COLLOIDS AND SURFACES Colloids and Surtees ELSEVIER A Physicochemical and Engineering Aspects 107 (1996) 87-75 Review Applications of atomic-force microscopy in environmental colloid and surface chemistry Patricia A. Maurice Dept. of Geology, Kent State University, Kent, OH 44242, USA Received 10 March 1995; accepted 30 June 1995 Abstract Atomic-force microscopy (AFM) is a powerful technique for imaging mineral surfaces in air or immersed in solution and at sub-nanometer-scale resolution. As the technique continues to develop, AFM is emerging as an important means not only for imaging the surface structure and microtopography of environmental particles, but also for determining changes in microtopography over the course of dissolution, growth, sorption, heterogeneous nucleation, and redox reactions. Additionally, AFM can provide a direct means of probing the structure of the double layer. As AFM use continues to proliferate, there is an increasing need for critical evaluation of image acquisition and processing techniques. This manuscript reviews the “state of the art” in force microscopy with regard to theoretical considerations, technological advances, and current and potential applications. Some common artifacts are discussed, with the purpose both of preventing new users from re-making old mistakes and of enabling non-users to evaluate AFM results. Finally, examples of applications in colloid and surface chemistry are used to illustrate the potential of AFM for providing new insight into a wide range of environmental processes. Keywords: Atomic-force microscopy; Clay; Fulvie acid; Hematite; Microbial dissolution; Scanning-probe microscopy; Surface Table of contents 1. Introduction 2. Basic operating principles 3. Tip-sample interactions 3.1. Forces between the tip and the sample 3.2. Tapping-mode AFM 3.3. Tip shape considerations 4. Examples of applications 4.1. Atomic-scale imaging: importance of forces 4.2. AFM as a probe of double-layer forces 43. Inesitu imaging of surface microtopography 44. Imaging particulates (0927-7757)96/$15.00.© 1996 Elsevier Science B.V. All rights reserved SSDI 0927-7757(95)03372-68 PA, Maurice/Collos Surfaces A: Physicochem. Eng, Aspects 107 (1996) $7 75 4.5. Clay particle morphology and thickness measurements 4.6. TMAFM imaging of humic substances 5. Conclusions Acknowledgments References 1, Introduction ‘Atomic-force microscopy (AFM; also known as scanning-force microscopy, SFM) was developed in 1985 [1], and it quickly captured the attention of the environmental chemistry community as a long-awaited technique for imaging mineral sur- faces in air or immersed in solution, and at sub- nanometer-scale resolution. As a result, applica- tions by environmental geochemists and soil chem- ists began to appear in the literature by the early 1990's [2-5]. The past few years have seen a rapid proliferation of AFM use by members of the environmental chemistry community, particularly as more user friendly and versatile commercial instruments are made available. Applications within the realm of environmental surface chemis try include determining the molecular- to atomic- scale structure of mineral surfaces [6], probing forces at the mineral-water interface [7-9], mea- suring directly the kinetics of growth, dissolution, heterogeneous nucleation and redox processes [10-17]. elucidating the effects of varying sorption mechanisms over the course of electrochemical reactions [18], visualizing sorption of mactomo- lecular organic substances and hemimicelles [19], imaging soil aggregates [20], and determining the thicknesses and morphology of clay-sized particles (21-2417), As a relatively new technique, the theory and applications of AFM are still in the development stage. As more laboratories join the AFM com- munity, widespread critical review is needed. The purpose of this paper is to review the “state of the art” in force microscopy as applied to research in environmental colloid and surface chemistry. Basic operating principles will be reviewed, followed by a discussion of tip-sample interactions. Some common artifacts will be discussed, with the pur- pose both of preventing new users from re-making old mistakes and of enabling non-users to evaluate AFM results. Finally, examples of applications in environmental colloid and surface chemistry will be presented. The overall purpose of this paper is not to provide an exhaustive review, but rather to give the reader a sense of the ever-increasing capabilities of AFM, while at the same time point- ing out some of the potential problems that need to be addressed. 2. Basie operating principles Because the author of this article works primar- ily with commercial atomic-force microscopes pro- duced by Digital Instruments, Inc. (Nanoscope 11 and Nanoscope IIT), the discussion that follows will be biased toward the Digital Instruments machines. Previously unpublished results of the author's research presented here were collected using Nanoscope II and II AFMs. Examples from the literature, however, draw on research con- ducted with many different commercial and custom-built machines. The basic operating principles of AFM have been described in detail many times in the literature [1,3]. A brief review is provided here to set the stage for a discussion of image collection and interpretation. Briefly, AFM works by rastering a sample mounted on a piezoelectric tube under a sharp tip that is attached to, or part of, a cantilever. Deflection of the cantilever as the sample is moved thereunder can be monitored by a variety of means Most commonly, it is monitored via an optical ever consisting of laser light reflected off the end of the cantilever toward a photodiode. AFM imaging may be conducted in one of two modes. In the constant-force mode, a feedback loop is used to adjust the height of the sample to keep the cantilever deflection constant during sample raster- ing. In the so-called constant-height mode, the response of the z-piezo is more sluggish so thatPA. Maurice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) 57-75 9 the height of the sample remains more nearly constant, and the cantilever changes its deflection as the sample is scanned thereunder. The constant- height mode is not appropriate for surfaces with a large amount of topographic relief, because the sample surface may be damaged as the z-piezo fails to compensate for large changes in relief, such that the tip drags against relatively large topo- graphic features. As shown in Fig. 1, the essential elements of APM are the tip, the cantilever, the detector (laser/ cantilever/photodiode system), the scanner, the computer control system and, of course, the sample. Each of these elements can contribute its, own set of limitations and artifacts in the imaging process (see Fig.2). Because the interaction between the tip and the sample is the most crucial element for AFM imaging, it exerts the greatest control on the types of samples that can be imaged, and under which imaging conditions, and contrib- utes the greatest number of potential artifacts and interferences. In the discussion that follows, we shall focus on the tip, the sample, and tip-sample interactions. More comprehensive reviews are pro- vided by Sarid [25] and Eggleston [26]. Laser Photodetector 2 Piezoelectric ‘Scanner Fig 1, Schematic illustration of the atomic-force microscope, showing that the essential elements are the tip, the cantilever, the detector (laser/cantilever/photodiode system), the scanner, the computer control, and the sample. Each of these elements, ‘may contribute its own set of limitations and artifacts ‘Some Potential Limitations, Problems, and Artifacts in Atomic Force Microscopy ‘Other components and | | external effects eelimgamme ||| Someerconts | See ||] Reker | Piezoelectric scanner rate lt unas | ||| Steno ern | Saas = || SESE || [_Fnteannne {2h delat unos weed | | eiotmie merce Esnonst cane Ermer ||] enenen Een oe Fig. 2, Some potential limitations, problems, and common arlifacts associated with the various elements of atomic force ‘microscopy. This paper focuses on the tip, the sample, and tip sample interactions; for more extensive discussion see Refs (25] and [26] 3, Tip-sample interactions 3.1. Forces between the tip and the sample Although a detailed understanding of the forces between the tip and the sample in air and aqueous media is crucial for image collection and interpreta- tion, these forces are still poorly understood and the focus of some controversy. At the root of this problem lies our incomplete understanding of the forces at interfaces and between surfaces, in general. Indeed, it can be said that AFM and the science of interfacial forces are evolving synergistically, as theories of interfacial forces are used to explain imaging phenomena [27-33] while force spectro- scopy is used to probe near-surface forces [7-9]. ‘The discussion that follows will focus on contact- mode AFM, which is the most commonly used imaging mode. Non-contact-mode AFM and tap- ping-mode AFM are also important techniques and they will be discussed later in the text.0 PA, Maurice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) 57-75 Both attractive (F,) and repulsive (F,) forces ‘may contribute to the total forces (F,) between the tip and the sample (see Fig. 3). The most important attractive forces appear to be van der Waals (vdW) interaction and capillary adhesion forces. The vdW forces are long range and relatively weak. Their origin is quantum mechanical in nature. They result from a variety of interactions, primarily induced dipole and quadrupole interactions. Israelachvili [34] provides an excellent explana- tion of the induced dipole interactions. In short, even a non-polar atom will experience an in- stantaneous dipole moment due to instantaneous asymmetries of the positions of protons and electrons. This will generate an electric field which, in turn, will polarize nearby atoms, resulting in an instantaneous attractive force between the atoms [34]. The vdW forces are non-localized; ie. they are spread out over many atoms, For force micro- scopy, Goodman and Garcia [30] estimated vdW forces in the order of 10-20 nN. They also esti- mated that for metallic tips and samples, vdW forces may be as high as about 100 nN when the AFM is operating in the purely repulsive mode [30] Capillary adhesion is an important attractive force during imaging in air, This capillary force results from the formation of a meniscus made up of water and organic contaminants sorbed on the surface of the tip and the sample [9] (see Fig. 3). The capillary force is large and has been estimated to be in the order of 100 nN or more [35]. When the tip and the sample are completely immersed in [eee water or another liquid, a meniscus does not form and hence capillary forces are absent. The primary repulsive force at close approach is responsible for the original name of the instrument, the atomic-force microscope. This repulsive force occurs between any two atoms or molecules at close approach, when their electron orbitals begin to overlap [34]. It is thus a result of the so-called Pauli exclusion principle. Prediction of the magni- tude of this force is complicated by unknown factors such as the amount of surface-atomic relax- ation [27,30,31,36]. Repulsive forces also may arise from solvation or hydration forces. The water neat hydrophilic surfaces is structured [34]. When the tip and the sample are brought into close contact during force microscopy, resistance, and hence apparent repulsion, arises as the structured water molecules on the surfaces of the tip and the sample are pushed away. Contact-mode AFM is generally operated with a net repulsive force of between 10 and 100 nN; however, greater forces are possible and need to be minimized. Eppell et al. [37] reported hard-core repulsive forces of approxi- mately 100 nN, while imaging in air proteins deposited on mica. Finally, in aqueous solutions, electrical double- layer forces, which may be either attractive or repulsive, may be present near the surfaces of the tip and the sample, These double-layer forces arise because surfaces in aqueous solution are generally charged. The surface charge may come about in three principal ways: (1) permanent structural charge due to defects or substitutions in the crystal TRA + h) Fig 3. Schematic illustration showing the forces present during AFM imaging in air, The (otal force, Ps the sum of the attractive van der Waals and capillary forces, — F,, and the repulsive force, F,, between the tip and the nple, Note the presence of a meniscus, which increases the attractive forces between the tip and the sample, When the tip and sample are immersed in solution, this capillary force is not present, and both the attractive force and the total force between the tip and the sample are decreased,PA, Maurice/Collods Surfaces A: Physicochem. Eng. Aspects 107 (1996) 37-75 6 lattice [38]; (2) hydroxylation and ionization of surface functional groups (formation of >SOH groups) [39,40]; (3) adsorption of ions from solu- tion, which may either create charge on a pre- viously uncharged surface or alter the direction and magnitude of charge. Considering the contributions from (1) and (2), the intrinsic surface charge density (¢,,) on the tip ‘or sample material is equal to the sum of the permanent structural surface charge density (¢o) and the net proton surface charge density (oy) which results from hydroxylation [38] Gin= 00+ Ou i) Focusing on oy, if hydroxylation results in unequal amounts of H” and OH” on the surface, then the density of surface charge contributed by hydroxyla- tion and ionization, 04, will be given by on=ely—Ton) (2) where € is the absolute value of the charge on one electron and I’ refers to a surface excess concen- tration. At some pH characteristic of a given surface material, [fy] = on. This value of pH is known as the point of zero net proton condition (PHpanpe)> If we neglect the importance of other contribu- tions to surface charge, such as ions adsorbed from solution, then the point of zero charge (pHyac; here used as a relatively inexact term) of a given material is related to its chemical composition [40]. Hence, we can expect that different tip and sample materials will have different pH. values, and will be positively charged, negatively charged, or uncharged over different ranges of pH. Thus, the electrostatic force between tip and sample will be sensitive to pH and may change from attractive to repulsive (or vice versa) if the pH changes. In aqueous solutions containing electrolytes, this simple scheme is complicated by the fact that constituents other than H* and OH™ may adsorb specifically onto surfaces, forming complexes that can also contribute (o the surface charge density and hence to the electrostatic forces between tip and sample. In addition to specifically sorbed constituents, positively charged ions will accumulate in solution near negatively charged surfaces and negatively charged ions will accumulate near positively charged surfaces, in order to balance the charge on the surface. This accumulation of counterions will be greatest close to the surface and will decrease with distance away from the surface. The accumulation of ions near surfaces forms what is known as the electric double layer (EDL); the thickness of the double layer (defined as the Debye length, 1/x) depends on a number of parameters. Pethaps most importantly, it is inversely related to the solution ionic strength (I), according to the equation (for symmetrically charged electrolytes, eg. Nat and Cl} =(2F?T x 10°ee9RT)"? @) where F is the Faraday constant, ¢ is the dielectric constant of water, € is the free space permittivity, R is the molar gas constant, and T is the absolute temperature (K) [41]. When AFM is conducted in aqueous solution, two surfaces, “the tip and the sample”, are brought into close proximity, and their EDLs interact. This interaction may be either attractive or repulsive and of different magnitude, depending on the sur- face-charge properties of the tip and sample materi- als, and on solution parameters including the types and concentrations of ionic constituents, ionic strength, pH, and temperature. If we know the PHpzc of the tip and sample materials, and the concentrations of ions in solution, then we may attempt to predict the effects of double-layer forces in AFM. However, because theories of double- layer interactions generally deal with flat surfaces or spherical colloidal particles [34], applications involving sharp tips may require further refine- ments, The effects of double-layer forces on AFM images are only now beginning to be explored (see below) [7-9]. Experimentation using tip and sample materials with different surface-charge properties represents a growing field in force microscopy. An important step in understanding the forces between the tip and the sample is to examine the force-versus-distance curve that results as the tip is brought into contact with the sample. Fig. 4, adapted from Weisenhorn et al. [9], shows a typical force-versus-distance curve in air. The x- axis in Fig. 4 represents the vertical movement ofe P.A, Maurice/Collotds Surfaces A: Physicochem. Eng, Aspects 107 (1996) 57-75 Ropulsive touching Distance Fig 4. A typical force-versus-distance curve for contact-mode AFM, adapted from Weisenhorn ct al. [9]. See text for a description of the components of the curve the piezoelectric translator. Moving left along this axis corresponds to the tip coming closer to the sample surface; 0 represents “just touching". The y-axis represents the photodiode (detector) signal, which is proportional to the cantilever deflection and hence to the force between the tip and the sample. Above 0, the tip is being repelled, or deflected upwards. Below 0, the tip is being attracted or pulled downward toward the surface (9) At the far right of the plot, the tip is far from the sample surface. As the tip is brought closer to the surface (moving left on the plot), a point is eventually reached at which the tip contacts the surface. The forces on initial contact are complex and not completely understood, but certainly include attractive vdW forces as well as capillary forces as described above. If the sample is moved yet closer to the tip, the force increases along the diagonal line, as shown. If the sample is moved downward, withdrawing the tip from the sample, there is hysteresis in the curve due to attractive forces between the tip and the sample. The force decreases along the diagonal line below the zero- force line. Eventually, a point is reached at which the force is minimal, and the tip detaches. The force at the detachment point is known as the pull- out force [9]. Most AFM imaging is performed in the repulsive regime (upper left of Fig. 4), with a net repulsive force of between 10 and 100 nN, and is known as contact-mode imaging. Attractive- regime imaging also can be conducted and may be preferable for atomic-scale applications in solution, as described below [31,32]. Surfaces in air contain a small amount of sorbed water, and organic and other contaminants. When samples are imaged by AFM in air a menis- cus forms between the tip and the sample (Fig. 3). This meniscus results in a relatively strong attrac~ tive capillary or adhesion force of the order of 100 nN [9], in addition to the attractive vdW force typically of the order of 10's of nN and repulsive tip/sample contact forces of 10-100 nN. Thus, Drake et al. [42] estimate a relatively large force betwee the tip and the sample in air, typically greater than 100 nN. When the tip and the sample are completely immersed in aqueous solution, this meniscus does not form, and forces can be much lower. Weisenhorn et al. [35] report forces as low as 2 nN, and AFM can be controlled to give forces of the order of 10°! N or less [31]. Because the sample surface is rastered beneath the tip, frictional lateral forces must also be taken into account. At low forces, a linear relation should hold between the lateral force, F,, and the force normal (vertical) to the surface, F,. with a propor- tionality constant equal to the coefficient of friction, 4 [26]. Warmack et al. [43] showed this linear relation to be valid to a repulsive force of about 30 nN. It has been shown that frictional forces can influence the contrast of the AFM image when operating in the contact (repulsive) mode [44]. Frictional forces have been shown to vary on an atomic scale, and with temperature, scan velocity, relative humidity, and tip and sample material [45.46]. Overney et al. [46] showed that by simultaneously measuring both the normal and lateral (frictional) forces on the scanning tip, different compositional domains on Langmuir: Blodgett films could be determined. The resolution of the instrument was 10-"° N in the frictional force mode. The modified AFM which allows these measurements is known as the frictional force microscope (FFM). Frictional forces, adhesive forces, and chemical interactions between tip and sample can result in etching of a sample surface along step edges and other features, during the scanning process. Delawski and Parkinson [45] showed that the edges of pits were eroded parallel to the scan direction. They attributed this erosion to higher frictional forces as the tip was “trapped” in thePA. Maurice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) $7-7S 6 etch pit. The erosion rate was found to correlate linearly with force. Cratering, or the build-up of material along the edges of the etched areas, was observed only at applied forces greater than 150 nN. Multiple images were common during erosive imaging, probably owing to build-up of eroded ‘material on the tip surface (see Section 3.3). Further discussion on erosion of the sample surface is given in Section 4.3. A technique for avoiding damage of the sample surface is so-called non-contact-mode AFM. In this technique, the tip is kept a small distance above the sample surface and is given a small oscillation; weak attractive vdW forces are detected, Non-contact-mode AFM can result in reduced resolution relative to contact-mode AFM. ‘Also, conducting non-contact-mode AFM in solu- tion can be difficult, because the vaW forces tend to be weaker [47] 3.2. Tapping-mode AFM Tapping-mode® AFM (TMAFM) [48] is a new technique patented by Digital Instruments that circumvents the problem of large lateral frictional forces by applying an alternating current to the tip-cantilever system, causing the tip to vibrate and to contact the sample surface numerous times for each data point. TMAFM is excellent for the imaging of soft and easily deformable environmen- tal particles. It has proven especially useful for imaging particles deposited on other surfaces (see below). Although TMAFM could not originally be used in solution, a new form of solution TMAFM has been developed. The author has not yet had an opportunity to test this new technique. 3.3. Tip shape considerations Most AFM imaging in environmental chemistry has been conducted with commercially available silicon nitride (SijN,) integral cantilever-tips These tips are pyramidal in shape and have a radius of curvature 2500 A when new. With use, the tips are abraded resulting in an increased radius of curvature, For relatively small steps and other features with low relief, the radius of curva- ture at the end of a typical SisN, tip can limit the resolution of the step and particle images, making the features appear too broad [49]. Wilson et al [50] demonstrated that AFM images of biomolec~ ules and other structures with heights in the order of 10's of nanometers often are enlarged by as much as 25% owing to the finite size of the tip. For large-scale features, tip shape can convalve with surface features, often producing misleading results. In such cases, the sides of the tip contact the edges of features before the apex of the tip comes into contact. The solid angle (pyramidal) of a standard SixN, tip is about 55°, and the steepest vertical features that can be accurately imaged are thus about 625° [24]. The identification of tip- related artifacts can often be difficult; hence, the experience of the microscopist becomes invaluable. Excellent reviews such as that by Griffith and Grigg [51] are helping to make microscopists aware of problems, limitations, and potential solu- tions. Once a particular artifact such as a reverse- tip image (see below) is properly identified, the microscopist is alerted to the fact that similar structures imaged on other samples may also be artifacts. In the extreme case of very sharp features, such as whisker crystals, on sample surfaces, a reverse image of the tip may result [52]. The process controlling such images is illustrated in Fig. 5(a); a reverse-tip image is shown in Fig. 5(b). These images show structures related to the tip shape, primarily pyramidal features with triangular facets. Images of the type shown in Fig.5(b) may be helpful to the experimentalist in that they are pronounced examples of what may be more subtle effects along step or particle edges, or wherever a sample surface feature is steeper than the tip struc- ture. This problem can be particularly frustrating in the imaging of sub-micron to micron-scale particles, as shown in Fig. 5(c). Comparison with Fig. 5(b) suggests that the hematite particles in Fig. 5(¢) are probably more representative of the tip shape than of the particle shape, because trian- gular facets and pyramidal features occur which are similar in scale and structure to the features seen in the reverse-tip image. More subtle effects also can occur, such as sloping particle walls Kepler and Gewirth [53] noted that different types of tips will result in different shapes of artifacts;64 P.A, Mawrice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) 57-75 Fig 5. (a) Schematic illustration of how scanning a sharp feature can produce a reverse-ip image. For case of illustration the tip position is depicted as moving horizontally, whereas in reality the sample is scanned under the tip. Tip imaging is a ‘common artifact (se, for example, Ref.[52), particularly on rough surfaces. (b) AFM image in air of a sharp feature on a surface, producing a reverse image of the tip, as the edges of the tip contact the surface feature. (c) AFM image of hematite particles on pe membrane filter; tip-sample convolution results, in featuees similae those in (b), for example, they found square pyramidal tip, sample convolution artifacts with a Digital In- struments Nanoscope tip-cantilever, but triangu- lar artifacts with a Park Scientific Ultralever tip- ver. Finally, frictional forces between tip and sample may result in frictional tip/sample convolu- tion artifacts; such artifacts generally have an L-shaped outline as the tip drags across a sharp surface feature. Tip dimensions also can limit the ability of the tip to image narrow, deep features such as steep- sided pits. In this case, the pit will appear to be v-shaped, as the edges of the tip meet the sides of the pit before the tip reaches the bottom; in con- trast, if the pit depth exceeds the imaging height parameters, the pit may appear to be too shallow and flat-bottomed (off-scale), In attempts to circumvent the problems posed by tip shape limitations, tips have been developed with high aspect ratios or smaller radii of curvature [54,55]. These new tip designs are limited by the fact that sharper tips tend to become fragile. Thus, a number of researchers have proposed methods and algorithms for improving images using decon- volution procedures. As pointed out by Griffith and Grigg [51], the term deconvolution is not strictly correct because the tip and sample interact in a non-linear fashion; nevertheless, the term will be used here for simplicity. Some researchers have relied on purely mathematical treatments in which the geometry is assumed to be an idealized shape, such as spherical. However, more precise methods are possible. Markiewicz and Goh [56] back- calculated the tip shape by imaging samples (poly- bead-amino microspheres) of known size and geometry, recognizing that the AFM image will be a convolution of tip and sample shape. Calculated tip shapes were then used to deconvolute addi- tional samples. Wilson et al. [50] used 14 nm gold spheres in a similar procedure, and found good agreement between back-calculated or “restored” tip shapes and actual SEM images of tips. Deconvolution procedures such as these need to be calibrated and standardized, and necessary algo- rithms incorporated into AFM software. Irregular tip shapes can pose additional prob- lems. On the atomic scale, multiple (more than single-atom) tips can lead to a confusing array of apparent atomic-scale surface structures. ‘The effects of multiple-atom tips on imaging at the atomic scale have been the subject of numerousPA, Maurce/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) $7-7 65 investigations (see, for example, Ref. [27}), which should continue in light of the recent re-evaluation of atomic-scale imaging by Ohnesorge and Binnig [31]. At topographic scales, tips with irregular structures can lead to equally erroneous and poten- tially misleading results. For example, in Fig. 6, there are numerous twin rounded particles on the surface whose regularity in structure and orienta- tion suggests that they resulted from a double tip. Such effects can be particularly pronounced and confusing if the tip shape changes repeatedly over the course of an imaging session. Figs. 7(a)-7(c) show an example encountered during an experi- ment wherein a single crystal of specular hematite ‘was immersed in citric acid, with the purpose of imaging changes in microtopography over the course of dissolution. All of the images were col- lected with the scan direction from right to lef. ‘The sequence begins with a large island on the left-hand side of the image, which appears to have a step along its right edge. As imaging continues, two more steps appear to the right of this edge, and then all the steps disappear. Continued imaging resulted in steps appearing and disappear- ing repeatedly. When the scan direction was reversed at the end of the sequence (not shown Fig. 6. AFM image in air of a muscovite (001) surface following, reaction with deionized water, Numerous pairs of rounded particles are observed, whose regularity in structure and orientation suggests that they result from a double-tip configu ration. This is a common artifact, especially when imaging surfaces containing particulates that may adhere to the tip or imaging surfaces that arc easily eroded, here), a step appeared along the left edge of the island while the step on the right edge disappeared. All of the supposed steps are, in fact, clear examples of artifacts due to changing tip shape among various single and multiple configurations. 4, Examples of applications 4.1. Atomic-scale imaging: importance of forces Since the advent of AFM, the question of whether or not it can provide true atomic-scale resolution has remained controversial. This contro- versy revolves around the fact that most published atomic-scale images show perfectly ordered pe odic arrays, with none of the smail-scale defects such as kink sites commonly present in STM images [57]; in addition, the heights of atomic corrugations are often inordinately large [49]. In 1993, Gerhard Binnig, one of the inventors of AFM, published a landmark paper which is helping to resolve this controversy. In a study of calcite structure, Ohnesorge and Binnig [31] demon- strated that true atomic-scale resolution, including, imaging of monoatomic steps, could be achieved only when the estimated net repulsive loading force was <10"! N. Given that the typical force con- stant for a chemical bond is in the order of 100 Nm”, tip-sample interaction forces greater than 100 nN, as commonly encountered in repulsive- mode imaging in air, would displace the atoms in a bond by nanometer-scale distances [26]. Such strong forces would probably break bonds and damage the sample surface. At such high forces, the tip may be driven into the sample such that the contact point between the tip and sample would consist of several atoms. Images with period- icity related to crystallographic structure can occur at higher forces, but the images do not show individual atoms, and high forces can distort rela- tive atomic positions. Imaging in solution, and in the attractive mode, can allow for decreased inter- action forces and true atomic-scale resolution. According to Ohnesorge and Binnig [31], the four keys to true atomic-scale resolution are as follows: using sharp tips, imaging in solution, sys- tematically regulating forces, and, when possible,66 PA, Maurice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) 57: (c) Fig. 7, Sequence of images showing anomalous topographic features caused by changing the tip shape while imaging the (G01) surface of single-crystal hematite immersed in citric acid ‘The steps which appear to form along the edges of the large island are actually artifacts of multiple tips. Images were collected at times (a) 12:08, (b) 12:15, and (¢) 12:20, all with rightto-let scan. An image collected with a reversed scan direction exhibited steps on the left side of the island (not shown here), imaging in the attractive-force regime. Images collected in air or with relatively high forces may still be valuable in that the observed structures should be related to crystallographic parameters, even though individual atoms are not accessible. However, previous work on molecular to atomi scale imaging of mineral surfaces (see, for example, Refs. [2] and [58]) should be re-evaluated and perhaps improved upon with the help of thest guidelines. Clear, regular, repeatable atomic-scale images are needed to evaluate atomic-scale surface struc- ture, Unfortunately, atomic-scale AFM imaging is at present extremely difficult. Thus, many noisy, irregular images have been published. One common problem is that images are often pub- lished containing just one isolated regular unit cell, or a small set of unit cells, surrounded by irregular features which may be due to either noise or non- crystalline material. For example, a recent manu- script by Nishimura et al. [59] on Li sorption to muscovite shows only one, or at best a few, struc- tured <1 nm-on-a-side array(s), within a 5 nm-on- a-side image. The authors interpret the one or several ordered areas as containing single Li* ions surrounded by six oxygen atoms. The rest of the image shows noisy, irregular structures. The image was collected in solution, and with forces of only 10-100 pN. However, imaging in solution and with small forces is not always adequate to produce interpretable images. Reproducibility, both within a single image and from one image area to another, is essential. The AFM community needs to produce a set of standards to ensure that atomic-scale structures are “real”, while at the same time acknowledging that, as a local technique subject to noise and other artifacts, a statistically signifi- cant sampling may be difficult to obtain. AFM studies of atomic-scale surface structure should include imaging at numerous locations, on a number of samples or sub-samples, with a variety of tips, in different media, and with systemati variation of imaging forces. Feedback oscillations with a periodicity that could be confused with atomic-scale structure can occur if the gains are set too high. It is therefore important to confirm that apparent atomic-scale structure is “real” by systematically altering scan rate, scan size, scan angle and direction, and gains. Although atomic- scale imaging is time-consuming, such painstaking work is required. Publication of unfiltered images and transform plots is important for proper imagePA, Mawrice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) $775 6 interpretation. Two-dimensional fast Fourier- transform filtering can remove noise without intro- ducing extra spots, but unfiltered images are needed to evaluate the reliability of the data. Transform plots succinctly summarize data, and are useful for determining artifacts such as drift and double-tip effects. Combination of AFM with other techniques such as STM, LEED, XPS, XRD. and TEM is crucial, especially considering that mineral surfaces tend to be heterogeneous and that structural data may be ambiguous. There is a critical need within the AFM community to address sampling statistics, and to set standards for image quality and reproducibility. Finally, it may be necessary to slow down the race to publish until high quality, reproducible images are col- lected, with careful attention being paid to the details of the image collection and interpretation processes. 4.2. AFM as a probe of double-layer forces The use of AFM as a probe of doubl forces is one of the most exciting developments from the standpoint of mineral-water interface geochemistry. Prior to AFM development, the most widely regarded technique for measurement of surface forces was the Istaelachvili-Adams force apparatus, which works by measuring the force between two cylindrical macroscopic surfaces. Isrealachvili_ and Adams [60] measured the forces between two muscovite sheets at close approach, in solutions of different pH and ionic strength. A limitation of their experiments was the use of multiple beam interference to measure sur- face roughness. Visible light interference is prob- ably not sensitive enough to detect small-scale surface features of limited lateral extent, eg. ultrafine particles and small pits. Johnsson et al. [61] showed that muscovite may develop small etch pits and other surface features rapidly upon exposure to aqueous solution, Additionally, musco- vite is brittle and may form fine fractures upon bending (into cylindrical form). AFM circumvents these problems because surface microtopography may be measured on the nanometer scale, although sample drift makes it difficult to define the exact area of measurements. Tip-sample interaction forces can be measured by recording force curves, wherein the deflection of the cantilever is monitored as the tip approaches the sample, the tip and sample come into contact with one another, and the tip is subsequently retracted. Ducker et al. [7,8] succeeded in probing the forces between a tip consisting of a silica sphere and a planar silica surface oxidized to depth of about 30nm, immersed in solutions of different ionic strength. The authors collected force-versus- tip-sample displacement data in solutions of ionic strength 10°10"! M. To convert these measure- ‘ments to force-versus-distance data, zeros of force and distance had to be defined. The zero of deflec- tion was chosen as where the deflection was con- stant, ie. with the spherical tip and sample far apart, The zero of distance was defined based on the point at which the photodiode output became a linear function of displacement of the sample, ie. where the spherical particle was in contact with the surface. The resulting force-versus-distance curves were generally in agreement with DLVO theory. However, deviations were observed at very short distances. The authors noted that these devia~ tions could be due to hydration forces, but that the potential effects of surface roughness could complicate the interpretation. Weisenhorn et al. [9] recorded force-versus- distance curves for interactions between a SiNy tip and mica in KCI solutions at pH~6.6 and ionic strengths of 0.1-30 mM. They found that the force curves showed repulsive behavior in the non- contact regime, presumably owing to double-layer forces. The repulsion decayed exponentially with increasing distance, as expected from double-layer theory. Plots of the Debye length (1/4) versus the inverse square root of the concentration from 0.1 to 30mM_ KCI showed that 1/ decayed with increasing ionic strength, as expected from double- layer theory; the measured slope of 0.308 compared favorably with the predicted value (0.305). Radmacher et al. [62] used laterally resolved force curves to study the adsorption of organic (lysozyme) molecules adsorbed onto mica, They showed that the adhesion forces between the SisNy tip and the mica surface were different from the forces between the tip and the adsorbed lysozyme molecules. These results demonstrate the potential6 P.A, Maurice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) 57-75 for development of a form of AFM based on force- curve characteristics. Manne et al. [19] utilized non-contact-mode AFM imaging, based on double-layer repulsion between tip and sample, to image surfactant hemimicelles on highly oriented pyrolytic graphite. Hence, the double layer can be utilized to image delicate structures. Currently, the greatest challenges facing AFM studies of double-layer forces include: difficulties in defining a good zero position for measurement of tip/sample displacement, tendency for drift to alter the measurement location, and uncertainties in distance (height) measurements at very close tip/sample approach. Conducting good force- versus-distance curve measurements requires care- ful precision. Approach-withdrawal speed must be kept to a minimum so that hydrodynamic forces resulting from the displacement/replacement of water between the tip and the sample do not overshadow electrostatic forces. 4.3. In situ imaging of surface microtopography AFM is being used increasingly as a technique for imaging mineral surfaces immersed in aqueous solution, over the course of dissolution, precipita- tion, and heterogeneous nucleation reactions [10-13,15-17,63,64]. While macroscopic studies provide information on overall reaction rates and products, AFM provides complementary informa- tion on the role of surface heterogeneity. AFM permits direct measurement of surface-controlled growth and dissolution rates by providing three- dimensional data on changes in microtopography. In-situ AFM has the perhaps unique ability to detect different processes, such as dissolution and secondary phase formation, occurring simulta- neously on a mineral surface. Although forces are minimized in solution, damage of the sample surface can occur, especially during small-region scans (<1 ym) and when soft features are present; for example, over the course of dissolution or during growth of an amorphous precipitate, Fig.8 shows an example of scan- induced sample erosion during a dissolution experiment [17]. In this experiment, an etch pit was observed to nucleate and grow on the basal- plane surface of hematite immersed in citric acid, Fig. 8. Example of scan-induced erosion of a sample surface during a small-egion scan. The pit shown formed during dissolution of specular hematite in citeic acid, and was originally hexagonal in shape. However, following small-region (< 1 ym) scans, the pit shape became deformed and edges of the pit began to align with scan directions. The arrow shows a region that is being actively eroded by the tip. The original pit shape was hexagonal. However, near the end of the experiment, the scan area was decreased (from about 10 im on a side to <1 um) to the region immediately surrounding the pit. During this procedure, the pit edges began to align with the scan direction (Fig.8) and the image frame, suggesting that scan-induced erosion occurred during the small-region scans. Scan- induced erosion also was observed during small- region scan imaging of the growth of “active Cr-hydroxide precipitate on the surface of hematite [64,65]. Marchant et al. [66] showed that 1-2 jim scans disturbed the structure of von Willebrand Factor (a blood plasma glycoprotein) sorbed to mica, whereas 8pm scans did not produce a noticeable disturbance. The reason for the enhanced small-region scan erosion is currently under debate. It has been suggested that enhanced erosion during small-region scans is a result of the rapid scan rate needed to compensate for drift at high resolution [49]. However, slowing the scan rate does not always resolve the problem [67] In-situ growth experiments often show isolated holes or pits that appear on the sample surface and remain stable over long periods of time, even when several steps migrate over the holes [10,11,15]. These features have been observed during both small- (<1 jm) and large-region (>S um) scans. Whether or not they are scanP.A, Maurice!Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) 57-75 6 induced, perhaps by the frictional mechanisms proposed by Delawski and Parkinson [45], as described above, remains to be determined. Scan-induced damage often results in features that align with the scan direction and the image frame. Reaction rates may appear to vary from a small-scan (¢.g. <1 ym) to a large-scan area (e.g. 16 um). To evaluate scan-induced artifacts, in-situ reaction studies should include careful calibration and monitoring of forces, systematic alteration of sean angle, periodic disablement of the y-scan direction, periodic change in scan size, and occasional tip withdrawal and re-engagement. Additionally, scan-induced erosion should be sus- pected whenever there is evidence of the tip sticking to the sample surface, e.g. when the zero of height is reset, requiting application of a fatten filtering procedure. Finally, it should be understood that surface erosion may not be limited to small-region scans, particularly when the sample surface is soft and easily deformable or when imaging forces have not been minimized. 4.4. Imaging particulates Although single, centimetre-scale crystals can be used in the image evolution of surface microtopog- raphy over the course of reaction on surfaces immersed in solution, such in-situ experiments may suffer from two problems: (1) inability to obtain statistically meaningful sampling because such in situ experiments require painstaking set-up and lengthy experimental sessions, and (2) decoupling of the images from macroscopic sorption, growth, or dissolution data. For highly soluble or fast- reacting surfaces, such as calcite, these problems may be overcome relatively easily. An example is provided by the work of Dove and Hochella [12] on calcite precipitation mechanisms and inhibition by orthophosphate. However, for more slowly reacting surfaces characteristic of most aluminosili- cate and oxide minerals, imaging of particulate material before and after reaction may at present be a more practical technique. Imaging of particles also may be important for structural determin- ations of common clay minerals, and for measuring particle thicknesses and aspect ratios. Friedbacher et al. [21] showed that particulate material, in the form of powdered clam shells and CaCO; powder, could be imaged by pressing the powdered samples into pellets, They loaded approximately 1g of ground powder into a KBr die (Perkin Elmer) and pressurized at 5 x 108 Pa for 15 min using a Carver laboratory press. This procedure resulted in pellets 13 mm in diameter and 1.5mm in thickness. This technique has the disadvantage that particles are often overlapping so that single particles cannot be imaged in their entirety. Additionally, edges of particles may stick up, causing tip convolution artifacts A substrate is required to image isolated, single particles. Choice of substrate material is of para- mount concern, The scanning-probe microscopy ‘community was alerted to the potential dangers of substrate artifacts when highly publicized images of supposed DNA on a graphite surface were later shown to resemble strands of graphite substrate material [68,69]. Although some specialized applications may require highly specialized substrates, in general a good substrate material needs to adhere to the following criteria: (1) the substrate should be flat and regular on a scale greater than the scale of the materials of interest; (2) if the materials to be imaged are to be deposited from solution, then the substrate must be unreactive; (3) the substrate must be readily imaged, and have consistent nanometer- to micron-scale topography; (4) the atomic-scale structure and microtopography of the substrate should be distinguishable from that of the material of interest; (5) the substrate should not be easily deformed by the scanning process; (6) particles should adhere sufficiently well not to be dislodged by the imaging process, A variety of substrate materials have been uti- lized, including graphite (for STM imaging [70}), muscovite (22,23,71,72], epitaxial gold on musco- vite [73], and Mylar* film [2]. Muscovite, the mostly commonly used substrate, has the advan- tage of containing large molecularly flat areas and frictional properties similar to those of clay miner- als, However, it exhibits two major drawbacks. First, muscovite is reactive in water [74], and muscovite surfaces exposed to water for even short periods of time may contain etch pits and second- ary precipitates [61]. Second, muscovite surface70 PA. Maurice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) 57-73 structure resembles the surface structures of clay minerals. Ultrafine particles of muscovite caused by imperfect cleavage may be confused with the particles of interest, and molecular-scale imaging cannot be used to differentiate between the sub- strate and sample materials. Nevertheless, musco- vite is often the substrate of choice for imaging environmental particle. Reliable imaging of particles may be performed by syringe-filtering suspensions onto Nucleopore® polycarbonate (pc) membrane filters, and allowing the filter membranes to air-dry under a protective cover [17]. The key is to deposit so few particles that their presence can barely be detected by eye on the filter membrane surface. Too high a particle density results in a sample surface that is too rough, with overlapping particles that are difficult to image. Advantages of pe membrane substrates include: (1) the presence of pores which can be used for reference, (2) the dual use of filter mem- branes to separate solid from solution, (3) the vastly different nanometer- to atomic-scale struc- ture (organic filaments versus mineralogic), and (4) a slightly sticky surface which helps particles to adhere. A disadvantage is that the pe membrane surface is not as flat as the surface of mica, and thus is not as reliable for particle thickness mei surements. Also, the frictional properties of the filter paper may be different from the frictional properties of mineralogic specimens, leading to frictional artifacts such as inverted topography. Frictional artifacts appear to be less pronounced when using TMAFM than conventional (contact- mode) AFM. Even using low particle densities and pe mem- brane filters, imaging of particles often proves difficult. The tip convolution problems described above need to be considered, Additionally, particles can be dislodged from the surface, resulting in noisy images and potential damage to the tip. When imaging particles, it is generally best to start with a scan region less than 1m on a side, and to slowly increase the scan size. Imaging with a very slow scan speed (<1 Hz) also can prove helpful. Scanning at 90° may result in less drag between the tip and the sample. In general, TMAPM is much more reliable than contact-mode AFM for particle imaging. Finally, imaging par- ticles within a few days of sample preparation generally works best because particles tend to detach from substrate materials with time. Fig. 9 shows a TMAFM image of hematite par- ticles following two-week exposure to obligate aerobic bacteria. Experimental methods are described by Hersman et al. [75]. The bacteria specimen was a soil isolate which, by fatty acid analysis performed by Dr. Myron Sasser of Microbial ID. Inc. (Newark, DE), was determined to be most similar to Pseudomonas stutzeri. Priot to reaction, hematite particles tend to have regular hexagonal outlines. Following two-week exposure to the bacteria, many particles showed no obvious dissolution features. However, a small percentage of particles showed extreme etching [76]. Such severe etching is apparent on the particles shown in Fig. 9; only a few of the hexagonal edges remain. Small (~1 jim) elongated blebs present on the crystal surfaces may be bacterial remains. These experimental results emphasize the importance of devoting substantial time to AFM studies. AFM is a localized surface technique, but mineral-surface structure, composition, and reactivity are typically heterogeneous. Imaging natural field samples presents a chal- lenge to the microscopist, because natural samples tend to be particularly heterogeneous. AFM does not provide chemical data; hence, minerals must be identified primarily based on morphology. Fig 9. TMAFM image in air of hematite particles (on pe membrane Biter substrate) following exposure for 2 weeks to obligate aerobic Pseudomonas bacteria. Severe etching is apparent. The hematite sample was provided by L. Hersman (Los Alamos National Laboratories)PA. Maurice{Colloids Surfaces A: Physicochem. Eng, Aspects 107 (1996) 57-75 n However, morphology alone can lead to ambigu- ous results. Even atomic-scale imaging may not be conclusive because the crystal structures of many soil minerals are similar. The key to working with field samples is to take a systematic approach. First, it is essential to compile as many images as possible of well-characterized synthetic and natural samples to use as a catalog for particle identifica- tion. Second, field samples should be characterized as fully as possible by XRD, and other chemical and physical analytical techniques. Third, tech- niques for isolating different components can greatly simplify image interpretation if they are utilized cautiously. Given the high resolution capa- bility of AFM, isolation techniques that work well at lower resolution may induce artifacts or leave remanant materials detectable by AFM. Finally, AFM should be compared with other microscopies such as SEM and TEM because the different techniques provide complementary information. As part of a systematic approach to developing AFM applications to field studies, TMAFM was applied to isolated, well-characterized soil hydrous Fe(II) oxides from Typic Hapladulf soils from south-eastern Ohio [77]. These samples had been imaged previously by TEM, as reported by Bigham et al. [78]. Fig. 10 shows TMAFM images of high- gradient magnetic separates from the <0.2 am. fractions of B and C horizons. In accord with TEM images [78], hematite particles in the 2C1 horizon showed well-formed pseudohexagonal crystals (Fig. 10(a)); whereas hematite particles in the 2Bt2 horizon showed rounded crystals with large dissolution cavities indicative of dissolution and weathering (Fig. 10(b)). Some particles showed tip/sample convolution artifacts. However, in gene- ral the TMAFM images provided greater detail than the TEM images, especially on overlapping grains and aggregates (Fig. 10(b)) 4.5. Clay particle morphology and thickness measurements Blum [24] used contact-mode AFM to analyze smectite/illite particle morphology. The clay par- ticles were deposited from suspension on freshly cleaved mica, which gives a flatter background surface than pe membrane filters. After deposition, (a) (b) Fig. 10. TMAFM images of high-gradient magnetic separates, from the <0.2 4m fraction of soil samples. (a) A well-formed, pseudohexagonal hematite crystal typical of the 2C1 horizon. (b) A rounded hematite crystal, which contains a large dissolution cavity, from the 2B12 horizon. Rounding and etching ate indicative of soil weathering which has proceeded more thoroughly in the B horizons than the underlying C horizons. These images are in accord with previously published ‘TEM images [78]. The samples were provided by J. Bigham (Ohio State University). the solution was allowed to evaporate, leaving the particles attached to the surface. Blum [24] found that ultrasonic dispersion of the suspension was necessary t0 isolate individual particles. With care- ful attention to sample preparation and imaging, he concluded that AFM can be used to determine the heights of clay particles with an accuracy of +1.5A for particles of thickness in the order of several nanometers and xy-dimensions of less than 100 nm. Height accuracy for thicker and/or less laterally extensive particles was reported to be +3,0nm, Particle thickness measurements were found to agree well with measurements on the same samples by TEM and XRD. Blum [24] also concluded that for imaging vertical features larger than about 20nm, the tip shape (angle) limitsn P.A, Maurice/Colloids Surfaces A: Physicochem, Eng, Aspects 107 (1996) 57- resolution, whereas for vertical features of less than about 201m, the radius of curvature of the tip limits resolution, Fig. 11 shows TMAFM images from a study {79] comparing a poorly crystallized Clay (a) (b) (c) Fig 11. TMAFM images of Georgia Kaolinite standards deposited on pe membrane filters and imaged in aie (a) Well- cexystallized standard Kga-1, showing nicely hexagonal erystals, (b) Close-up of a hexagonal pit on the basal-plane surface of a Particle of standard Kga-l. c) Close-up along the edge of a particle of poorly-crystalline standard Kga-2, showing that the poorly-crystalline particles are generally rounded, with rough outlines. Hexagonal crystals are observed only rarely, TMAFM, images by Q, Zhou (Kent State University, Minerals Society Georgia kaolinite standard (Kga-2) with a well-crystallized CMS Georgia kaolinite standard (Kga-1b). Preliminary samples were prepared by adding a small amount of clay to deionized water, stirring for 5 min, and im- mediately syringe-filtering onto pe membrane filters. For thickness-to-edge-length measurements, some particles were also deposited on muscovite. The images shown in Fig. 11, and all TMAFM images presented herein, were collected using Nanosensors® n "silicon integrated tips Both clay standards showed some aggregate particles. Since no dispersing technique was used, this was not unexpected. Aggregates of well- crystallized samples consisted of hexagonally shaped grains of uniform size, showing “books- heets” packeting along the c-axis. Aggregates of poorly-crystallized samples contained differently sized crystals with rough surfaces and outlines. Individual grains of well-crystallized kaolinite gen- erally had regular hexagonal outlines, and hexago- nal edge lengths showed a bimodal distribution centered at 210 nm and 425 nm. The average ratios of thickness to hexagonal edge length (t/a) of small and large grains were found to be 0.33 and 0.21, respectively. Crystallographicaily controlled sur- face structures were often present on particle sur- faces. For example, Fig. 11(b) shows a hexagonal pit structure imaged on the basal-plane surface of one particle, Most individual particles of poorly- crystalline kaolinite were small, most less than 300 nm in diameter. Good hexagonal shapes were rarely observed; in general, particle edges were rounded and surfaces contained irregular massive surface features (Fig. 11(0)) 46. TMAFM imaging of humic substances Humic substances are some of the most elusive materials of interest to environmental scientists. Their structure, chemical composition, and chemi- cal reactivity vary not only from one environmental setting to another, but also with parameters such as pH, ionic strength, and metal concentrations. As polydisperse mixtures of molecules, humic substances tend to be extremely difficult to characterize. ‘A number of researchers have used SEM andP.A. Maurice/Colloids Surfaces A: Physicochem. Eng. Aspects 107 (1996) 7-75, TEM to determine the morphologies of humic substances as a function of pH, ionic streng and/or concentration [80-84]. Stevenson and Schnitzer [82] used TEM to determine the siz. shape, and degree of aggregation of soil humic substances deposited on mica by rapid freezing and drying of dilute aqueous solutions. After drying, the samples were shadowed with platinum and coated with carbon; the resulting humic acid fulvic acid (HA/FA) replicates were deposited on TEM grids for imaging. Five different types of structures were observed: 9-50nm diameter discrete spheroids, flattened aggregates of spher- oids, Jinear-chainlike assemblies of aggregates, flattened filaments, and perforated sheetlike struc- tures. Stevenson and Schnitzer [82] reported that humic substance concentration in the original solu- tion determined the predominant type of structure. Solution pH (3.5, 7.0, 100) appeared to have no effect on the types of structures observed By depositing FA on freshly cleaved muscovite, we are working to expand upon Stevenson and Schnitzer’s experiments, with AFM. AFM has a distinct advantage over TEM in that the samples do not need to be freeze-dried, coated, or subjected to vacuum, Rather, several drops of a soil FA (NaOH extract from a muck soil underlying a freshwater wetland in the New Jersey Pinelands) are simply allowed to air-dry on a muscovite surface, and the resulting samples imaged by TMAFM [85]. For the images shown in Fig. 12, the FA concentration was 50 mg |", the inherent solution pH was 4.0, and no electrolyte was added. The results were similar to those from Stevenson and Schnitzer’s [82] TEM images for samples at high concentrations, suggesting that the features seen in the TEM images were not merely artifacts of the sample preparation procedure. As observed in the TEM images, a perforated sheetlike structure was the predominant apparent structure. TMAFM, images (Fig. 12(a)) further showed a fibrous net- work present within the perforations, and several aggregates of spheres. Branched-chainlike struc- tures were also apparent at some locations on the surface (Fig. 12(b)). Hence, we observed many of the features noted by Stevenson and Schnitzer [82], all on the same sample, The presence of several different features may be due to concen tration gradients upon drying, to variable FA (b) Fig. 12. TMAFM images of soil fulvic acid deposited on ‘muscovite and imaged in air. (a) Close-up in the region of several perforations, showing fibrous network and spheroidal structures within and around the perforations. (b) Close-up of a branched-chain structure consisting of agaregated spheroids. These features are similar to those observed with TEM [82] ‘on soil humic and fulvic acids deposited on muscovite. Images bby K. Namnesnik-Dejanovie (Kent State University) composition, o to inhomogeneous structure, com- position, or microtopography of the underlying muscovite surface One potential problem with this research is the uuse of muscovite as a substrate, Muscovite has been shown to form a variety of different surface structures upon reaction with deionized water and subsequent drying [61 ]. For example, contaminant ultrafine particles may be present, and cannot be distinguished from individual spheroidal FA par ticles in Fig. 12(a). However, extensive imaging of sample blanks strongly suggests that the main features shown in Fig. 12 are FA. Ongoing research is also aimed at imaging sorbed HA/FA in-situ, as a function of pH, ionic strength, organic-acid con- centration, and sorbent mineralogy. Such in-situ"4 P.A. Maurice/Collods Surfaces A: Physicochem. Eng. Aspects 107 (1996) 57-75 experiments are important because the morphol- ogy of HA and FA materials might change upon drying. Moreover, in-situ TMAFM should assist, in distinguishing humic substances from structures on the underlying mineral substrate. 5. Conclusions The most important guideline for AFM use is to approach each study systematically, imaging in a variety of modes and under different conditions so that potential artifacts can be detected. With careful documentation, AFM is becoming an increasingly versatile and reliable technique. AFM use in mineral-water interface chemistry has only just begun; new frontiers promise to make AFM not only an imaging device, but also a dynam- ic research tool for understanding interfacial phenomena. Acknowledgments ‘The author's research has benefited greatly from discussions with many colleagues, especially M.F. Hochella, Jr., C.M. Eggleston, G. Sposito, G.A. Parks, and AE. Blum. Research on microbial interactions with iron oxides is being conducted in collaboration with G. Sposito, L. Hersman, and J. Forsythe. J. Bigham kindly provided soil samples. Graduate research assistants Q. Zhou and K. Namjesnik-Dejanovie provided images of kaolinite samples and fulvic acid samples, respec- tively. Research was conducted as part of an NSF Graduate Fellowship, and with funding from the Petroleum Research Fund of the American Chemical Society grants #28636-G2 and 25997- AC2, the National Science Foundation, grant HEAR-940-6396, and Los Alamos National Laboratories. References [1] G. Binnig, CF. Quate and Ch. Gerber, Phys. Rev., 56 (1986) 930, [2] H. Hartman, G. Sposito, A. Yang, S. Manne, SAC, Gould and P-K. Hansma, Clays Clay Miner, 38 (1990) 337, [3] MF. Hochella, Jr, CM. Eggleston, ¥.B, Elings and MS. Thompson, Am. Miner. 75 (1990) 723 [4] A. Graz, S. Manne and P.K. Hansma, Science, 51 (1991) 1348. [5] P.A. Johnsson, CM. Fggleston and M.F. Hochella, J, Am. Miner, 76 (1991) 1442. [6] F. Ohnesorge and G. Binnig, Science, 260 (1993) 1451, [7] W.A. Ducker, TJ, Senden and RM. Pashley, Nature, 353 (1991) 239. [8] WA. Ducker, TJ. Senden and RM. 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