Towards Reliable and Quantitative Surface‐Enhanced Raman Scattering (SERS): From Key Parameters to Good Analytical Practice

Abstract Experimental results obtained in different laboratories world‐wide by researchers using surface‐enhanced Raman scattering (SERS) can differ significantly. We, an international team of scientists with long‐standing expertise in SERS, address this issue from our perspective by presenting considerations on reliable and quantitative SERS. The central idea of this joint effort is to highlight key parameters and pitfalls that are often encountered in the literature. To that end, we provide here a series of recommendations on: a) the characterization of solid and colloidal SERS substrates by correlative electron and optical microscopy and spectroscopy, b) on the determination of the SERS enhancement factor (EF), including suitable Raman reporter/probe molecules, and finally on c) good analytical practice. We hope that both newcomers and specialists will benefit from these recommendations to increase the inter‐laboratory comparability of experimental SERS results and further establish SERS as an analytical tool.


Introduction
Research in surface-enhanced Raman scattering (SERS) is ar apidly growing field. [1,2] It ranges from fundamental theoretical studies to the development of new enhancing materials/substrates for real-world applications.T hus,e ven for specialists,itisdifficult to maintain acoherent view of the entire field. In part this may be due to differences in the priorities of researchers who come from different backgrounds.Alarge number of variables which occur in atypical SERS measurement (see Figure 1) makes it very difficult to compare the data from different studies.H erein, we would like to make suggestions that we hope will allow researchers to have their work compared with other approaches,thereby highlighting the analytical advancements in the field. We would hope that this could also be used to give non-specialists an indication of which approaches might be most suitable for them and what performance they might expect to obtain. We begin by discussing ways in which the performance of SERS substrates can be reported, followed by highlighting the differences between solid and colloidal substrates (see Figure 2) as well as the challenge to thoroughly characterize nanoparticle suspensions.F inally,w e give recommendations of what would be useful to support further development of SERS-based chemical analysis in terms of good analytical practice.

Key Parameters in SERS and the SERS Enhancement Factor (EF)
As illustrated in Figure 1, at its simplest, the signal obtained in any SERS measurement is determined by four factors: i) Theidentity of the sample. ii)A number of molecules present in the enhanced region which is being probed.
Experimental results obtained in different laboratories world-wide by researchers using surface-enhanced Raman scattering (SERS) can differ significantly.W e, an international team of scientists with longstanding expertise in SERS,address this issue from our perspective by presenting considerations on reliable and quantitative SERS.The central idea of this joint effort is to highlight key parameters and pitfalls that are often encountered in the literature.T othat end, we provide here aseries of recommendations on:a )the characterization of solid and colloidal SERS substrates by correlative electron and optical microscopyand spectroscopy, b) on the determination of the SERS enhancement factor (EF), including suitable Raman reporter/ probe molecules,and finally on c) good analytical practice.W ehope that both newcomers and specialists will benefit from these recommendations to increase the inter-laboratory comparability of experimental SERS results and further establish SERS as an analytical tool. iii)T he amplification provided by the substrate. iv) Instrument settings and performance.
If we assume ak nown sample (i)w ith ad efined Raman scattering cross-section is being studied, this leaves just three factors which combine to give the observed signal. Although this apparently small number of factors seems relatively easy to compute and control, extracting acomplete set of figures of merit in aSERS measurement is extremely challenging.T his is partly because each of these factors is themselves determined by acombination of other variables and because some of the underlying variables are difficult to measure.
Forexample,inthe variable (ii)the number of molecules in the enhanced region will depend on both the area of asurface within the probed volume and the surface coverage. Additionally,itisimportant whether the analyte has ahigh or low affinity for the enhancing surface since this determines the residence time of the analyte within the probed volume.
Similarly,i n(iii)f or the enhancing SERS substrate there are numerous theoretical and experimental studies [1] that show the plasmonic enhancement provided by any substrate (typically Au,A g, Cu or Al) is governed by ac omplex interplay of the material used, the morphology of the structure,a nd the coupling of excitation photons and the surface plasmon. It has also been demonstrated that semiconductor substrates (GaP,S i) can provide large near-field enhancement and be used in surface-enhanced spectroscopies. [3] Finally,the instrumental factors in (iv) are acombination of the variables on the excitation side,s uch as the focusing conditions and the laser power, whereas on the detection side light-throughput, spectral resolution (grating/spectrometer), and detection efficiency( CCD/CMOS) influence how the generated SERS signal is dispersed and how much of it is actually detected.
Overall, the above considerations highlight why it is so challenging to directly and objectively compare SERS results obtained in different laboratories worldwide.T he most popular approach is to use an enhancement factor as am easure of the plasmonic enhancement and to define this in aw ay that aims to cancel out both the effects of instrumental factors and the intrinsic Raman scattering cross-sections of the test molecule.T his essentially provides afigure of merit of the effectiveness of the substrate and it can be transferred between laboratories.

Angewandte Chemie
Minireviews 5456 www.angewandte.org with I SERS ,t he SERS signal, I RS ,t he classical Raman signal obtained without the SERS substrate, N SERS ,t he number of molecules excited in SERS and N RS ,the number of molecules excited in classical Raman. TheEFcorresponds to the SERS intensity of one molecule divided by the Raman intensity of one molecule without the SERS substrate and can be seen as an absolute EF of the scattering cross-section of the test molecule.
There are some fundamental issues with this approach which are discussed below but even before that it is useful to note that accurate estimation of the EF requires that each parameter in the Equation (1) is measured accurately. I SERS is normally easy to measure as the SERS signal is normally large.T he estimation of the three other factors is more critical.
First, since the normal Raman scattering (RS) crosssections of molecules are very low,itisnecessary to make the I RS measurement on ah ighly concentrated solution or on ac rystal to be able to get as ufficient signal to noise (S/N) ratio.T hus,t he measurement cannot be done on molecules having the same state as the ones excited in SERS.I n addition, the Raman and the SERS spectra can also be different, as selection rules are modified due to specific interaction of the molecules with the substrate in SERS.Itis important to use the same vibrational mode to compare signals in RS and SERS,a sc ross-sections for different vibrations can differ greatly,a nd can experience different enhancement. This difference is mainly due to electronic interactions between the metal surface and the molecules. [5] It is also referred to as the "first layer effect" as it appears only for the molecules in direct contact with the metal. [6] In case of molecular electronic resonance (e.g., for dyes), it should be considered that the optical absorption cross-section of molecules can change when interacting with metallic nanoparticles because of modification of the intrinsic polarizability of the molecule upon adsorption and molecule-molecule interactions even at very low concentrations in the nm regime. [7] In such as ituation, the ignorance of (large) resonance shifts would lead to determination of an inaccurate SERS enhancement. To probe only the electromagnetic enhancement, it is possible to cover the metal surface with at hin dielectric layer (i.e.s ilica) as in the SHINERS approach. [8] However,o ther problems could arise,n otably when grafting molecules on the dielectric layer and because of the increased distance of the molecules from the nanoparticle surface.
Moreover,i ti sn ecessary to measure both I SERS and I RS under the same conditions,t hat is,t he same excitation wavelength and intensity,t he same focusing conditions, integration time and the same spectral resolution. Modern Raman instruments are nowadays often equipped with an EM-CCD and users should be aware that the amplification (EM gain) might lead to issues in terms of quantification, especially at low signal levels.
Second, the estimation of the number of molecules that take part in SERS and in normal Raman is not trivial. For I RS , it requires the excitation and collection volumes in the solution or in the solid sample to be known. With the concentration or the density of the sample,t he N RS can then be estimated. N SERS requires that the orientation and the surface coverage of the molecule be known. Assuming that the enhancement is localized on the metallic surface of the nanostructures, N SERS can be obtained from ak nowledge of the active surface area of the substrate that is being probed, the footprint of an adsorbed molecule (which can depend on orientation), and the surface coverage.E ven if 100 %c overage is assumed, N SERS can be difficult to determine accurately since the area can be difficult to define,o rt he interaction of the molecule with the surface may not take place as predicted. In the case of solid SERS substrates obtained by lithographic techniques,quantification of the surface area is facilitated by the periodic nature of the pattern, however, with colloidal nanostructures,the available area depends on the particle size and concentration. This means that the particle concentration used in aSERS experiment should be estimated and reported. Amorphous or disorganized rough noble metallic films obtained from metal sputtering or by adsorption of nanoparticles on surfaces are more challenging to characterize. Therefore,aconsideration of the uncertainty of EF estimate in such as ystem is even more important.
Am ore difficult problem is that, while the SERS EF in principle gives the plasmonic enhancement independent of the probe molecule,i np ractice the measured number will typically depend on the surface affinity of the probe molecule. Usually,the surface coverage is not determined by independent methods,but concentrations of analyte are kept high and complete surface coverage is assumed. However,i nr eality, the adsorption of the analyte onto the surface depends very strongly on the interaction between the analyte and the surface.Often, to reach the surface,the analyte must replace as urfactant or penetrate al ayer of as tabilizing species and consequently,t he SERS response I SERS depends strongly on the affinity of an analyte to the nanoparticle surface.T his means that for the same surface,s trongly and weakly adsorbing analytes at the same concentration will give different surface coverages,w ith the result that the weakly adsorbing analyte will give smaller SERS signals.T his can be misinterpreted as being due to different EF values,sothat the EF will depend on the nature of the test molecule.
Asolution could be the choice of a"neutral" analyte with potentially high binding affinity and the ability to displace the existing surface species or to co-adsorb.I nt he literature, several probe molecules having high affinity to silver or gold have been proposed, for example,a romatic thiols including thiophenol or 4-mercaptobenzoic acid, alkyl thiols (longer than C 5 but not longer than C 16 ), or molecules with an NH x group,s uch as adenine.S uch molecules have the advantages that they form an adsorbate on the metallic surface through S or Nb inding and they give characteristic SERS spectra that are easy to analyze.D ye molecules,s uch as crystal violet, methylene blue,orrhodamine 6G have also been used as they provide very high Raman cross-sections because of resonant enhancement, albeit with lower affinity to the surface through electrostatic interaction. Table 1l ists several potential SERS reporter/probe molecules that we consider to be useful, together with their respective advantages and disadvantages.W erecommend the strongly adsorbing,n on-resonant molecules 4-mercaptobenzoic acid (4-MBA, deprotonated at pH 10) and adenine as standard Raman reporters.Inthe future,the community will need to agree which less-strongly adsorbing compounds may also be suited to test accessibility of the surface of aS ERS substrate.W eemphasize that finding asingle strongly binding standard is an ecessary but not ad efinitive solution since it does not allow the accessibility of the surface to be measured. This is important because in most cases in which aparticular analyte gives no signal with areasonably plasmonically active substrate,itisbecause the analyte has not adsorbed onto the substrate rather than aproblem with the plasmonic enhancement. Poor binding may be caused by many different reasons, most obviously it may just be that the structure of the analyte means that surface binding is not thermodynamically favorable.This can be overcome by modifying the surface [9] but this is at opic not addressed herein. Alternatively,t he reason for low signals may be that the substrate is plasmonically active, but the surface is passivated by as urface layer, for example protein which prevents access by all but the most strongly binding analytes.S uch substrates will show large EFs with strongly binding test molecules but not be useful in practice since they will not allow detection of more weakly bound species.W e, therefore,p ropose that in addition to very strongly binding test molecules,new substrates are also tested with weaker binding molecules so that the accessibility of the surface may be characterized. If this is done,t hen both the plasmonic efficiencyand the ability to detect weaker binding molecules will be known and any compromises associated with ag iven substrate will be made apparent. Forw eakly adsorbing analytes,s olid SERS substrates or ligand-free nanoparticles (e.g.,p roduced by laser ablation in liquids) might be preferable since they do not contain acapping agent on the surface.For strongly adsorbing analytes,SERS colloids with acapping agent are suitable as well.
Another way to estimate the SERS signal enhancement is to measure ar elative EF.I nt his case,areference sample is used to estimate as tandard intensity (I standard )t hat is reproducible in any conditions and any environment. For simplicity and to be easy to obtain, such standard sample should be as olid sample as aS is urface with specific orientation and specific polarized excitation, as imple polymer such as polystyrene or an organic solvent. [4] It can also be an internal standard inserted with the probe molecule as inherently included molecules on the SERS substrate [10] or an isotopically labelled version of the molecular probe with aknown relative concentration [4,11] TheS ERS intensity I SERS could then be compared to this I standard and the EF variation or optimization would be estimated by comparison with this I standard .T his approach has the advantage of allowing the comparison of I SERS measured with different instrumental conditions and equipment and also avoids the need to estimate the values of variables that are used to calculate the N SERS and N RS .Such amethodology could be especially useful for inter-laboratory and intersample comparisons.F or inter-sample comparisons,t he comparison should also be done using the same probe molecules (e.g.4 -MBA, adenine), as the enhancement can include acontribution due to the chemical effect.
Considering all these parameters,d rawing fair and quantitative comparisons between various SERS experiments is challenging.However,ifall the details of the experimental conditions and the assumptions used in the calculation are given, especially those concerning the state of the molecules on the SERS substrate and on the reference sample used to measure the I RS and N RS ,t here is no fundamental problem with using this approach to estimate the plasmonic efficiency.

Types of SERS Substrates and Their Characterization
SERS metallic substrates can be classified in three basic categories ( Figure 2): [12] 1) nanostructures fabricated directly on asolid substrate by lithography and template techniques in at op-down approach [13] 2) nanoparticles assembled and/or immobilized on solid substrates in abottom-up approach, and 3) nanoparticles in suspension (bare,J anus or core-shell of different compositions including silica-, polymer-coated, and shell-isolated nanoparticles (SHINs) [8] metal-encapsulated dielectric core nanoparticles,b imetallic and core-satellite configurations). Representatives from the three categories exhibit significant differences with respect to the homogeneity of their geometrical structure,i nstrumentation, and know-how required for fabrication/synthesis,a nd the option to be scaled up.I na ddition to common metallic substrates, some semiconductors [14] and 2D inorganic materials [15] can provide reliable and quantitative SERS measurement.
Thespectral reproducibility and the sensitivity of any new solid SERS substrate should be compared with commercially available substrates (Silmeco,H amamatsu, ST Japan etc.)o r standard home-made substrates,such as nanoarrays prepared by nanosphere or colloidal lithography,m etal film over nanospheres (MFON), or thin metal film (4 nm) on glass. [16] Also new colloidal SERS substrates should be compared with commercially available substrates (BBI, nanoComposix, Nanopartz, NanoWerke etc.) or standard home-made 20-50 nm gold/silver colloids prepared by the Turkevich (Au, citrate) or Leopold/Lendl (Ag, hydroxylamine) method. [17] Although silver is generally plasmonically more active than gold, oxidation is aproblem. In the case of home-made solid and colloidal SERS substrates,detailed preparation protocols should be provided.
TheS ERS substrate plays ac rucial role as the electromagnetic enhancement is related to the material, size,a nd shape of the nanostructure. [1,18] Currently,considerable effort continues to be devoted to the preparation of novel and ever more sophisticated SERS substrates.W hile theoretical analysis and modelling of their electromagnetic properties can underpin their rational design and fabrication it is also critical that standardized methods are used in their characterization. In particular, electron microscopy and extinction/reflectance spectroscopy can provide an idea of the shape,s ize distribution, and plasmon resonance for metallic nanostructures.F or substrates on as olid support, scanning electron microscopy (SEM) and scanning probe microscopies,such as atomic force microscopy (AFM), scanning tunneling microscopy (STM), and scanning near-field optical microcopy (SNOM) can provide valuable information on the structures created. However,w hile such measurements are often the only ones that are reported for novel substrates,t his is typically not sufficient to predict their SERS performance.T he central problem is the dramatic spatial variations in local optical fields within af ew nanometers that are predicted by theory and observed in experiments which make the enhancement hugely dependent on very small structural features.These can be studied by other methods,asdiscussed below but it means that simple imaging of as ubstrate is not ag ood guide to the SERS enhancement that it may produce and it should not be used for this purpose.C onversely,l ocal optical fields can be probed by excitation of the plasmon with an electron beam in the highly regular arrangements on nanostructures fabricated on solid supports and at the single-aggregate level with great precision by excitation of the plasmon with an electron beam. Electron energy loss spectroscopy (EELS) and optically excited local field intensities can differ, so both EELS and optical excitation could be used for plasmonic characterization. By EELS,s hifts of plasmon resonances with small changes in gap size can be mapped. [19] We recommend combining several characterization techniques in acorrelative approach on the same sample.T his also facilitates the comparison with predictions from computer simulations.
Forc olloidal SERS substrates,t ransmission electron microscopy (TEM) can be used to obtain information on nanoparticle size distribution and shapes,p rovided that ap roper statistical evaluation is performed. Dynamic light scattering (DLS) can be particularly useful for nanoparticles of narrow size distribution, and for an assessment of nanoaggregate formation when analyte molecules are added. The potential interaction with analyte molecules can be studied by determining the zeta potential. Although usually provided by the SERS spectrum itself,X -ray photoelectron spectroscopy (XPS) and electrochemical approaches can be used to evaluate the interactions at the interface between the plasmonic nanostructure and the molecules to greater detail. Importantly,t he interaction with light can be simulated for most of the cases,giving insight into,for example,the spectral response,e lectric-field confinement, and polarization-dependence.H owever,t he information provided by these characterization techniques depends on the system under study and should be judged critically.T he first question that we need to keep in mind-and try to answer using these methods and techniques-is:where is the signal in our SERS measurements coming from?Itisgenerally accepted that the SERS signal predominantly arises from molecules located in very small (few nm 3 or even less [20] )r egions with extremely high local electric field, so-called hot spots.Asimple calculation of an elongated Ag nanoparticle shows that 12 %o ft he total surface area is responsible for 80 %o ft he collected SERS signal. These numbers can be even more striking when considering Ag sphere dimers with ag ap of 2nm(see Figure 3, right). In that case,only 0.59 %ofthe total surface contributes to 80 %ofthe measured SERS signal (see Figure 3, left). [21] Fort his reason, it is important to keep in mind that what we are seeing,s ensing,t argeting,s tudying in SERS is usually at iny fraction of the total adsorbed molecules. [22] Theh ighly localized nature of the hot spots forces us to rethink the validity of the standard characterization methods for colloids or nanostructured substrates when trying to link them to their SERS performance.T his strong dependence on nanoscale inhomogeneities makes it challenging to predict enhancement factors or reproduce results obtained by different groups using similar colloids or substrates.Our characterization efforts should be centered on accessing to these regions/molecules to fairly reproduce results and correlate them with theory/simulations.
Overall, we strongly recommend that the plasmonic characterization must be carried out at the level of individual nanoparticles and nanoaggregates by correlative techniques based on ac ombination of electron microscopy,d ark-field and SERS microspectroscopy on the same sample. [23] Generally,t he optical characterization should be performed before electron microscopy owing to potential sample damage caused by electron microscopy.C ommercial providers offer,f or example,i nstrumentation for correlative Raman/ SEM (for example,Raman:WITec, SEM:T escan and Zeiss).

Good Analytical Practice
In this Section, we point out to some of the analytical shortfalls that in our opinion should not be further perpetuated. In SERS-based quantification of analytes,acalibration model of the dependence of aS ERS signal on the concentration of an analyte is typically used to predict the concentration of this analyte in an unknown sample.W es trongly recommend that in addition to the calibration procedure also av alidation step is performed to make sure the SERS detection can do more than making accurate predictions on the set of data that it was calibrated with. [24] In other words: aS ERS-based detection scheme without av alidation step should be questioned regarding its relevance to an analytical application.
Most SERS studies devoted to demonstrating analyte quantification capability tend to focus on the ultra-sensitivity of SERS through the lens of the limit of detection (LOD) and sometimes limit of quantification, namely the lowest concentrations that the sensor is capable of detecting and quantifying, respectively.H owever,m ore important than the LOD is the ability to predict concentrations with accuracy in the range of concentrations relevant to the actual concentration likely to be encountered in the targeted samples.T herefore, sensor development should aim for the concentration range that matches the specific quantification problem at play,f or example,as pecific physiological concentration in bio-detection.
It is useful to note that the LOD is often used as ameans of characterizing the performance of novel SERS substrates. While this is valid if the objective is to demonstrate detection of as pecific target analyte,i tis more often used as as imple general indication of SERS performance.T he obvious problem with this approach is that the LOD depends not only on the substrate used and the sample concentration but also on instrumental factors,s uch as the collection efficiencyo rt he grating efficiencya nd detector sensitivity.T hus,h igh-performance instruments will give higher signal to noise ratios than lower ones with the same sample.T his means that LOD comparisons should really be limited to side-by-side compar- Figure 3. Calculated enhancement factor (EF) distribution around the hot spot in adimer of silver spheres (radius 30 nm, gap 2nm). The excitationw avelength is taken at the dipolar localized surface plasmon resonance, which provides the maximum SERS EF. The logarithmic false color maps show graphically the surface SERS EF distribution in the j E j 4 approximation. The plot on the left shows the SERS EF in the plane of incidence as afunction of arc length L along the surface together with the maximum SERS EF (F max ), the surface-averaged SERS EF (hFi)a nd the relative area a 80 from which 80 %ofthe total SERS signal originates.Figure adapted from Ref. [21].
isons of substrates.F or inter-laboratory comparisons the instrument performance should also be reported, for example by showing the normal Raman spectrum of astandard solvent along with clearly stated experimental conditions.
In addition to the sensitivity range,two other indicators of performance that are routine in the evaluation of chemical analysis methods should be reported. First, the recovery rate, which is the ratio of the detected concentration to the actual concentration in the sample;t his allows estimating whether the sensor is overestimating or underestimating the actual analyte concentration. Second, the root mean square error (RMSE) of prediction, which gives an estimate of the precision of the SERS sensorsr eadout, should be determined. [25] Form aximal relevance to the application, this last figure of merit is best calculated on the validation data set to test the predictive power of the sensor on samples which are known to the experimenter but unknown to the calibration process.
Asignificant issue with quantitative SERS measurements is the reproducibility.F or colloids this is typically equated with the extent to which different preparations of nominally the same particles give the same absolute SERS intensity;for solids this should be linked with signals given by different production batches,b ut it is often confused with uniformity, that is,the signal given at different points on the same surface. It is useful to have an indication of variation both within and between batches and we would recommend in case of colloidal nanoparticles in suspension to average the results of three different batches and at least 10 individual measurements per batch under the same conditions (when long integration times of ca. 10 sare needed) and ideally more than 100 individual measurements (for short integration times around 1s). In the case of assemblies or other solid substrates, depending on the complexity of the fabrication and uniformity of the materials,m easurements of tens of points on asurface can give an approximate indication of the uniformity of as ubstrate;t oo btain statistically significant results SERS data should be averaged from 3-5 samples with afew 100 sof individual measurements per sample recorded under the same conditions (typically short integration times < 1s)a nd distributed over the reported area. [26] It is important to note that there are significant limits to the application of SERS to complex real-world samples in which there may be issues with interference from the nontarget constituents.O ften, switching to "real-world" samples from "clean-laboratory" samples introduces the problem of interference from impurities in the sample matrix. As examples,blocking of the SERS substrate surface or changed plasmonic properties of the substrate,orstrong SERS signals from other compounds should be mentioned. This illustrates the importance of realistic sample matrices also in the laboratory setting.S uch problems may be reduced by pretreating the sample to remove interfering species [27] or by chemically modifying the surface of the SERS substrate,f or example,w ith self-assembled monolayers,o re ven with species that promote binding of the target species. [9,28] Such approaches improve selectivity at the cost of the general applicability of aSERS sensor.
As discussed above,changes in the EF of the substrate in the presence of analyte species must be properly addressed, specifically in quantification. Of course,h aving reasonably reproducible substrates is helpful, as is using as ampling method designed to minimize variation. Fore xample,t he incubation of SERS substrate in asolution containing analyte yields SERS signal with al ower variability than the droplet deposition. [29] However,t hese precautions may still not be sufficient to prevent variations owing to the sample matrix effects.
Fortunately,E Fv ariations can be mitigated by the addition of as tandard whose response will change in the same way as the target to experimental variations.The target signal can then be normalized to the internal standard signal, see the Review article Ref. [30].T he most obvious internal standards are isotopologues of the target since these will track the variation in the EF of the target very well. [31] Alternatively,chemically similar compounds may be appropriate [32] or standard addition can be used. [33] Afurther approach is the rational design and synthesis of SERS nanotags (typically core-shell nanoparticles) where the standard (SERS-active label) can be embedded directly between core and shell such as in core-molecule-shell (CMS) nanoparticles. [34] An additional advantage of the introduction of internal standards is that it dramatically reduces the need for rigorous batch-tobatch reproducibility of substrates and spectral acquisition. We,t herefore,w ould like to emphasize that by using an appropriate internal intensity standard quantitative SERS analysis can be achieved even without very highly reproducible substrates.
Finally,w ew ould like to point out that the presented attempt on formulating recommendations for the practice of SERS is certainly just astarting point for future discussions in the community and that we are looking forward to feedback from our colleagues.