the disclosed priority document GB 2336685 of the present inventor. Further according to the first specific embodiment above the said parameter may be the haematocrit. The said parameter may further be the mean cell volume. The said parameter may further relate to the cytoplasmic content of the erythrocytes Further according to the present invention those skilled in the art will appreciate that by calculation from two ofthe above said parameters it is possible to obtain an extra said parameter, the so called red cell count within the scope ofthe claims herein. Further according to the present invention, the said blood parameter or indicator may relate to clotting function blood type or disease state or diagnosis thereof.

According then to the second specific embodiment of the present invention there is provided apparatus to employ or harness the dynamic behaviour of specific temporal phases of erythrocyte and/or other particulate aggregation or agglutination in blood /blood components or blood related systems m vitro or in vivo such said employment of or harnessing of such said dynamic behaviour its rate or state or degree or onset point (s)of allowing resultant determination of at least one or two or more related haematological parameter(s) which may be one(s) with a numeric value such as those of erythrocyte volume number density and form factor or of clotting function or of an indicator such as blood type or the presence or absence of disease state or infectious disease state comprising a means to mix blood for predetermined time or at predefined rate of shear and of stopping the said mixing if in vitro or means to halt or reduce flow for predetermined time if in vivo also with means of selecting a defined time region m the temporal evolution of moving aggregating erythrocytes or particles of other such blood or blood related components undergoing aggregation or agglutination and comprising a means of making at least two time successive (non -simultaneous) remote preferably non- optical examinations thereof to yield real or virtual measurement quantities which reflect the magnitude of or which reflect the state of or rate of or degree of completeness of the said aggregation and further comprising a means wherein the said quantities or their magnitudes are manipulated together mathematically preferably though not exclusively by division or subtraction of amplitudes or phases so as to measure or correlate by means of linear or other algorithm with the said haematological parameter(s) or yield the said haematological indicator. Further according to the second said specific embodiment of this present invention the said apparatus is provided with either means of making the said determination either in vivo or in vitro. Further according to the second said embodiment the means of examinations may mvolve the use of an energy source and an energy probe for the detection of energy loss Scattering or reflection and may further involve the supply and detection of shear or longitudinal (compression) wave energy of any frequency or waveform or any simultaneous or sequential combmaton of frequencies and waveform. Further according to the said second specific embodiment as an alternative to shear or longitudinal wave energy the said remote means of examination may involve the supply and detection may be of transverse wave (electromagnetic) energy of any known polarisation and with a single constant exciting frequency when making the said two or measurements referred to herem above or may involve the use of a separate single but different (preferably but not exclusively escalated) exciting frequency at each of the two or more said time successive measurements' each said exciting frequency being preferably within the range 1 kHz to 3 GHz Further according to the present second specific embodiment the said remote means of examination may involve two or more time successive measurements of the electrical capacitance of the system as a whole, in accordance with a a prior art method and apparatus for capacitance measurement as previously disclosed by the present inventor in priority document GB 2336685 Further according to the second specific embodiment the said at least one parameter may be the haematocrit.

Further additional and /or alternative said parameters may be or relate to the mean cell volume, or relate to the cytoplasmic content ofthe erythrocytes. Further according to the present nvention those skilled in the art will appreciate that by calculation from two of the above said parameters it is possible to obtam an extra said parameter, the so called red cell count and that the apparatus may be provided with means to do this within the scope of the claims herein Further according to this second embodiment of the present invention, the said blood parameter or indicator may relate to clotting function, blood type or disease state or diagnosis thereof The present invention is now described in more detail by way of reference to the drawing in which figure 1 'illustrates the measurement of aggregation graphically by way of a preferred but not exclusive example of a (capacitance) measurement and further shows the various regions and typical time zones of interest.

Figure 2 shows the perceived pictorial behaviour of initially dispersed erythrocytes before and after aggregation.

Figure 3 shows a block diagram of part of the preferred apparatus scheme for parameter measurement m Volvo In figure 3 1 is the exciting wave energy with a preferably continuous amplitude Al, 2 is the sample and remote transducer arrangement in a state of aggregation at time tl, 3 is the same or similar at a different stage of aggregation at time t2., 4 and 5 is a sample and hold gate holding respectively amplitudes (or phases) A2 which is a function of the system at time tl and AS which Is a similar function of the system at a later measurement time t2. 6 is a computation circuit which preferably in its simplest form may be a divider or subtracter followed by a linear algorithm to drive the output display or recording: device 7. Those skilled in the art will recluse that more sets of blocks such as pairs 2 and i 4 and 3 and 5 may be added to the system within the scope of the present claims and will I likewise reahse that there are many different electronic circuit and computational variations that can be used to achieve the effect described all of which when used as such lie within the scope of the present claims It will be further realised by those skilled I in the art that the parts of the apparatus described in 2 and 3 can be modified and housed within a finger,vein, limb,or other bodily part clamp or restraining device to give the in veto version according to the present claims The present inventor in private study has found that an initially mixed blood sample when subjected to manual or automatic 2 dielectric scanning or impedance analysis over finite times will yield an apparent non relaxatonal or resonance type behavour in the early stages ofthe scan. Some academic parties have observed similar effects and tried to ascribe a multiple relaxation or energy coherent effects in cell membranes It is disclosed here that a possible cause of this observed process is simply down to the newly discovered significance of erythrocyte and other aggregation behavour and the specific temporal phases thereof as defined and disclosed by the present inventor and invention and the way in which the temporal variation of the primarily but not exclusively the electrical capacitance interacts with the time sequence of the impedance scanning measurement. An extreme example of this according to the method of and with the apparatus of the present invention is I when the said number of remote examinations according to the present claims herein is very high or at a very fast rate according to the present invention. Thus the findings of -1 1 the present inventor and use of the present invention according to one of its embodiments can thus account for these previous uncertain observations.

It should be stressed according to this present invention that when the method and I apparatus are used in accordance with the non-optical aspects of the invention as are preferable it is particularly advantageous when used for in vitro measurements of samples in sample tubes surrounded wholly or partly by patient sample 1/D labels I because the energy source and probe according to the present invention can penetrate such labels which may not necessarily be the case for optical sources. Likewise and similarly in the in vivo case, non-optical energy sources can be arranged to penetrate human or animal skin and even painted nails!

1. A method to employ or harness the dynam c behaviour of specific temporal phases of the aggregation of erythrocytes and /or blood components and blood related aggregations or agglutinations in vitro or in vivo, which said dynamic behaviour,ts state or rate or degree or onset point (s) of allowing resultant determination of at least one haematological parameter which may be one with a numeric value such as those of erythrocytes or clotting function or an indicator such as blood type or the presence or absence of disease state comprising a first step if in vitro of mixing the blood to a predefined rate of shear or for a predetermined mixing time and then stopping the said mixing or if m vivo halting or reducing its flow for a predetermined duration, comprising a second step selecting a defined region in the temporal evolution of moving aggregating erythrocytes or other such blood or blood related components undergoing aggregation or agglutination and comprising a third step making at least two time successive(non-smultaneous) remote examinations thereofto yield real or virtual quantities which reflect the magnitude of which reflect the rate of or state of or degree of completeness of the said aggregation and further comprising a final step wherein the said quantities or their magnitudes are manipulated together mathematically so as to measure Determine or correlate with the said haematological parameter(s) or yield the said haematological indicator, either in vivo or in vitro. 1'

2. Method as in claim 1 where there is determination of two or more haematological parameters and wherein said remote examinations are preferably non-optical and involve the supply of energy from a source which can penetrate patient l/D labels or skin or nail and evaluation by a probe.

3. Method as in claim 2 wherein said probe measures energy loss.

4.Method as in claim 2 or 3 wherein said probe measures reflection 5. Method as m any of claims 2-4 wherein said probe measures scattering 6. Method according to claim 2 above wherein the means of remote physical measurement may involve the supply and detection of shear or longitudinal (compression) -wave energy of any frequency or waveform or any simultaneous or sequential combination of frequencies and waveform 7 Method according to the said claim 2 above wherein the said remote means of physical measurement involves the supply and detection of transverse wave electromagnetic energy of any known polarization and with a single constant exciting frequency when making the said two or measurements referred to herein above )k 8 Method as in claims 2 and 7 above but involving the use of a separate single but different (preferably but not exclusively escalated) exciting frequency at each of the two or more said time successive measurements 9 Method according to claim 1 above wherein the said remote examinations involve two or more time successive (sequential) measurements of the electrical capacitance of the system as a whole at fixed frequency, preferably within the range I kHz to 3 GHz.

Method according to claims 1 and 9 wherein said time successive measurements involve measurement of electrical capacitance at different successive preferably escalating frequencies.

11 Method as in either claim 9 or 10 wherein said capacitance measurement is in accordance with the prior art method for capacitance measurement as previously described by the present inventor in the disclosed priority document GB 2336685 12. Method according to any of claims 1-1 1 above wherein above one of said parameters may be the haematocat, measured preferably in the so called attractive phase of the aggregation as defined by the present inventor and disclosed by the present invention at preferably some 5-10 seconds aftemnxmg or halting of flow.

IS

13 Method according to any of claims 1-12 above wherein one of said parameters may be the mean cell volume measured preferably in the repulsive phase of the aggregation and where erythrocyte approach is limited as defined by the present inventor and disclosed by the present invention.

14. Method as m any of claims 1-13 above wherem one of said parameters relates to either the haematocrit or the cytoplasmic content which is preferably measured in the quasi -stable phase of the aggregation as defined by the present inventor.

15. Method according to any of claims 1-14 wherein one ofthe said parameters is the red cell count.

16 Method according to any of claimsl -I l above wherein the said parameter or I indicator relates to clothng function 17. Method as In any of claims 1-11 above wherein the said indicator relates to blood type.

18. Method as in any of claims 1-1 1 above wherein the said parameter or indicator relates to disease diagnosis or detection with or without reference to parameters as obtained by other more traditional techniques.

19. Apparatus to employ or harness the dynamic behaviour of specific temporal phases of erythrocyte and other blood related aggregation or agglutination m vitro or in vivo by using the dynamic behaviour of these temporal phases in the determination of at least one haematological parameter which may be a parameter with a numeric value or an indicator, said apparatus comprising a means of mixing for a predetermined time or with predefined shear and then to stop said mixing if In vitro or with means to halt or reduce flow similarly if in vivo and with means of selecting a defined region in the temporal evolution of moving aggregating erythrocytes or other such blood or blood related components undergoing aggregation or agglutination and comprising a means of making at least two time successive(non-simultaneous) remote examinations thereofto yield real or virtual measurement quantities which reflect the magnitude of which reflect the state or rate of or degree of completeness of the said aggregation and further comprising a means wherein the said quantities or their magnitudes are manipulated together mathematically so as to measure, or correlate with the said haematological parameter(s) or yield the said haematological indicator.

Apparatus as in claim 19 above wherein the means of remote examinations involves the supply and detection of shear or longitudinal (compression) wave energy of any frequency or waveform or any srhultaneous or sequential combination of frequencies and waveform. r1

21. Apparatus as in claim 19 wherein means of physical measurement may involve the supply and detection may be of transverse wave (electromagnetic) energy of any known polarsation and with a single constant exciting frequency when making the said two or measurements referred to 22 Apparatus as In claim 21 which involves the use of a separate single but different (preferably but not exclusively escalated) exciting frequency at each of the two or more said time successive measurements.

23.Apparatus as m any of claims 19, 21 and 22 wherein the said remote means of physical measurement may involve two or more time successive measurements of the electrical capacitance of the system as a whole, in accordance with a a prior art method and wherein preferably though not exclusively apparatus for capacitance measurement is as previously disclosed by the present inventor in priority document GB 2336685.

24.Apparatus as in any of claims 19-23 above with said parameter being haematocrit, measured preferably m the attractive phase as defined herein above.

25. ,Apparatus as m any of claims 19-23 with parameter as the mean cell volume measured preferably in the repulsive phase as defined herein above.

26. Apparatus as in any of claims 19-23 with the said parameter(s) relating to the haematocrit and/or cytoplasmic content and measured preferably m the quasi stable phase as deemed herem above. it

27. .Apparatus as In any of claims 19-23 above with the parameter being red cell count.

28. Apparatus as in any of claims 19-23 above with the said parameter or indicator relating to clotting function. 29. Apparatus as in any of claims 19-23 above with the said parameter or

indicator relating to blood type.

Apparatus as in any of claims 19-23 above with the said parameter or indicator relating to disease or infectious disease state diagnosis.

31. Apparatus as in any of claims 19 -23 above with the said parameter relating to the evaluation of drug delivery systems.

32. Apparatus as m claim 19 and any of claims 24 -31 wherein an energy source and energy loss probe are employed.

32 Apparatus as in claim 32 wherein said source and probe do not contact directly with the blood or blood related system 33. Apparatus as in claim 22 or 23 of claims herein above used in method as in claims or 11 above wherein said dynamic behaviour use of many successive said remote examinations at escalating frequencies accounts for observation of non relaxational, multiple relaxational or resonance type effects in an impedance scanning measurement.

A

34 Apparatus as In any of claims 19 -33 above used in vitro and which has energy source capable of penetrating patient sample I/D labels.

Apparatus as in any of claims 19 -33 above which has energy source capable of penetrating skin or nail.

Claims

Amended claims have been filed as follows

I.A method to employ or harness the dynamic behaviour of any of the specific early temporal phases of the aggregation of erythrocytes and /or blood components and blood related aggregations or agglutinations in vitro or in viva as discovered by the present inventor and as according to the present invention, which said dynamic behaviour, prior to sedimentation its state or rate or degree of allowing resultant determination of at least one haematological parameter (not sedimentation rate) which may be any other with a numeric value such as those of erythrocytes such as red cell count or mean cell volume or clotting function or an indicator such as blood type or the presence or absence of disease state comprising a first step if in vitro of mixing the blood to a predefined rate of shear or for a predetermined mixing time and then stopping the said mixing or if in viva halting or reducing its flow for a predetermined duration, comprising a second step selecting a defined region in the early temporal evolution of moving aggregating erythrocytes or other such blood or blood related components undergoing aggregation or agglutination prior to sedimentation end comprising athirdstep making at least two time successive (non -simultaneous) remote examinations thereof to yield real or virtual quantities which reflect the magnitude of which reflect the rate of or state of or degree of completeness of the said aggregation and further comprising a final step wherein the said quantities or their magnitudes are manipulated together mathematically so as to measure Determine or correlate with the said haematological parameter(s) or yield the said haematological indicator, either in viva or in vitro. L\
2. Method as in claim 1 where there is determination of two or more haematological parameters and wherein said remote examinations are preferably non-optical and involve the supply of energy from a source which can penetrate patient I/D labels or skin or nail and evaluation by a probe.
3..Method as in claim 2 wherein said probe measures energy loss.
4.Method as in claim 2 or 3 wherein said probe measures reflection.
5. Method as in any of claims 2-4 wherein said probe measures scattering
6. Method according to claim 2 above wherein the means of remote physical measurement may involve the supply and detection of shear or longitudinal (compression) -wave energy of any frequency or waveform or any simultaneous or sequential combination of frequencies and waveform.
7. Method according to the said claim 2 above wherein the said remote means of physical measurement involves the supply and detection of transverse wave electromagnetic energy of any known polarization and with a single constant exciting frequency when making the said two or measurements referred to herein above.
8. Method as in claims 2 and 7 above but involving the use of a separate single but different (preferably but not exclusively escalated) exciting frequency at each of the two or more said time successive measurements.
9 Method according to claim 1 above wherein the said remote examinations involve two or more time successive (sequential) measurements of the electrical capacitance ofthe system as a whole at fixed frequency, preferably within the range lkHz to 3GHz.
1 O.Method according to claims 1 and 9 wherein said time successive measurements involve measurement of electrical capacitance at different successive preferably escalating frequencies.
I 1.. Method as in either claim 9 or 10 wherein said capacitance measurement is in accordance with the prior art method for capacitance measurement as previously described by the present inventor in the disclosed priority document GB 2336685.
12. Method according to any of claims 1-11 above wherein above one of said parameters may be the haematocrit, measured preferably in the so called attractive phase of the aggregation as defined by the present inventor and disclosed by the present invention at preferably some 5-10 seconds abler mixing or halting of flow..
13. Method according to any of claims 1-12 above wherein one of said parameters may be the mean cell volume, measured preferably in the repulsive phase of the aggregation and where erythrocyte approach is limited as defined by the present inventor and disclosed by the present invention.
14. Method as in any of claims 1-13 above wherein one of said parameters relates to either the haematocrit or the cytoplasmic content, which is preferably measured in the quasi -stable phase of the aggregation as defined by the present inventor.
l 5. Method according to any of claims 1-14 wherein one of the said parameters is the red cell count.
16 Method according to any of claimsl-11 above wherein the said parameter or I indicator relates to clotting function.
17. Method as in any of claims 1-11 above wherein the said indicator relates to blood type.
18. Method as in any of claims 1-11 above wherein the said parameter or indicator relates to disease diagnosis or detection with or without reference to parameters as obtained by other more traditional techniques. am_
19. Apparatus to bring about the method described in claim 1 herein above in that it has the means to employ or harness the dynamic behaviour of specific temporal phases of erythrocyte and other blood related aggregation or agglutination in vitro or in viva by using the dynamic behaviour of these temporal phases in the determination of at least one haematological parameter, not sedimentation rate, which may be a parameter with a numeric value or an indicator, said apparatus comprising a means of mixing for a predetermined time or with predefined shear and then to stop said mixing if in vitro or with means to halt or reduce flow similarly if in vivo and with means of selecting a defined region in the temporal evolution of moving aggregating erythrocytes or other such blood or blood related components undergoing aggregation or agglutination prior to sedimentation and comprising a means of making at least two time successive(non-simultaneous) remote examinations thereofto yield real or virtual measurement quantities which reflect the magnitude of which reflect the state or rate of or degree of completeness of the said aggregation and further comprising a means wherein the said quantities or their magnitudes are manipulated together mathematically so as to measure, or correlate with the said haematological parameter(s) or yield the said haematological indicator.
20. Apparatus as in claim 19 above wherein the means of remote examinations involves the supply and detection of shear or longitudinal (compression) wave energy of any frequency or waveform or any simultaneous or sequential combination of frequencies and waveform.

AS
21. Apparatus as in claim 19 wherein means of physical measurement may involve the supply and detection may be of transverse wave (electromagnetic) energy of any known polarisation and with a single constant exciting frequency when making the said two or measurements referred to
22 Apparatus as in claim 21which involves the use of a separate single but different (preferably but not exclusively escalated) exciting frequency at each of the two or more said time successive measurements.
23.Apparatus as in any of claims 19, 21 and 22 wherein the said remote means of physical measurement may involve two or more time successive measurements of the electrical capacitance of the system as a whole, in accordance with a a prior art method and wherein preferably though not exclusively apparatus for capacitance measurement is as previously disclosed by the present inventor in priority document GB 2336685.
24.Apparatus as in any of claims 19-23 above with said parameter being haematocrit, measured preferably in the attractive phase as defined herein above.
25. Apparatus as in any of claims 19-23 with parameter as the mean cell volume measured preferably in the repulsive phase as defined herein above.
26. Apparatus as in any of claims 19-23 with the said parameter(s) relating to the haematocrit and/or cytoplasmic content and measured preferably in the quasi stable phase as defined herein above.
27. Apparatus as in any of claims 19-23 above with the parameter being red cell count.
28. Apparatus as in any of claims 19-23 above with the said parameter or indicator relating to clotting function.
29 Apparatus as in any of claims 19-23 above with the said parameter or indicator relating to blood type.
30. Apparatus as in any of claims 19-23 above with the said parameter or indicator relating to disease or infectious disease state diagnosis.
31. Apparatus as in any of claims 19 -23 above with the said parameter relating to the evaluation of drug delivery systems.

32. Apparatus as in claim 19 and any of claims 24 -31 wherein an energy source and energy loss probe are employed.
32. Apparatus as in claim 32 wherein said source and probe do not contact directly with the blood or blood related system.
33. Apparatus as in claim 22 or 23 of claims herein above used in method as in claims or 11 above wherein said dynamic behaviour use of many successive said remote examinations at escalating frequencies accounts for observation of non relaxational, multiple relaxational or resonance type effects in an impedance scanning measurement.
34. Apparatus as in any of claims 19 -33 above used in vitro and which has energy source capable of penetrating patient sample l/D labels.
35. Apparatus as in any of claims 19 -33 above which has energy source capable of penetrating skin or nail.