DE102006045618A1 - Measuring arrangement for quantitative analytics, has micro-channel section with transparent detection window and self-regenerating thin film that passes separation fluid into micro-channel and section between segments and channel wall - Google Patents

Abstract

The arrangement has a resonance-Raman-spectrometer (1), and a micro-channel (4) filled with a fluid and connected with a device for discharging liquids. A micro-channel section (31) is provided with an optically transparent detection window (5), and forms a flow cell (2) and an outlet opening (6) of the arrangement. The flow cell is adapted and adjusted to a detection position, where the cell is made of silicon and glass. A self-regenerating thin film passes a separation fluid into another micro-channel (3) and the micro-channel section between segments and a channel wall. An independent claim is also included for a method for quantitative analytics by using the nano-particle-reinforced Raman- and/or resonance Raman spectroscopy using the measuring arrangement.

Description

The The invention relates to a measuring arrangement and a measuring method for the quantitative Analysis based on nanoparticle-enhanced Raman spectroscopy or resonance Raman spectroscopy based and integrating the preagregation of nanoparticles under defined, long-term stable process conditions in the detection process enable.

The Infrared spectroscopy and Raman spectroscopy are finding wide Application in structure elucidation and for the proof of identity organic compounds.

This is how the Scripture reveals DE 10 2004 034 354 B3 For example, an ultra-compact infrared Raman spectrometer (handheld device), which should be used for investigations of chemical structures, etc. or for the quasi-simultaneous identification of many bioparticles on substrates, the location of the sample to be examined directly in the Positioning the optical spectral unit is positioned, the optical excitation of the sample to be examined by means of focused radiation of the laser exactly in the plane of the virtual gap position of the imaging spectral unit and in the beam path between the sample and spectral unit exclusively a Raman radiation transmitting filter is arranged.

conditioned due to the high information density of the vibrational spectra of organic matter molecules and the pronounced individual character of the spectra of a compound are these Method of vibrational spectroscopy also on the quantification of analytes in complex analytical matrices.

advantage of Raman spectroscopy infrared absorption spectroscopy apart from a clear band assignment, above all the fact that Raman spectra in aqueous Medium can be measured since water has only weak Raman-active bands and thus it is not comes to an overlay.

a Another advantage is the use of the technique of resonance-enhanced Raman Spectroscopy. Using this technique, desired structure groups succeed selectively stimulate and the vibrational spectra of the excited structural elements capture. It will be for Resonantly excited vibration modes amplifications by up to 9 orders of magnitude observed which is a reliable Separation of the desired Ensuring signals from the background of the analytical matrix.

For the excitation of the resonance electronic transitions in the analyte molecule are used. By choosing different excitation wavelengths, certain structural units in a molecule can be selectively excited. The efficiency of the resonance enhancement correlates with the extinction of the absorption band of the respective structural unit (see contributions to the Special Issue: J. Raman Spectrosc. 2001, 32, 399 .)

A further increase in signal intensity can be exploited by the use of local field enhancements, as can be achieved by plasmonic effects in the immediate vicinity of nanoparticles and aggregated nanoparticles (nanoparticle clusters). In this case, nanoparticle aggregates prove to be particularly efficient. Methods for their preparation from stabilized nanoparticle suspensions are described in the literature ( Faulds, K .; Littleford, RE; Graham, D .; Dent, G .; Smith, WE, Comparison of Surface-Enhanced Resonance Raman Scattering from Unaggregated and Aggregated Nanoparticles. Analytical Chemistry 2004, 76, (3), 592-598 ). In this case, further signal amplification is achieved for analyte molecules which are in close proximity to the nanoparticles. As an additional selection criterion, the adsorption of the analyte molecule on the surface of the nanoparticle cluster can be used in this case. As a result, nanoparticle-enhanced resonance Raman spectroscopy enables the direct detection of analytes down to the detection of single molecules ( Kneipp, K., Raman spectroscopy of single molecules. Single Molecules 2001, 2, (4), 291-292 ).

In addition, surface-enhanced resonance Raman spectroscopy has the potential for highly specific, parallel, label-free quantification of multiple analyte molecules in complex analytical matrices ( Docherty, FT; Monaghan, PB; Keir, R .; Graham, D .; Smith, WE; Cooper, JM, The first SERRS multiplexed from labeled oligonucleotides in a microfluidics lab-on-a-chip. Chemical Communications (Cambridge, United Kingdom) 2004, (1), 118-119 ).

The disadvantage of this analysis method, however, is that it due to the high energy density of the excitation light in the detection cuvette for photodeposition of nanoparticle / analyte conjugates to the Küvet tenfensten comes. When using microchannels, the adhesion of NP-analyte conjugates causes the occurrence of a so-called "memory effect". This problem has hitherto prevented the use of this method as a spectrally resolved detection technique for flow injection analysis (FIA).

One obvious approach to circumvent this effect is the cleaning the cuvette window with a suitable cleaning solution before and after each analysis step. However, this approach limits the sample rate and requires the use of aggressive chemicals and hazardous substances for this step.

Another disadvantage of the known analysis method is the large temporal fluctuation of the SERS signal after addition of additional Aggregierunsfluid. ( Schneider, S .; Gray, H .; Halbig, P .; Nickel, U., Kinetic effects in surface-enhanced Raman spectroscopy: does it have any potential as an analytical tool? Analyst (Cambridge, United Kingdom) 1993, 118, (6), 689-94 ) and the rapid aging of the pre-aggregated nanoparticle suspension. This must be generated immediately before the detection and mixed with the analyte. In the course of aging, there is a rapid change in the achievable with this solution resonance amplification.

Also known is the generation of fluid segments that are transported in microchannel systems, as well as the metered addition of fluids to these fluid segments [see T. Henkel, T. Bermig, M. Kielpinski, A. Grodrian, J. Metze, JM Köhler: Chip modules for generation and manipulation of fluid segments for micro serial flow processes. Chemical Engineering Journal 101 (2004) pp. 430-445 such as B. Zheng, JD Tice, and RF Ismagilov: Formation of Droplets of Alternating Composition in Microfluidic Channels and Applications to Indexing Concentrations in Droplet-Based Assays. Anal. Chem. 76 (2004) pp. 4977-4982 ].

From the publication Galder Cristobal, Laurence Arbouet, Flavie Sarrazin, David Talaga, Jean-Luc Bruneel, Mathieu Joanicot, and Laurent Servant: On-line laser Raman spectroscopic probing of droplets engineered into microfluidic devices, Lab Chip, 2006, 6, 1140-1146 the use of classical Raman spectroscopy for the analysis of droplet sequences generated in microfluidic devices is known.

Of the Disadvantage of this analysis method is that they are in their sensitivity is limited because there is no signal amplification by nanoparticles. Would be at this known method, a gain by nanoparticles for Use, would this lead to an adhesion of these on the cuvette window. These NP / analyte conjugates would be only by cleaning the cuvette window with a suitable cleaning solution before and after the analysis step of each individual sequence drop to eliminate what is practically unrealizable.

The The object of the invention is that mentioned above To avoid disadvantages of the prior art, in particular a Specify measuring arrangement and a measuring method for quantitative analysis, those on the nanoparticles reinforced Raman, as well as resonance Raman spectroscopy are based and integration the preagregation of the nanoparticles under defined process conditions enable in the detection process.

According to the invention This task is characterized by the characterizing features of the first and the 22nd claim and supplemented by advantageous embodiments according to the subclaims.

The Essence of the invention is that the use of liquid / liquid two-phase flows in microchannels for the Generation of nanoparticle-analyte segments separated by the separation medium is used, the metered addition of aggregation fluid to these Nanoparticle analyte segments is optionally possible, so that below detection by surface-enhanced Raman or resonance Raman spectroscopy can be done.

In contrast to the prior art, the measuring arrangement according to the invention and the inventive method allow the quantification of organic molecules in a concentration range between 0.1 and 1 .mu.mol / l. This corresponds to 10 ⁹ molecules / segment.

Essential to the invention is that the formation of a self-regenerating thin film of separation fluid in the capillary gap between the segments and the channel wall for the suppression of photodeposition of nanoparticle aggregates is used in a microfluidic arrangement. This effect is supported by the special flow conditions in liquid / liquid segmented sample streams and the specific flow regime of the Taylor flow encountered here. High velocity gradients are observed in the embedded segments in near-wall areas [see A. Günther, M. Jhunjhunwala, M. Thalmann, MA Schmidt, KF Jensen: Micromixing of Miscible Liquids in Segmented Gas / liquid flow. Langmuir 21 (2005) pp. 1547-1555 such as D. Malsch, M. Kielpinski, R. Merthan, J. Albert, G. Mayer, JM Köhler, H. Süße, M. Stahl, T. Henkel: μPIV Analysis of Taylor Flow in Microchannels, Accepted for presentation to International Conferece to Micro Reaction Technology IMRET 9, Potsdam 6.9.2006-8.9.2006 , submitted for publication in Chemical Engineering Journal], which additionally counteract the attachment of nanoparticle clusters.

According to the invention this per se known effect in the measuring arrangement according to the invention and in the inventive method for the efficient suppression of photodeposition of nanoparticle aggregates used at the detection windows.

The Invention will be described below with reference to the schematic drawing of embodiments explained in more detail. there demonstrate:

1 FIG. 2: an embodiment of the measuring arrangement according to the invention in a schematic overview,

2a : a section from 1 in a sectional view and

2 B : a section from 2a ,

In the 1 The measuring arrangement shown comprises a Raman spectrometer 1 and a flow cell adapted and adjusted to its detection position 2 , wherein this flow cell 2 with a device 7 for conveying separation fluid fluidly via a first microchannel 3 is connected and with this liquid-filled micro-channel 3 one or more with devices 7 for the promotion of liquids associated further micro-channels 4 are fluidically connected.

This first microchannel 3 flows as in 2a shown in a with at least one optically transparent detection window 5 provided micro-channel section 31 , which at the same time the outlet opening 6 the Messanordung forms.

The cross section of the first microchannel 3 is preferably in the range between 0.3 mm ² and 0.001 mm ² .

The cross section of the first microchannel 3 opening further microchannels 4 range is between 0.3 mm ² and 0.001 mm ² .

The cross-section of the mouth of the opening microchannels 4 can be equipped with an additional, narrow edged narrowing. In this case, the cross section of the junction in the first microchannel 3 opening microchannels 4 about 0.3 mm ² to 0.001 mm ² .

Particularly advantageous is the optically transparent detection window 5 planar. Alternatively, however, it may also be curved.

The detection window 5 consists of a crystalline or amorphous, transparent inorganic material, for example quartz, quartz glass or glass.

Alternatively, the detection window 5 from a composite of inorganic or organic materials having thermoplastic or thermosetting properties, for example inorganic or organic polymers.

The microchannels 3 ; 4 have advantageously rounded edges and may, for example, have a circular cross-section.

Alternatively, the microchannels 3 ; 4 but also have a quadrangular cross-section.

The flow cell 2 advantageously consists of a composite of silicon and glass, in particular a composite of several glass semi-finished products.

A self-regenerating thin film is advantageous 9 of separation fluid in the microchannels 3 ; 4 and the flow cell 2 , which due to the functionalization of the channel wall between the Segments and the channel wall forms ( 2 B ). Functionalization means that at least the first microchannel 3 and the microchannel section 31 are provided with a layer which improves the wetting of the channel walls by the separation liquid.

there For example, the layer may be a molecular film, for example of alkyl or aryl silanes and their condensation products, wherein the alkyl or aryl silanes in addition Amino, carboxy, hydroxy or sulfonic acid groups may have.

alternative For this purpose, the layer of polymers, such as. Polyacrylates, polyester or polyolefins, these additionally being amino, carboxy, hydroxy or sulfonic acid groups can own.

It is particularly advantageous if the microchannels 3 ; 4 and the flow cell 2 coated with organic polymers that improve the wetting of the channel walls and the cuvette walls by the separation medium, resulting in a thin boundary layer between the channel wall and segments.

alternative this coating can be obtained by treatment with chlorosilanes, alkoxysilanes, Acyloxysilane or hexamethydisilazane be formed.

alternative For this purpose, the wetting of the channel wall by the separation medium be improved by means of plasma treatment.

In the method using the measuring arrangement according to the invention are in a continuously traversed by a separation fluid microchannel 3 Test fluid-containing segments produced from the fluids NP suspension and analyte, which subsequently by the as a flow cell 2 executed microanalysis section 31 are passed and registered as the passage of the segments through the detection area surface-enhanced Raman or resonance Raman spectra of the test fluid contained in the segment.

alternative can do this the segments of the test fluid from the fluids NP suspension, aggregation fluid and analyte are generated.

• 1. Erzeugung von Segmenten enthaltend Nanopartikel-Suspension in einem Trägerstrom vom Separations-Fluid im ersten Mikrokanal 3,
• 2. Zudosierung von Analyt zu den Segmenten im ersten Mikrokanal 3 durch Dosieroperationen oder Fusionierung vermittels eines weiteren Mikrokanals 4, wobei zuvor eine Durchmischung und Inkubation in einer Verweilschleife 8 erfolgen kann,
• 3. Zudosieren von Aggregations-Fluid zu den Segmenten im ersten Mikrokanal 3 durch Dosieroperationen oder Fusionierung mit Aggreations-Fluid enthaltenden Segmenten vermittels eines Mikrokanals 4, wobei zuvor eine Durchmischung und Inkubation in einer Verweilschleife 8 erfolgen kann, und
• 4. Vermessung des erzeugten Test-Fluids im Detektionsbereich durch das Detektionsfenster 5 hindurch vermittels des Resonanz-Raman Spektrometers 1,

wobei die zeitliche Abfolge der Prozessschritte 1 und 2 wahlfrei ist und der Prozessschritt 3 optional enthalten sein kann, falls ein zusätzlicher Aggregationsschritt benötigt wird.The method according to the invention comprises the following steps:

• 1. Generation of segments containing nanoparticle suspension in a carrier stream from the separation fluid in the first microchannel 3 .
• 2. Addition of analyte to the segments in the first microchannel 3 by metering operations or fusing by means of another microchannel 4 , where previously a mixing and incubation in a residence loop 8th can be done
• 3. Metering of aggregation fluid to the segments in the first microchannel 3 by metering operations or fusion with segments containing aggreation fluid by means of a microchannel 4 , where previously a mixing and incubation in a residence loop 8th can be done, and
• 4. Measurement of the generated test fluid in the detection area through the detection window 5 through the resonance Raman spectrometer 1 .

the timing of the process steps 1 and 2 is optional and the process step 3 may optionally be included if an additional aggregation step is needed.

The advantage of the measuring arrangement according to the invention and of the measuring method according to the invention is that in comparison with the prior art Galder Cristobal, Laurence Arbouet, Flavie Sarrazin, David Talaga, Jean-Luc Bruneel, Mathieu Joanicot, and Laurent Servant: On-line laser Raman spectroscopic probing of droplets engineered into microfluidic devices, Lab Chip, 2006, 6, 1140-1146 , a 10 ⁷ to 10 ⁸ times higher measuring sensitivity in continuous flow analysis, which is in the range of low-throughput efficient single cuvette detection.

The Measuring arrangement according to the invention and the method according to the invention will be explained in more detail below with reference to the embodiment, without that limited to this become.

embodiment

In the measuring arrangement according to the invention is a Raman microscope Labram Fa. Horiba-Jobin-Yvon and a microsystem-produced flow meter (microchannel chip), containing egg NEN main channel and T-shaped opening secondary channels used. For this purpose, in the first embodiment, a flow measuring cell with a size of 16 × 25 mm and equipped with a main channel, two opening channels, a dwelling and a detection area is used.

In the second embodiment is the flow cell with a total of five opening channels and one cuvette area fitted.

When Separation fluid is used tetradecane. Analyte, nanoparticle suspension and aggregation fluid are applied as an aqueous solution or suspension.

1st embodiment

The Microchannel chips are used as a composite of two glass wafers made of glass, borofloat 33, Schott, Jena, diameter 100 mm, thickness 0.7 mm and manufactured subsequently by sawing sporadically.

Everyone This glass wafer is with half channels in a mirror image arrangement fitted.

The half channels are assembled by anodic bonding so that full channels arise.

The microchannels be by etching of the glass with hydrofluoric acid using a 100 nm thick nickel / chromium coating as an etching mask produced.

The main channels are separated by a width of the mask opening of 330 microns Channels through a width of the mask opening of 30 μm Are defined. The mask openings the merging Channels end 200 μm before the mask opening of the main channel.

On These are the result of etching at the junctions Nozzle structures, which the process reliability of the generation of segments in the Positively affect dosing operations.

The etching depth of the channels is 135 μm. For the Connection of HPLC capillaries be at the ends of the microchannels Connecting holes with a diameter of 0.5 mm generated by ultrasonic luffing.

Subsequently, will on one of the substrates on the channel side, a 60 nm thick auxiliary bonding layer, consisting of polycrystalline silicon deposited by sputtering. The so prepared half channels are adjusted to each other and with the method of anodic Bonded together.

in the The result will be microchannel chips with the dimensions (width × height) of 600 × 270 μm for the main channel and the cuvette and (width × height) 300 × 270 μm for the opening channels receive.

To Removal of the polysilicon layer from the microchannels Treatment with a 5% solution of sodium hydroxide in water at 50 ° C and rinsing with water are suitable obtained optically transparent microchannel chips, which in the field of the detection window on top and bottom by planar allele Be limited observation window with a width of 330 microns.

In front the use of the flow cell, the inner walls of the microchannels be chemically modified so that they, for, the separation fluid used here Have tetradecane wetting properties.

This is by treatment of the channels with a solution of octadecyltrichlorosilane (2 mmol / L) in anhydrous n-heptane at 50 ° C over a Period of 2 hours.

Excess reagent is by rinsing the microchannels washed with anhydrous n-heptane. Alternatively, the Wetting properties by coating with a polymer film be set. Therefor In the first step, the treatment of the microchannels with hexamethyldisilazane takes place (10 min), followed by dry-blowing of the channels with air and heating 120 ° C over a Period of 20 min.

On this, serving as an adhesion-promoting layer molecular film of polydimethylsilane takes place the deposition of a polymer film by filling the microchannels with a solution of perfluoroalkülgruppen-bearing polymer (Teflon AF, DuPont) in perfluorooctane and subsequent blowing excess Polymer solution with air and drying at 120 ° C over a period of 1 h.

The Fluidic connection of the flow measuring cell takes place with the help of of standard HPLC devices attached to the fluid port holes be pressed on. The promotion the fluid is made using a 4-channel syringe pump system from Cetoni using 500 μl precision syringes. By the Main channel is the separation fluid tetradecane at a flow rate of 0.1 μl / s promoted.

At the first one Channel is the generation of segments containing nanoparticles by advancement of the nanoparticle suspension in this channel at a flow rate of 0.02 μl / s.

At the second confluence the metered addition of a mixture of analyte and aggregation fluid.

2nd embodiment

at the second embodiment be the same process of manufacturing and coating the channel walls as in the first embodiment applied.

Of the Main channel becomes semicircular over the Chip arranged and with five opening out Provided channels.

The Separation fluid tetradecane is delivered at a flow rate of 0.1 μl / s.

at this design becomes the first opening channel for the generation used by water-containing segments at a flow rate of 0.02 μl / s, into which in the second opening Channel nanoparticle solution with flow rates between 0.005 and 0.02 μl / s.

On this way you can Segments are generated with variable nanoparticle concentration.

At the third confluence the metered addition of aggregation fluid with variable flow rates between 0.005 and 0.002 μl / s and at the 4th fatigue the metered addition of the analyte solution with freely selectable, variable flow rates in the range between 0.005 and 0.02 μl / s. The remaining, fifth junction can for The metered addition of another optional component is optional become.

When using the embodiment 1, the following results are obtained:
The colloid used is a citrate-reduced partially pre-aggregated gold colloid. The analyte used is the dye crystal violet. The flow rates are adjusted to 0.02 μl / s for analyte and nanoparticle suspension and 0.1 μl / s for the separation medium tetradecane and kept constant throughout the measurements.

Raman spectra are recorded continuously with an integration time of 1 s per spectrum, with the laser focus in the channel in x-, y- and z-direction while the measurement is kept constant to reflect signal fluctuations an inhomogeneous particle distribution within the aqueous To avoid dripping.

The 1 illustrates the applied measuring principle.

Spectrum A shows the Raman spectrum of the separation medium tetradecane measured in the oil phase. In contrast, in spectrum B, which is measured in an aqueous segment, the band structure of the SERS spectrum of crystal violet can be seen. The labeled band at a wave number value of 1177 cm ^-1 , which is used in the following to calculate the integrated Raman intensity, is due to an in plane deformation mode of the ring. The left panel of the graph shows the integrated Raman intensity in a wavenumber range of 1130-1217 cm ^-1 as a function of the measurement time. One can recognize a regularly alternating pattern of a crystal violet containing aqueous segment and the separation medium tetradecane, which has no bands in the region used to calculate the Raman integrated intensity. Due to a signal ratio of I (B) / I (A) = 600, a clear delimitation of the crystal violet-containing aqueous phase from the separation medium tetradecane is possible.

If the flow of crystal violet is stopped during a series of measurements, crystal violet can no longer be detected in the aqueous segments. (See 2 )

In the aqueous Segment, which now consists of pure colloid, can now neither a SERS spectrum of crystal violet nor a Raman spectrum of the tetradecane be measured. (see A). This proves the fact that the aqueous droplet from an oil film which is an adhesion of nanoparticle aggregates to the cuvette prevented.

To perform a quantitative analysis, crystal violet solutions at various concentrations of 10 ^-5 to 10 ^-6 M are measured. In this case, a measurement series is recorded for each concentration over a period of about 15 minutes. During this period about 80 crystal violet segments can be measured. For the quantitative evaluation, the mean value is then determined via the peak peaks of the integrated Raman intensity and this is then plotted against the respective crystal violet concentration.

This results in a linear dependence of the calculated mean of the crystal violet concentration (see 3 ).

The regression coefficient of the linear regression is R ² = 0.9823. All features shown in the description, the following embodiment, the claims and drawings can be essential to the invention both individually and in any combination.

Table 1

Definition of terms:

• separation fluid

  denotes a preferred Raman-inactive fluid

  analyte

  denotes a solution of sample to be analyzed, containing the components to be analyzed and beyond other components.

  NP suspension

  denotes a long-term stable Suspension of nanoparticles in a fluid

  Aggregation fluid

  denotes a fluid, which contains reagents that cause aggregation of the nanoparticles.

  ANP suspension

  denotes a suspension aggregated nanoparticles

  Test fluid

  refers to an ANP suspension and analyte-containing fluid associated with the separation fluid is immiscible.

  Raman

  inactive As Raman inactive substances are called which under the selected detection conditions and detection areas no significant contribution to the Raman vibrational spectrum or their vibrational bands clearly from the bands of Analyte molecules can be separated.

Miscibility matrix Table 2

• 1:

  miscible

  0:

  immiscible

Separations fluid analyte NP suspension Aggregation fluid ANP suspension Test fluid Separations fluid 1 0 0 0 0 0 Analyte fluid 1 1 1 1 1 NP suspension 1 1 1 1 Aggregation fluid 1 1 1 ANP suspension 1 1 Test fluid 1

Wetting matrix Table 3

• 1:

  wetting

  0:

  not wetting

channel wall Separations fluid 1 analyte 0 NP suspension 0 Aggregation fluid 0 ANP suspension 0 Test fluid 0

1

Resonance Raman spectrometer

11

laser

12

spectral detector

2

Flow Cell

3

first microchannel

31

Microchannel portion

4

Another microchannel

5

detection window

6

outlet

7

contraption to promote of separation fluid

8th

Verweilschleife

9

thin film

Claims (25)

Measuring arrangement for quantitative analysis by means of nanoparticle-enhanced Raman spectroscopy or resonance Raman spectroscopy comprising • a resonance Raman spectrometer ( 1 ), • a flow cuvette adapted and adjusted to its detection position ( 2 ), whereby this flow cuvette ( 2 ) with a device for conveying separation fluid fluidly via a first microchannel ( 3 ) and filled with separation liquid, and at least one further microchannel filled with a liquid ( 4 ) with a device for conveying fluids, which fluidly with the first microchannel ( 3 ), the first microchannel ( 3 ) into a with at least one optically transparent detection window ( 5 ) provided microchannel section ( 31 ), the flow cuvette ( 2 ) and at the same time the outlet opening ( 6 ) forms the Messanordung, and wherein a self-regenerating thin film ( 9 ) of separation fluid at least in the first microchannel ( 3 ) and the microchannel section ( 31 ) between the segments and the channel wall. Measuring arrangement according to claim 1, characterized in that at least the first microchannel ( 3 ) and the microchannel section ( 31 ) are provided with a layer which improves the wetting of the channel walls by the separation liquid. Measuring arrangement according to claim 2, characterized in that the layer is a molecular film is. Measuring arrangement according to claim 3, characterized in that the molecular film of alkyl or aryl-silanes consists. Measuring arrangement according to claim 4, characterized in that the alkyl or aryl silanes additionally amino, Carboxy, hydroxy or sulfonic acid have. Measuring arrangement according to claim 2, characterized in that the layer consists of polymers. Measuring arrangement according to claim 6, characterized in that the polymers polyacrylates, polyesters or polyolefins. Measuring arrangement according to claim 7, characterized in that the polyacrylates, polyesters or polyolefins additionally Have amino, carboxy, hydroxy or sulfonic acid groups. Measuring arrangement according to claim 2, characterized in that the layer by chlorosilanes, Alkoxysilanes, acyloxysilanes or hexamethydisilazane is generated. Measuring arrangement according to claim 1, characterized in that the cross section of the first microchannel ( 3 ) Is 0.3 mm ² and 0.001 mm ² . Measuring arrangement according to claim 1, characterized in that the cross section of the first microchannel ( 3 ) opening microchannel ( 4 ) Is 0.3 mm ² and 0.001 mm ² . Measuring arrangement according to claim 1, characterized in that the cross-section of the mouth of the opening microchannel ( 4 ) is equipped with an additional, narrow-edged narrowing. Measuring arrangement according to claim 11, characterized in that the cross-section of the mouth of the first microchannel ( 3 ) opening microchannel ( 4 ) Is 0.3 mm ² to 0.001 mm ² . Measuring arrangement according to claim 1, characterized in that the optically transparent detection window ( 5 ) is planar. Measuring arrangement according to claim 1, characterized in that the optically transparent detection window ( 5 ) is curved. Measuring arrangement according to claim 1, characterized in that the detection window ( 5 ) consists of a crystalline or amorphous, transparent inorganic material. Measuring arrangement according to claim 1, characterized in that the detection window ( 5 ) consists of a composite of inorganic or organic materials with thermoplastic or thermosetting properties. Measuring arrangement according to claim 1, characterized in that the microchannels ( 3 ; 4 ) have rounded edges. Measuring arrangement according to claim 1, characterized in that the microchannels ( 3 ; 4 ) have a circular cross-section. Measuring arrangement according to claim 1, characterized in that the microchannels ( 3 ; 4 ) has a quadrangular cross-section. Measuring arrangement according to claim 1, characterized in that the flow cell ( 2 ) consists of a composite of silicon and glass. Procedure for the quantitative analysis by means of the nanoparticle-reinforced Raman or resonance Raman spectroscopy using the measuring arrangement according to one or more of the preceding claims in which one continuous micro-channel test fluid through which a separation fluid flows containing segments are generated from the fluids NP suspension and analyte, which subsequently by the designed as a flow cell channel section be passed and the passage of the segments through the detection area surface-enhanced Raman or resonance Raman spectra of the test fluid contained in the segment are registered. A method according to claim 22 comprising the following steps: a) Generation of segments containing nanoparticle suspension in a carrier stream from the separation fluid in the first microchannel 3 . b) Addition of analyte to the segments in the first microchannel 3 by metering operations or fusing by means of another microchannel 4 , where previously a mixing and incubation in a residence loop 8th and c) measuring the generated test fluid in the detection area through the detection window 5 through the resonance Raman spectrometer 1 , A method according to claim 22, characterized in that between step b) and c) the following step is in between: • metering of aggregation fluid to the segments in the first microchannel 3 by metering operations or fusion with segments containing aggreation fluid by means of a microchannel 4 , where previously a mixing and incubation in a residence loop 8th can be done. Procedure for the quantitative analysis by means of the nanoparticle-reinforced Raman Spectroscopy using the measuring arrangement according to a or more of the preceding claims in which the segments of Test fluids from the fluids NP suspension, aggregation fluid and Analyte be generated.