WO2009001220A2 - Functionalization of microscopy probe tips - Google Patents

Abstract

The invention comprises a method of functionalizing scanning probe microscope (SPM) tips to image and/or measure interactions between surfaces, including the surfaces of inorganic, organic-inorganic hybrid, organic, magnetic/conductive, hard coatings and biological materials. The invention further comprises the use of atomic layer deposition (ALD) to functionalize SPM tips.

Description

FUNCTIONALIZATION OF MICROSCOPY PROBE TIPS

[001] This application claims priority to U.S. Patent Application Nos. 60/929,399, filed June 26, 2007; 60/935,008, filed July 23, 2007; and 61/039,890, filed March 27, 2008, the contents of which are incorporated herein by reference. BACKGROUND AND SUMMARY AT THE INVENTION

[002] The invention relates to the functionalization of scanning probe microscope (SPM) probe tips to study interactions between inorganic, organic- inorganic hybrid, organic, and/or biological materials. The invention also relates to the functionalization of atomic force microscope (AFM) and magnetic force microscope (MFM) probe tips. The invention further relates to the use of atomic layer deposition (ALD) as the method to functionalize these tips.

[003] The invention also relates to the use of ALD to deposit organic and/or biomolecules on probe tips, and the use of these tips to image sample surfaces and study the interactions between the material deposited on the tip and a sample surface.

[004] Molecular biologists today do not have effective tools to image realtime cellular and sub-cellular "topo-dynamics." Instead, researches often image cells that are fixed and prepared for microscopy. Advanced molecular imaging techniques that utilize fluorochromes, antibodies, and probes to analyze molecular interactions are almost always restricted to imaging of biological processes that have been arrested by fixation. Therefore, our biological knowledge is based in part on distorted still images of the dynamic situations that make up life. The study of real-time topo-dynamics of molecular interactions that take place in and on cells will dramatically increase our understanding of cellular and sub-cellular functions, clearing the road for new advances in biological, biomedical, and biomaterial sciences.

[005] The invention thus further relates to a method for investigating, in real-time, the topo-dynamics of a biological surface using an SPM equipped with a probe comprising a cantilever and a tip coated via ALD with an organic and/or inorganic and/or organic- inorganic hybrid and/or magnetic/conductive and/or hard coated and/or biomolecular film. Scanning Probe Microscopy

[006] SPM is the branch of microscopy used to image and characterize sample surfaces at the nanometer scale with a physical probe. The probe scans and interacts with the surface to measure some property of that surface. SPM is a broad term used to describe various imaging applications. The type of interaction measured between the probe tip and the sample surface determines the type of SPM technique required.

Atomic Force Microscopy

[007] AFM images sample surfaces. It can reveal structures precisely up to sub-nanometer resolution in three dimensions. For example, in one embodiment, the AFM raster-scans a surface using a probe, as shown in Figure 1. As the probe traverses the sample surface, small changes in the height of the cantilever tip may be detected by a laser. Raster-scanning the probe across a sample surfaces generates a topological map of the surface.

[008] Atomic force microscopy has revolutionized the way in which researchers explore biological structures at the single-molecule level. See B. P. Jena and J. K. Hόrber, "Atomic Force Microscopy in Cell Biology" in Methods in Cell Biology, San Diego: Academic Press, 2002. It can provide three-dimensional views of samples with minimal sample preparation. See A. Engel and D.J. Muller, Nature Structural Biology, 7(9): 715-18 (Sept. 2000). Compared to the conventional instruments for studying sample surfaces, such as the profilometer, the AFM has a much sharper tip and small loading force. This may result in improved lateral resolution of surface images.

[009] In one embodiment, atomic force microscopy may allow researchers to image samples under more natural aqueous conditions. In contrast, microscopic techniques relying on electron scanning (SEM) or electron transmission (TEM) image samples under high vacuum.

[010] The AFM not only maps surface topography, but may also be used to map surface forces. To probe surface forces, the AFM may "tap" a sample surface with the probe tip, as illustrated in Figure 2. By monitoring the cantilever deflection and by knowing the spring constant of the cantilever, one may obtain additional information about the sample, including its hardness and adhesiveness. Alternatively, the AFM may be operated with the tip in contact with the sample surface.

[011] This molecular force probing (MFP) has emerged as a powerful tool for exploring intermolecular forces and dynamics. In some embodiments, MFP's allow measurement of pico-newton (10^"12 N) forces associated with single molecules. T.E. Fisher et al., Nature Structural Biology, 7(9): 719-24 (Sept. 2000). This sensitivity allows the AFM to probe the molecular basis of biological phenomena and properties as diverse as molecular recognition [see B. Samori, Chemistry (Weinheim an der Bergstrasse, Germany) 6(23): 4249-55 (Dec. 1 , 2000); E. L. Florin et al., Science, 264(5157): 415-17 (Apr. 15, 1994)]; protein folding and unfolding [see A.F. Oberhauser et al., Nature, 393(6681 ): 181-85 (May 14, 1988)]; DNA mechanics [see M. Rief et al., Nature Structural Biology, 6(4): 346-49 (Apr. 1999)]; and focal cell adhesion [see M. Benoit et al., Nature Cell Biology, 2(6): 313- 17 (Jun. 2000)]. In one embodiment, AFM/MFP may be used to reveal the interaction between individual ligands and receptors, either on isolated molecules or on cellular surfaces.

[012] Other techniques are available for probing the interaction forces at biological surfaces and may be used in conjunction with AFM. These include the use of shear flow detachment [see J. L. Mege et al., Ce// Biophysics, 8(2): 141-60 (Apr. 1986)], surface force apparatus [see D. E. Leckband et al., Science, 255(5050): 1419-21 (Mar. 13, 1992)], biomembrane force probe [see R. Merkel et al., R, Nature, 397(6714): 50-53 (Jan. 7, 1999)], and optical tweezers [see A. Ashkin and J. M. Dziedzic, Proc. Nat'l Acad. ScL U.S.A., 86(20): 7914-18 (Oct. 1989)].

[013] AFM/MFP is a force-measuring technique that can be used to map the nanoscale lateral distribution of single molecular recognition sites on biosurfaces. Procedures to probe the forces, dynamics, and localization of molecular recognition interactions are now well established.

[014] AFM-Magnetic force microscopy (MFM) is a straightforward special mode of operation of non-contact scanning force microscope. Detection of magnetic interactions on a local scale is possible by equipping the force microscope with a magnetic probe, which then can be raster-scanned across any magnetic sample. MFM is applicable under various environmental conditions, in most cases even without requiring any special sample preparation procedure. MFM is an important analytical tool whenever the near-surface stray-field variation of a magnetic sample is of interest. See Koblischka and Hartmann, Ultramicroscopy, 97: 103-112 (2003).

[015] The recording industry became an important field of industrial application. See Rugar, Mamin et al., Journal of Applied Physics, 68: 1169-1183 (1990). MFM also exhibits some serious shortcomings and they have not been overcome so far: In the general situation, the method yields only qualitative information about the magnetic object and it is difficult to improve the resolution to values below 100 nm See Hartmann, Annual Review of Materials Science, 29: 53- 87 (1999); Wickramasinghe, Acta Materialia, 48: 347-358 (2000).

[016] Commercially available MFM tips comprise magnetic or conductive coatings with radius > 30 nm. However, reducing this radius, in order to improve the resolution of the image, usually does not provide adequate conductivity due to the changing geometry of the tip.

[017] The spatial resolution obtained by MFM may be related to both the magnetized part of the probe, which is actually exposed to the sample stray field, and to the probe-sample distance. Thus, in order to improve the lateral resolution, one may decrease the coated layer thickness. By coating tips by an atomic layer and/or several layers of conductive and/or magnetic material (Fe, Co, Ni, Nd₂Fe-_I4B, Samarium-cobalt magnets (SmCo₅), ferrites which are various mixtures of iron oxides such as Hematite (Fe₂O₃) or Magnetite (Fe₃O₄) and the oxides of other metals, Alnico which is an alloy composed primarily of aluminium, nickel and cobalt, PANiCNQ, which is a combination of emeraldine-based polyaniline (PANi) and tetracyanoquinodimethane (TCNQ)) with the described ALD technique one will increase and achieve higher resolution.

[018] MFM coated tips may be used as a tool to characterize the magnetic properties of magnetic recording media such as harddrive, memory cards, and magnetic strips, to map the surfaces of semiconductor chips for critical dimension control, or to map Dynalbeads or other magnetic spheres with antibodies attached onto cellmembranes.

SPM Tips

[019] SPM tips are probe tips comprising, for example, SiN, Si₃N₄, or SiN. The tip may be coated, for example, with gold, aluminum, or platinum. These tips may aslo be coated with other materials, for example MFM tips may be coated with a ferromagnetic film. Some probe tips are coated using imprecise techniques that may deposit multilayered coatings, which can severely affect the spring constant of the cantilevers in unpredictable ways dramatically decreasing the resolution of SPM images and maps. Moreover, these coating techniques could in many case induce a bending of the cantilever that prevent the laser from reflecting onto the detector with a correct angle. In this case, these cantilevers are useless, impossible to calibrate. Care should be taken to avoid these potential problems.

[020] Various research efforts have focused on coating the available probe tips with other materials. For example, a Langmuir-Blodgett trough may be used to deposit a monolayer of ampiphillic molecules. However, this technique exerts no atomic control of the deposited material and has limited application to inorganic materials.

[021] Some industry-specific SPM applications require a physical contact tip-hard surface, for example for quality control in the production process for silicon wafers, magnetic hard disks and tapes. When tip-hard surfaces are needed, prolonging the integrity of the probe tip is desired for long-term use, specifically for high resolution.

[022] A probe tip with a longer lifetime can reduce the cost to the company requiring tip-hard surfaces, for example in labor costs (e.g. manually changing the cantilever) and production costs (e.g. delay in the quality control process).

ALD

[023] ALD deposits films one monolayer at a time using alternating self- terminating gas-to-surface reactions. H.S. Nalwa (ed.), Handbook of Thin Film Materials, Vol. 1 , Academic Press, San Diego, CA, 2001. ALD may also be referred to as atomic layer epitaxy ("ALE") or as atomic layer chemical vapor deposition ("ALCVD").

[024] The deposition of alumina (AI₂O₃) by ALD, which is known in the art, is one non-limiting example of how the technique works. See Figure 3. First, the aluminum atoms are placed on the surface using the precursor material trimethylaluminum (AI(CH₃)₃, TMA). Most materials have a native monolayer of hydroxyl groups on the terminating surface. These hydroxyl groups serve as active sites for the reaction with TMA. When TMA is pulsed into the reaction chamber, it reacts with all sterically available hydroxyl groups and forms a new terminating layer of AI(CH₃)_X fragments. In addition, some methane (CH₄) is formed as a byproduct. This process is self-limiting in that the reaction stops when no more active surface sites are available for reaction. Excess TMA and methane may be removed from the reaction chamber under vacuum and perhaps by purging with inert gas.

[025] In one embodiment, the next step in the process is to introduce water in the gas phase. Water will react with all available methyl (CH₃) groups, forming a new hydroxyl-terminated surface and more methane. This step is also self-limiting. Excess water and methane may be removed from the system under vacuum and perhaps by purging with inert gas.

[026] The surface now once again has a monolayer of hydroxyl groups as its terminating layer. Thus, the procedure may be repeated until the desired film thickness is achieved, depositing exactly one monolayer each cycle.

[027] Different precursors may be used to produce inorganic films. Appropriate precursors include, but are not limited to, those shown in the table below.

[028] Additional precursors capable of producing inorganic films are well- known in the art. See, e.g., R. L. Puurunen, J. Appl. Phys., 97: 121301-52 (2005). Inorganic precursors include, but are not limited to BCI₃, BBr₃, B(OMe)₃, AICI₃, AIBr₃, AIMe₂CI, AIMe₂O',Pr, AIMe₂H₁ AI(OEt)₃, AI(O⁰Pr)₃, a trialkyl aluminum, GaCI₃, GaMe₃, GaCI, GaBr, GaI, GaMe₃, GaEt₃, Ga(acac)₃, Ga, GaEt₂CI, GaEt₂Me, InCI₃, InMe₃, InEt₃, ln(acac)₃, In₁, InEtMe₂, InCI, InCIMe₂, CF₃, SiCI₄, SiCI₃H₁ SiCI₂H₂, SiH₄, Si₂H₆, SiCI₃H, Si(OEt)₄, Si(O"Bu)₄, ('BuO)₃SiOH, GeCI₄, GeMe₂H₂, GeEt₂H₂, GeH₄, Ge₂H₆, SnCI₄, SnEt₄, SnMe₄, SnI₄, SnCI₄, PbBr₂, PbI₂, Pb(OAc)₂, Pb(O¹Bu)₂, Pb(thd)₂, Pb(detdc)₂, YCp₃, Y(CpMe)₃, Y(thd)₃, Cd, CdMe₂, CdCI₂, PCI₃, POCI₃, SbCI₅, Bi(Ph)₃, Bi[N(SiMe₃)₂]₃ TiCI₄, TiI₄, Ti(NMe₂)₄, Ti(NEt₂)₄, Ti(NMeEt)₄, Ti(O'Pr)₄, Ti(OEt)₄, ZrCI₄, ZrI₄, ZrCp₂CI₂, ZrCp₂Me₂, Zr(O'Pr)₂(dmae)₂, Zr(O^Bu)₄, Zr(dmae)₄, Zr(thd)₄, Zr(NMe₂)₄, Zr(NEtMe)₄, Zr(NEt₂J₄, HfCI₄, HfCI₂[N(SiMe₃)₂]₂, HfI₄, Hf(O^Bu)₄, Hf(O^fBu)₂(mmp)₂, Hf(mmp)₄, Hf(ONEt₂)₄, Hf(NEt₂J₄, Hf(NEtMe)₄, Hf[N(SiMe₃)₂]₂CI₂, Hf(NOs)₄, VOCI₃, VO(C₁Pr)₃, VO(acac)₂, Nb(OEt)₅, NbCI₅, TaF₅, TaCI₅, TaBr₅, TaI₅, Ta(OEt)₅, Ta(NMe₂)₅, Ta(NEt₂),, Ta(NEt)(NEt₂)₃, Ta(N^fBu)(NEt₂)₃, Ta(N^fBu)(NEtMe)₃, CrO₂CI₂, Cr(acac)₃, MoCI₅, WF₆, WOCI₄, WF_xO_y, W(N'Bu)₂(NMe₂)₂, Mn(thd)₃, MnCI₂, Mn, Fe(acac)₃, FeCI₃, Fe(thd)₃, Fe(^BuAMD)₂, Ru(CpEt)₂, RuCp₂, Ru(Od)₃, Ru(thd)₃, Co('PrAMD)_2, Co(acac)₃, Co(thd)₂, lr(acac)Ca(thd)₃, NiCp₂, Ni(acac)₂, Ni(apo)₂, Ni(dmg)₂, Ni('PrAMD)₂, Ni(thd)₂, Pd(thd)₂, Pd(hfac)₂, Pt(CpMe)Me₃, Pt(acac)₂₎ Cu(acac)₂, Cu(thd)₂, Cu(hfac)₂, CuCI, Cu ('PrAM D), ZnCI₂, ZnMe₂, ZnEt₂, Zn(OAc)₂, Zn, Zn[N(SiMe₃)₂], HgMe₂, Mg, Mg(Cp)₂, Mg(thd)₂, Ca(thd)₂, CaCp₂, Sr(Cp'Pr₃)₂, Sr(thd)₂, Sr(methd)₂, Sr(CpMe₅)₂, Ba(CpMe₅)₂, Ba(thd)₂, ScCp₃, Sc(thd)₃, La[N(SiMe₃)₂]₃, La(¹PrAMD)₃, La(thd)₃, Ce(thd)₄, Ce(thd)₃phen, Pr[N(SiMe₃)₂]₃, Nd(thd)₃, Sm(thd)₃, Eu(thd)₃, Gd(thd)₃, Dy(thd)₃, Ho(thd)₃, Er(thd)₃, Tm(thd)_3> and Lu[Co(SiMe₃)]₂CI.

[029] ALD's unique deposition mechanism distinguishes it from other deposition or crystal growth techniques. First, ALD exhibits different growth dynamics. This is because the precursors attach only at the available surface sites, and not on top of the same precursor molecules that have already attached. The packing density of the precursors on the surface controls the growth rate. Therefore, unlike most other deposition and crystal growth techniques, ALD monolayer growth may not depend on the distribution of the precursor or rate of formation of growth steps on the crystallites forming the film.

[030] The deposition mechanism may also make ALD unique in its ability to sequentially deposit substantially uniform monolayers on all exposed surfaces. For example, this may result in conformal coverage of cantilevers, and may provide a thin film that may not significantly influence the spring constant.

BRIEF DESCRIPTION OF THE DRAWINGS

[031] Figure 1 shows the function of an AFM probe. As the probe scans a surface, small changes in the height of the cantilever tip are detected by a laser.

[032] Figure 2 shows the AFM in the force-probing "tapping" mode. The AFM probe taps the sample surface.

[033] Figure 3 demonstrates the ALD growth of alumina.

[034] Figure 4 shows the growth rate of TiO₂ from TiCI₄ and H₂O and AI₂O₃ from TMA and H₂O as a function of temperature using a pulsing sequence of 2 s of 2 s metal precursor, 1 s purge, 2 s H₂O, and 1 s purge.

[035] Figure 5 shows a force curve generated from a human osteoblast cell probed with an AFM tip that was coated with TiO₂ by ALD.

[036] Figure 6 shows a force curve generated from a sample area with little or no adhesion to an AFM tip that was coated with TiO₂ by ALD. This force curve likely reveals the interaction of the TiO₂ tip and the SiO₂ sample substrate. [037] Figure 7 is a map of an area of human osteoblast. Darker areas have less adhesion, whereas lighter area show strong adhesion.

[038] Figure 8 shows the temperature dependence of the growth rate of CaCO₃ by ALD.

[039] Figure 9 shows the quartz crystal microbalance (QCM) (also known as quarts crystal monitor (QCM)) results from a deposition using TMA and glycine in a pulsing pattern of 1 s TMA, 1 s purge, 2 s glycine, 1 s purge.

[040] Figure 10 shows the QCM results from the deposition using TiCI₄ and glycine in a pulsing pattern of 1 s TiCI₄, 1 s purge, 2 s glycine, 1 s purge.

[041] Figure 11 shows the QCM results from the deposition using TiCI₄ and 4-aminobenzoic acid in a pulsing sequence of 4 s TMA, 3 s purge, 7 s 4- aminobenzoic acid, 3 s purge.

[042] Figure 12 shows the QCM results from the deposition using TiCI₄ and 4-aminobenzophenone in a pulsing pattern of 1 s TiCI₄, 2 s purge, 1 s 4- aminobenzophenone, 1 s purge.

[043] Figure 13 shows an AFM probe tip coated with an organic functional group (A), and molecular force probing of the surface of a bone cell. B is the adhesive force between the peptide and its receptor, C. D represents another receptor that is not specific to the peptide coating.

[044] Figure 14 shows an uncoated probe tip, a conventional magnetic coated probe tip, and a probe tip coated according to the methods described herein. The geometry of the conventional coated tip is changed in comparison with the tip coated using the disclosed method.

[045] Figure 15 shows the change in appearance of an uncoated probe tip compared with a probe tip prepared according to the disclosed coating method.

[046] Figure 16 shows results from X-ray diffraction analysis used to measure TiO₂ thickness.

[047] Figure 17 shows SEM images of cantilevers demonstrating the measurement of the width of cantilevers at the pyramidal base of the tip, before and after TiO₂ deposition.

[048] Figure 18 shows SEM images of cantilever tip morphology before and after TiO₂ deposition (A= control, B= 1 nm, C=5.3nm, D=10.9nm, E=98.4nm, F=114nm of Tiθ₂). The change in tip morphology is more pronounced with tips coated with 98.4 and 114nm Of TiO₂.

[049] Figure 19 shows spring constant measurements before and after TiO₂ coating.

[050] Figure 20 shows adhesion force results between non-coated and TiO2 coated (6nm) tips versus Nunclon surface in liquid. The results show a significant difference in adhesive force for TiO₂ coated cantilevers versus non- coated on Nunclon (p<0.001 ).

[051] Figure 21 shows adhesion force results between uncoated and TiO₂ coated (6nm) tips versus collagen I surface in liquid. The results show a significant difference in adhesive force for TiO₂ coated cantilevers versus collagen I (p<0.001 ).

[052] Figure 22A shows SEM images of uncoated cantilever tips before and after 18 scans with AFM against ZrO₂. Figure 22B shows the before and after images superposed.

[053] Figure 23 shows AFM images of uncoated cantilever tip surface morphology before and after 18 scans with AFM.

[054] Figure 24A shows SEM images of TiO₂ coated cantilever tips before and after 18 scans with AFM against ZrO₂. Figure 24B shows the before and after images superposed.

[055] Figure 25 shows AFM images of TiO₂ coated cantilever tip morphology before and after 18 scans with AFM.

[056] Figure 26A shows SEM images of ZrO₂ coated cantilever tips before and after 18 scans with AFM against ZrO₂. Figure 26B shows the before and after images superposed.

[057] Figure 27 shows AFM images of ZrO₂ coated cantilever tip morphology before and after 18 scans with AFM.

[058] Figure 28A shows SEM images of AI₂O₃ coated cantilever tips before and after 18 scans with AFM against ZrO₂. Figure 28B shows the before and after images superposed.

[059] Figure 29 shows AFM images of AI₂O₃ coated cantilever tip morphology before and after 18 scans with AFM. [060] Figure 30 shows AFM images of a floppy disk scanned with an AC240 cantilever (Si, uncoated) demonstrating that the AC240 uncoated cantilever could not detect the magnetic information contained in the floppy disk.

[061] Figure 31 shows AFM images of a floppy disk scanned with a standard MFM cantilever which did demostrate that the MFM cantilever could detect information contained in a floppy disk.

[062] Figure 32 shows AFM images of a floppy disk scanned with Fe₂CoO₄ coated cantilever. The results indicate that the Fe₂CoO₄ coated cantilever had higher sensitivity to the magnetic field.

[063] Figure 33 shows AFM images of a floppy disk scanned with a RC800 PSA (SiN) uncoated cantilever demonstrating that the RC800 cantilever could not detect the magnetic information contained in the floppy disk.

[064] Figure 34 shows AFM images of a floppy disk scanned with an RC800 PSA (SiN) cantilever coated woth Fe₂O₃ which did demonstrate that the Fe₂O₃ coated cantilever had higher sensitivity to the magnetic field than the uncoated cantilever.

[065] Figure 35 shows AFM images of a floppy disk scanned with an RC800 PSA (SiN) cantilever coated woth CoFe₃O₄ which did demonstrate that the Fe₂O₃ coated cantilever had higher sensitivity to the magnetic field than the uncoated cantilever.

DETAILED DESCRIPTION

[066] Surprisingly, ALD may be used to deposit a myriad of coatings on probe tips, expanding the range of surface imaging and force-probing capabilities. The invention comprises functionalizing probe tips to image sample surfaces and to probe interactions between the tips and sample surfaces, for example for AFM and MFM applications. The invention further comprises the use of ALD as the method of functionalization. The invention also comprises the actual use of the functionalized tips to image sample surfaces and/or to probe interactions between the tips and sample surfaces. The material coating the tip and the sample surface may be the same or different, and may be inorganic, organic-inorganic hybrid, organic, magnetic/conductive, biological materials, and/or hard coatings. [067] The invention further comprises the use of ALD to produce organic and/or biomolecular films. These films may be deposited on any appropriate surface, for example on an AFM tip. A tip coated with an organic and/or biomolecular film may be used to image sample surfaces and probe intermolecular forces between the surface and the organic and/or biomolecular tip coating. Appropriate sample surfaces include any surface that may be imaged by SPM, however, biological surfaces are of particular interest.

[068] In one embodiment, ALD deposits substantially uniform films of pinhole-free materials on substrates having complex geometrical shapes. This technique of modifying probe tips may provide several advantages. For example, in one embodiment, ALD conserves the low spring constant of the probe. Therefore, it may modify tips while retaining the high resolution of the SPM.

[069] As used herein, "substantially uniform" means that there is a detectable amount of material on all exposed surfaces. "Substantially uniform" films may include, but are not limited to, films resulting from the sequential deposition of one or more monolayers of material, wherein each monolayer may be the same or different as the previous layer.

[070] The term "monolayer" a film or layer of material approximately one molecule of precursor thick. The term therefore may vary depending on the film being deposited.

[071] Examples of other embodiments of using ALD may include at least one of the following advantages. The technique is well-suited to the deposition of inorganic, organic, organic-inorganic hybrid, and biological materials. At relatively low temperatures, it can produce surfaces terminated by a desired functionality. Tailoring the surfaces of probe tips may allow one to construct and study a myriad of tip-specimen surface interactions on a molecular level.

[072] In one embodiment, a precursor molecule should not undergo reactions with itself.

[073] In order to improve and/or ensure the saturation density of the terminating hydroxyl groups on the surface of the SPM tip, the surface may be pulsed with water before depositing the initial precursor molecule. Any excess water may be purged or evacuated from the reaction chamber before pulsing the surface with the first precursor. [074] In one embodiment, the functionalization of the probe tips can be performed by deposition of one or more monolayers of different materials that either mimic bone, simple extracellular matrices, implant surfaces, or biological signal molecules (ligands).

Bone-Like Materials

[075] In one embodiment, ALD may be used to produce a film that mimics the termination of bone material by terminating in, for example, a calcium phosphate layer. Bone- or implant-like materials are those that either induce or conduct formation of new bone. Such material often have chemical structures involving sodium oxides, silicate, calcium oxides, calcium sulphates, calcium phosphates, calcium carbonates, hydroxyapatite, hydrides, vanadium, platinum, hafnium, gold hydroxide, fluorides and oxides of titanium, ferro-titanium alloys, and tantalum metals, or combination of above mention materials.

[076] For example, Ca(Cp)₂ may serve as one precursor reacting with available surface sites. Triphenyl phosphine, P(Ph)₃, may then be pulsed into the reaction chamber, forming calcium-phosphorous bonds at the surface. An oxygen source such as ozone or water may then be pulsed in to the reaction chamber to create the terminal phosphate groups.

[077] Alternatively, calcium phosphate materials may be deposited using Ca(Cp)₂ as a first precursor, followed by H₃PO₄, POCI₃, or other suitable phosphate precursors.

[078] In anther embodiment, Ca₂(PO₄)F may be deposited using the following sequence:

Step 1 : Ca(Cp)₂ + 2H₂O = Ca(OH)₂ + 2H-Cp Step 2a: 3Ca(OH)₂ + 2P(Ph)₃ = Ca₃(PO₃J₂ + 6H-Ph Step 3a: Ca₃(PO₃)₂ + O₃ = Ca₃(PO₄J₂ Step 2b: 3Ca(OH)₂ + 2POCI₃ = Ca₃(PO₄)₂ + 6HCI Step 2c: 2Ca(OH)₂ + POCI₃ = Ca₂(PO₄)(OH) + 3HCI Step 3c: Ca₂(PO₄)(OH) + NH₄F = Ca₂(PO₄)F + NH₃ + H₂O [079] Said bone material may be deposited on a probe tip. These tips may optionally be used to image sample surfaces and to study the interactions between the bone-like material and sample surfaces.

Implant-Like Materials

[080] In another embodiment, implant-like surfaces can be deposited by using materials including, but not limited to, titanium oxide (TΪO₂). TΪO2 may be deposited by using TiCI₄ and water as alternating precursors. Implant-like materials often have chemical structures involving sodium oxides, silicate, calcium oxides, calcium sulphates, calcium phosphates, calcium carbonates, hydroxyapatite, hydrides, vanadium, platinum, hafnium, gold hydroxide, fluorides and oxides of titanium, ferro-titanium alloys and tantalum metals, or combination of these materials.

[081] In another embodiment, said implant-like material may be deposited on a probe tip. These tips may then be used to image sample surfaces and to study the interactions between the implant-like material and sample surfaces.

Inorganic-Organic Hybrid Materials

[082] In another embodiment of the invention, ALD may also be used to deposit an organic-inorganic hybrid films by alternating an inorganic precursor with an organic precursor having at least one functional group. For sequential depositions of inorganic/organic precursors, the organic precursor must have at least two functional groups. An organic precursor with only one functional group may be used as the terminating layer. See WO 2006/071126A1.

[083] In one embodiment, the initial precursor used to deposit an inorganic-hybrid film may be an inorganic precursor selected from a group consisting of metal alkyls, metal cycloalkyls, metal aryls, metal amine, metal silylamine, metal halogenides, metal carbonyls and metal chelates, where the metal is selected from the group comprising Al, Si, Ge, Sn, In, Pb, alkali metals, alkaline earth metals, 3d-insertion metals, 4d-insertion metals, 5d-insertion metals, lanthanides, and actinides. [084] This precursor may be pulsed into the reaction chamber under conditions such that it reacts with the available surface sites. Any excess inorganic precursor molecules may optionally be removed from the reaction chamber by purging with inert gas or evacuating the chamber.

[085] In one embodiment, the inorganic precursor bound to the surface may then be reacted with an organic precursor by pulsing said organic precursor into the reaction chamber. The organic precursor may be an organic compound with two or more reactive substituents selected from the group comprising -OH, - OR, =O, -COOH, -SH, -SO₄H, -SO₃H, -PH₂, -PO₄H, -PO₃H, -PRH₁ -NH₂, -NH₃I, - SeH, -SeO₃H, -SeO₄H, -TeH, -AsH₂, -AsRH, -SiH₃, -SiRH₂, -SiRR¹H, -GeH₃, - GeRH₂, -GeRR'H, amine, alkyl amine, silated amine, halogenated amine, acetic anhydride, imide, azide and nitroxyl; where R and R' may be a C-M_O aryl, alkyl, cycloalkyl, alkenyl or alkynyl group. Pulsing conditions should be used such that the organic precursor reacts with the inorganic-functionalized surface forming an inorganic-organic hybrid layer. Any excess organic precursor may optionally be removed by either purging the reaction chamber with inert gas and/or evacuating the chamber.

[086] In one embodiment, the sequential deposition of inorganic and organic precursors may optionally be repeated using the same or different inorganic and organic precursors until the desired film thickness and surface termination is achieved. For sequential deposition, the organic precursor must have more than one reactive substituent.

[087] In one embodiment, an organic precursor having only one or more reactive substituent may be used to deposit the terminating layer of the hybrid film. The reactive substituent of said organic molecule may be chosen from the group comprising -OH, -OR, =O, -COOH, -SH, -SO₄H, -SO₃H, -PH₂, -PO₄H, -PO₃H, - PRH, -NH₂, -NH₃I, -SeH, -SeO₃H, -SeO₄H, -TeH, -AsH₂, -AsRH, -SiH₃, -SiRH₂, - SiRR¹H, -GeH₃, -GeRH₂, -GeRR'H, amine, alkyl amine, silated amine, acetic anhydride, halogenated amine, imide, azide, and nitroxyl; where R and R' may be a C-ι-₁₀ aryl, alkyl, cycloalkyl, alkenyl, or alkynyl group.

[088] In another embodiment, said inorganic-organic hybrid material may be deposited on a probe tip. These tips may optionally be used to image sample surfaces and to study the interactions between the bone-like material and sample surfaces.

[089] ALD has been used to produce functionalized inorganic-organic hybrid films using numerous combinations of precursors. The following examples of suitable precursors exemplify the types of precursors suitable for this technique, but are not meant to limit the scope of this invention: TMA and hydroquinone (Hq), TMA and malonic acid; TMA and terephtalic acid; ZrCI₄ and Hq; TiCI₄ and Hq; TiCI₄ and ethylenediamine. Additional inorganic precursors include, but are not limited to, BCI₃, BBr₃, B(OMe)₃, AICI₃, AIBr₃, AIMe₂CI, AIMe₂CPr, AIMe₂H, AI(OEt)₃, AI(O"Pr)₃, a trialkyl aluminum, GaCI₃, GaMe₃, GaCI, GaBr, GaI, GaMe₃, GaEt₃, Ga(acac)₃, Ga, GaEt₂CI, GaEt₂Me, InCI₃, InMe₃, InEt₃, ln(acac)₃, In₁, InEtMe₂, InCI, InCIMe₂, CF₃, SiCI₄, SiCI₃H, SiCI₂H₂, SiH₄, Si₂H₆, SiCI₃H, Si(OEt)₄, Si(O⁰Bu)₄, ('BuO)₃SiOH, GeCI₄, GeMe₂H₂, GeEt₂H₂, GeH₄, Ge₂H₆, SnCI₄, SnEt₄, SnMe₄, SnI₄, SnCI₄, PbBr₂, PbI₂, Pb(OAc)₂, Pb(O¹Bu)₂, Pb(thd)₂, Pb(detdc)₂, YCp₃, Y(CpMe)₃, Y(thd)₃, Cd, CdMe₂, CdCI₂, PCI₃, POCI₃, SbCI₅, Bi(Ph)₃, Bi[N(SiMe₃)₂]₃ TiCI₄, TiI₄, Ti(NMe₂)₄, Ti(NEt₂)₄, Ti(NMeEt)₄, Ti(O⁷Pr)₄, Ti(OEt)₄, ZrCI₄, ZrI₄, ZrCp₂CI₂, ZrCp₂Me₂, Zr(O'Pr)₂(dmae)₂, Zr(O^fBu)₄, Zr(dmae)₄, Zr(thd)₄, Zr(NMe₂)₄, Zr(NEtMe)₄, Zr(NEt₂)₄, HfCI₄, HfCI₂[N(SiMe₃)₂]₂, HfI₄, Hf(O^Bu)₄, Hf(O'Bu)₂(mmp)₂, Hf(mmp)₄, Hf(ONEt₂)₄, Hf(NEt₂)₄, Hf(NEtMe)₄, Hf[N(SiMe₃)₂]₂CI₂, Hf(NO₃)₄, VOCI₃, VO(C₁Pr)₃, V0(acac)₂, Nb(OEt)₅, NbCI₅, TaF₅, TaCl₅, TaBr₅, TaI₅, Ta(OEt)₅, Ta(NMe₂J₅, Ta(NEt₂)₅, Ta(NEt)(NEt₂)₃, Ta(N'Bu)(NEt₂)₃, Ta(N^Bu)(NEtMe)₃, CrO₂CI₂, Cr(acac)₃, MoCI₅, WF₆, WOCI₄, WF_xOy, W(N'Bu)₂(NMe₂)₂, Mn(thd)₃, MnCI₂, Mn, Fe(acac)₃, FeCI₃, Fe(thd)₃, Fe(^BuAMD)₂, Ru(CpEt)₂, RuCp₂, Ru(Od)₃, Ru(thd)₃, Co(PrAMD)_2, Co(acac)₃, Co(thd)₂, lr(acac)Ca(thd)₃, NiCp₂, Ni(acac)₂, Ni(apo)₂, Ni(dmg)₂, Ni('PrAMD)_2, Ni(thd)₂, Pd(thd)₂, Pd(hfac)₂, Pt(CpMe)Me₃, Pt(acac)₂, Cu(acac)₂, Cu(thd)₂, Cu(hfac)_2l CuCI, Cu('PrAMD), ZnCI₂, ZnMe₂, ZnEt₂, Zn(OAc)₂, Zn, Zn[N(SiMe₃J₂], HgMe₂, Mg, Mg(Cp)₂, Mg(thd)₂, Ca(thd)₂, CaCp₂, Sr(Cp'Pr₃)₂, Sr(thd)₂, Sr(methd)₂, Sr(CpMe₅)₂, Ba(CpMe₅J₂, Ba(thd)₂, ScCp₃, Sc(thd)₃, La[N(SiMe₃)₂]₃, La(¹PrAMD)₃, La(thd)₃, Ce(thd)₄, Ce(thd)₃phen, Pr[N(SiMe₃)₂]₃, Nd(thd)₃, Sm(thd)₃, Eu(thd)₃, Gd(thd)₃, Dy(thd)₃, Ho(thd)₃, Er(thd)₃, Tm(thd)₃, and Lu[Co(SiMe₃)]₂CI.

[090] Some of the functional groups imagined to undergo reactions by the ALD principle and thus suitable as functional groups to attach active molecules to the surface are described in the following. For all the proposed reaction mechanisms, it is likely that the reaction scheme in reality is somewhat shifted or different. Thus, the reaction schemes should not be interpreted limiting the scope of the invention. The main principal is that the reaction results in film formation. Therefore, the following reaction schemes serve only to show the potential of the following functional groups to form a component of a film.

[091] Concerning possible reactions between metal containing precursors and organic molecules with functional groups, organic molecules with functional groups that have some degree of acidity, i.e., that can donate a proton may be preferred. This proton will be used to complete the alkane molecule or halogen acid molecule from the inorganic precursor and let the reaction proceed. Hvdroxyl groups

[092] Hydroxyl groups (R-OH) provide an oxygen and hydrogen for a possible reaction. These may react readily with electropositive metals, including but not limited to metal alkyls and metal halides, whereby metal alkoxides and alkyl or hydrohalogen acids may be produced, respectively.

[093] Two partial reactions that may take place between a diol (HO-R-OH) and TMA ((CH₃)3AI) are given below: HO-R-OH(g) + CH₃-AI-I → CH₄(g) + HO-R-O-AI-| 2HO-R-I + (CH₃J₃AKg) → 2CH₄(g) + CH₃-AI-(O-R-I)₂

[094] Electropositive metals known to readily undergo such reactions are: Al, Mg, Si, Ti, V, and several other metals including, but not limited to, Zn, Mn, Fe, Co, and/or Cr. Ether groups

[095] Ether groups (-OR) may react and form adducts to metals in the film. These bonds may be rather weak, but may still form the basis of structure formation for films produced at low temperatures. An example of such a reaction scheme is: R'O-R(g) + CH₃-Mn-I → R¹O-R-Mn-I 2RO-R-I + (CHs)₃AKg) → 2R'-CH₃(g) + CH₃-AI-(O-R-I)₂ [096] Only one half reaction is presented, because the film formed via this reaction path may use another type of functional group in its structure to form a film in the next step. Ketone groups

[097] Ketones (R=O) may interact with metal atoms, and molecules with more than one ketone moiety may chelate metal atoms. One such example is the formation of compounds with β-ketones. An example of such visualized reaction scheme is: O=R'-R=O (g) + Mn-| → R,R'(=O)₂ -Mn-|

[098] Only one half reaction is presented, because the film formed via this reaction path may use another type of functional group in its structure to form the film in the next step. Carboxyl groups

[099] Carboxyl groups (-COOH) have the same building units as for hydroxyls (-OH) and ketones (=O).

[0100] Two partial reactions that take place between a dicarboxylic acid (HOOC-R-COOH) and TMA ((CH₃)₃AI) are given below. HOOC-R-COOH(g) + CH₃-AI-| → CH₄(g) + HOOC-R-COO-AI-I 2HOOC-R-I + (CH₃)₃AI(g) → 2CH₄(g) + CH₃-AI-(COO-R-I)₂

[0101] TMA may also react with both the =O and the -OH of a carboxyl acid.

Thiol groups

[0102] Thiol groups (-SH) may form participate in similar types of reactions as their isoelectronic hydroxyl relatives (-OH). However, metal affinity towards sulphur differs from that towards oxygen. Elements including, but not limited to, Pb, Au, Pt, Ag, Hg, and others react and form stable bonds towards sulphur.

[0103] Two partial reactions that may take place between a di-thiol (HS-R- SH) and Pt(thd)₂ are given below: HS-R-SH(g) + thd-Pt-| → Hthd(g) + HS-R-S-Pt-| 2HS-R-I + Pt(thd)₂(g) → Hthd(g) + thd-Pt-S-R-|

Sulphate groups

[0104] Sulphate groups (-SO₄H) may react with electropositive metals in ways similar to hydroxyls or ketones.

[0105] Two partial reactions that may take place between a disulphate (HSO₄-R-SO₄H) and TMA are given below: HSO₄-R-SO₄H (g) + CH₃-AI-I → CH₄(g) + HSO₄-R-SO₄-AI-I 2HSO₄-R-I + (CHa)₃AKg) → 2CH₄(g) + CH₃-AI-(SO₄-R-I)₂

Sulphite groups

[0106] Sulphite groups (-SO₃H) may react in ways similar to sulphate groups.

[0107] Two partial reactions that take place between a disulphite (HSO₃-R- SO₃H) and TMA ((CH₃)₃AI) are given below: HSO₃-R-SO₃H (g) + CH₃-AI-I → CH₄(g) + HSO₃-R-SO₃-AI-I 2HSO₃-R-I + (CH₃)₃AI(g) → 2CH₄(g) + CH₃-AI-(SO₃-R-I)₂

Phosphide groups

[0108] Two partial reactions that may take place between a di-phospide (H₂P-R-PH₂) and Ni(thd)₂ are given below: H₂P-R-PH₂(g) + thd-Ni-| → Hthd(g) + H₂P-R-PH-Ni-I 2H₂P-R-I + Ni(thd)₂(g) → Hthd(g) + thd-Ni-PH-R-|

Phosphate groups

[0109] Two partial reactions that may take place between a diphosphate (HPO₄-R-PO₄H) and TMA ((CH₃J₃AI) are shown below: HPO₄-R-PO₄H (g) + CH₃-AI-I → CH₄(g) + HPO₄-R-PO₄-AI-I 2HPO₄-R-I + (CHs)₃AKg) → 2CH₄(g) + CH₃-AI-(PO₄-R-I)₂ Amine groups

[0110] Amine groups, alkyl amines, or silated amines, or halogenated amines, may react with compounds including but not limited to SnI₂, SnI₄, PbI₂, PbI₄, CuI₂, CuI₄ or similar compounds to form perovskite-related hybrid materials as described by D. B. Mitzi (D. B. Mitzi, Progress in Inorganic Chemistry, 48: 1-121 (1999); D. B. Mitzi, Chem. of Materials, 13: 3282-98 (2001 )).

[0111] One proposed reaction mechanism is: Snl₄(g) + NH₄I-R-I → SnI₄-NH₄I-R-I SnI₄-I + NH₄l-R-H₄NI(g) → NH₄I-R-H₄NI-SnI₄-I

[0112] A redox-reaction with Sn(IV)-Sn(II) and formation of I₂(g) might be involved here. Alternatively, by using divalent halogenides, the reaction might be visualized as:

Snl₂(g) + NH₄I-R-I → SnI₃-NH₄-R-I SnI₃-I + NH₄l-R-H₄NI(g) → NH₄I-R-H₄N-SnI₄-I

[0113] In addition or in the alternative, amines may react similar to hydroxyl groups. Two potential partial reactions that may take place between a diamine (H₂N-R-NH₂) and TMA ((CH₃)₃AI) are given below: H₂N-R-NH₂(g) + CH₃-AI-I → CH₄(g) + H₂N-R-NH-AI-I 2H₂N-R-I + (CHa)₃AKg) → 2CH₄(g) + CH₃-AI-(NH-R-I)₂

[0114] Both of the H-atoms on one of the amines may react with TMA.

[0115] The following functional groups will react in a similar way: -OH, -SH, -SeH₁ -TeH, -NH₂, -PH₂, -AsH₂, -SiH₃, -GeH₃. -SO₄H, -SO₃H, -PO₄H, -PO₃H, SeO₃H, SeO₄H. In all cases where more than one H is present, the other H may be substituted by another organic group R, where R is straight and branched chain alkane, cycloalkane, an aryl group, a heteroaryl group, or a functional groups.

[0116] For precursors with more than one functional group, the functional groups need not be of the same type. Different functional groups with different reactivity may form a monolayer of organic molecules with a degree of ordering. In addition, different inorganic precursors may have different affinities for the different groups. Different organic and inorganic precursors may be used to produce various terminating surfaces.

[0117] The organic compound carrying the functional groups is not particularly limited but can be any organic molecule that can be brought into the gas phase. It is preferred that the organic precursor molecule with more than one functional group will have some form of structural or steric hindrance to prevent all of its functional groups from reacting with the same surface. For organic precursors with more than one reactive site, it is preferred that at least one reactive site does not react with the active surface sites and remains for use in a subsequent reaction. Otherwise, there are no limitations on the structure of the organic precursors. The organic molecule may influence the acidity of the protons on the functional groups. The organic compound may be a non-branched alkane, branched alkane, cyclo alkane, alkene, a monocyclic or polycyclic aromatic group, a heterocyclic aromatic group, where these compounds, in addition to the functional groups, may be substituted or not substituted with other organic groups like alkyl.

[0118] Some of the inorganic precursors that may be used to make inorganic-organic hybrid films are described below. All the suggested reactions are only illustrations of possible reactions, and are not to be interpreted as limitations. Metal alkyls

[0119] Metal alkyls and metal cycloalkyls may be rather reactive and hence may react with most organic functional groups. Examples of possible metal alkyls are: AI(CH₃)₃, Zn(Et)₂, Zn(Me)₂, and MgCp₂I. Metal halogenides

[0120] Some electropositive metal halogenides may be rather reactive and therefore may undergo reaction with many organic functional groups. Some examples are AICI₃, TiCI₄, SiCI₄, SnCI₄, Si(CH₃)₂CI₂. Metal carbonyls

[0121] Metal carbonyls may also be reactive, including but not limited to: Fe₂(CO)₉, Mn(CO)_x. Metal chelates

[0122] Reactive metal chelates may include, but are not limited to: VO(thd)₂, Mn(HMDS)₂, Fe(HMDS)₂, TiO(thd)₂, Pt(thd)₂, where HMDS stands for hexamethyl-disilazane.

[0123] Other possible chelates are beta-ketones such as acetylacetonates, fluorinated thd-compounds and ethylenediaminetetra acetic acid (EDTA).

[0124] The metal for the inorganic precursor is selected from the group consisting of Al, Si, Ge, Sn, In, Pb, alkali metals, alkaline earth metals, 3d-insertion metals, 4d-insertion metals, 5d-insertion metals, lanthanides, and actinides. Metals of particular interest may include, but are not limited to, Cu, Ni, Co, Fe, Mn, and V.

[0125] In one embodiment of the invention, hybrid films may be fabricated via ALD using TMA as the inorganic precursor and Hq and/or PhI as organic precursor(s). This may result in films of aluminium benzene oxides. Films are produced by usage of TMA-PhI, TMA-Hq, and a controlled mixture of the type TMA- PhI-TMA-Hq. The growth kinetic may be investigated using quartz crystal monitor (QCM) measurements. The films may optionally be analyzed by Fourier transformed infrared spectroscopy (FT-IR). Active surfaces terminating in hydroxy! groups on aromatics or metal alkyls are obtained by using Hq/Phl or TMA respectively as the last type of precursor.

[0126] In another embodiment of the invention, hybrid films may be deposited from TMA and one or more organic precursors that will react with the methyl-aluminum surface, including but not limited to 1 ,4-benzenedicarboxylic acid, 1 ,3-benzenedicarboxylic acid, 1 ,3,5-benzenetricarboxylic acid, and/or 1 ,2,4,5- benzenetetracarboxylic acid.

Organic Films

[0127] In one embodiment of the invention, ALD may be used to deposit organic films. As described in M. Putkonen et al., J. Mater. Chem., 17: 664 (2007) polyimide films may be grown using anhydrides and diamines. Suitable anhydrides include those with two anhydride moieties, for example:

1 ,2,4,5-Benzenetetracarboxylic anhydride (Pyromellitic dianhydride, PMDA)

[0128] Similarly, suitable diamines include those with two amino groups. For example:

Ethylenediamine, EDA 1,6-Diaminohexane, DAH

1,4-Phenylenediamine, PDA 4,4'-oxydianiline, ODA

[0129] In one embodiment of the invention, 1 ,2,4,5-Benzenetetracarboxylic anhydride may react with a diamine to form a film using ALD.

[0130] In addition, polyamide films may be deposited using an acyl chloride and a diamine. See A. Kubono et al., Thin Solid Films, 289: 107 (1996). Suitable acyl chlorides include those with two or more acyl chloride groups. For example, nonanedioyl chloride (azelaoyl dichloride (ADC)) may be used with diaminoheptane to deposit organic films.

[0131] Alternatively, polyamide films may be deposited by alternating dicarboxylic acids and diamines.

[0132] In another embodiment, ALD may be used to deposit organic films on an AFM tip. These tips may optionally be used to image sample surfaces and to study the interactions between the bone-like material and sample surfaces.

[0133] Where a precursor contains two or more functional groups, the functional groups may be the same or different. The precursor should leave a reactive site suitable for succeeding growth with another type of precursor, unless no further deposition steps are desired. For production of the final terminating surface, the requirement of two or more types of functional groups does not apply. Rather the precursor should have at least one functional group that will undergo reactions with the previous surface, and also contain the groups that should finally terminate the surface.

Biomolecular Films

[0134] In another embodiment, ALD may be used to produce biomolecular films. Biological surfaces may be constructed by using highly reactive metal precursors to link the native hydroxyl group terminating layer to the desired biological molecules.

[0135] The term "biomolecules" is intended to cover and comprise within its meaning a very wide variety of biologically active molecules in the widest sense of the word, be they natural biomolecules (i.e. naturally occurring molecules derived from natural sources), synthetic biomolecules (i.e. naturally occurring molecules prepared synthetically as well as non-naturally occurring molecules or forms of molecules prepared synthetically) or recombinant biomolecules (i.e. prepared through the use of recombinant techniques).

[0136] A non-limiting list of main groups of and species biomolecules that are contemplated as being suitable for coating probe tips in accordance with the invention is given below. Extracted biomolecules Bioadhesives:

[0137] Bioadhesives include biomolecules that mediate attachment of cells, tissue, organs or organisms onto non-biological surfaces like glass, rock etc. This group of bio-molecules includes the marine mussel adhesive proteins, fibrin-like proteins, spider-web proteins, plant-derived adhesives (resins), adhesives extracted from marine animals, and insect-derived adhesives (like resilins).

[0138] Some specific non-limiting examples of adhesives include, but are not limited to: fibrin; fibroin; Mytilus edulis foot protein (mefpi , "mussel adhesive protein"); other mussel's adhesive proteins; proteins and peptides with glycine-rich blocks; proteins and peptides with poly-alanine blocks; and silks. Cell attachment factors: [0139] Cell attachment factors include biomolecules that mediate attachment and spreading of cells onto biological surfaces or other cells and tissues. This group of molecules typically contains molecules participating in cell- matrix and cell-cell interaction during vertebrate development, neogenesis, regeneration, and repair. Typical biomolecules in this class are molecules on the outer surface of cells like the CD class of receptors on white blood cells, immuneglobulins, and haemagglutinating proteins, and extracellular matrix molecules/ligands that adhere to such cellular molecules.

[0140] Typical examples of cell attachment factors with potential for use as bioactive coating on metal hybrid -coated probe tips include, but are not limited to: ankyrins; cadherins (Calcium dependent adhesion molecules); connexins; dermatan sulphate; entactin; fibrin; fibronectin; glycolipids; glycophorin; glycoproteins; heparan sulphate; heparin sulphate; hyaluronic acid; immunglobulins; keratan sulphate; integrins; laminins; N-CAMs (Calcium independent Adhesive Molecules); proteoglycans; spektrin; vinculin; vitronectin.

Biopolvmers:

[0141] Biopolymers are any biologically prepared molecules that, under the right conditions, may be assembled into polymeric, macromolecular structures. Such molecules constitute important parts of the extracellular matrices where they participate in providing tissue resilience, strength, rigidity, integrity etc. Some important biopolymers with potential for use as bioactive coating on metal hybrid- coated cantilever include, but are not limited to: alginates; Amelogenins; cellulose; chitosan; collagen; gelatins; oligosaccharides; pectin.

Blood proteins:

[0142] Blood proteins typically contain any dissolved or aggregated protein that normally is present whole blood. Such proteins can participate in a wide range of biological processes like inflammation, homing of cells, clotting, cell signaling, defense, immune reactions, metabolism etc. Typical examples with potential for use as bioactive coating on metal hybrid -coated cantilever include, but are not limited to: albumin; albumen; cytokines; factor IX; factor V; factor VII; factor VIII; factor X; factor Xl; factor XII; factor XIII; hemoglobins (with or without iron); immunglobulins (antibodies); fibrin; platelet derived growth factors (PDGFs); plasminogen; thrombospondin; transferrin.

Enzymes:

[0143] Enzymes are any protein or peptides that have a specific catalytic effect on one ore more biological substrates which can be virtually anything from simple sugars to complex macromolecules like DNA. Enzymes are potentially useful for triggering biological responses in the tissue by degradation of matrix molecules, or they could be used to activate or release other bioactive compounds in the implant coating. Some important examples with potential for use as bioactive coating on metal hybrid -coated cantilever include, but are not limited to: abzymes (antibodies with enzymatic capacity); adenylate cyclase; alkaline phosphatase; carboxylases; collagenases; cyclooxygenase; hydrolases; isomerases; ligases; lyases; metallo-matrix proteases (MMPs); nucleases; oxidoreductases; peptidases; peptide hydrolase; peptidyl transferase; phospholipase; proteases; sucrase- isomaltase; TIMPs; transferases.

Extracellular Matrix proteins and biomolecules:

[0144] Specialized cells, e.g. fibroblasts and osteoblasts, produce the extracellular matrix. This matrix participates in several important processes. The matrix is crucial for i.a. wound healing, tissue homeostasis, development and repair, tissue strength, and tissue integrity. The matrix also decides the extracellular milieu like pH, ionic strength, osmolarity etc. Furthermore extracellular matrix molecules are crucial for induction and control of biomineral formation (bone, cartilage, teeth). Important extracellular proteins and biomolecules with potential for use as bioactive coating on metal hybrid -coated cantilever include: ameloblastic amelogenins; collagens (I to XII); dentin-sialo-protein (DSP); dentin-sialo-phospho- protein (DSPP); elastins; enamelin; fibrins; fibronectins; keratins (1 to 20); laminins; tuftelin; carbohydrates; chondroitin sulphate; heparan sulphate; heparin sulphate; hyaluronic acid; lipids and fatty acids; lipopolysaccarides.

Growth factors and hormones:

[0145] Growth factors and hormones are molecules that bind to cellular surface structures (receptors) and generate a signal in the target cell to start a specific biological process. Examples of such processes are growth, programmed cell death, release of other molecules (e.g. extracellular matrix molecules or sugar), cell differentiation and maturation, regulation of metabolic rate etc. Typical examples of such biomolecules with potential for use as bioactive coating on metal hybrid -coated probe tips include, but are not limited to: activins (Act); Amphiregulin (AR); Angiopoietins (Ang 1 to 4); Apo3 (a weak apoptosis inducer also known as TWEAK, DR3, WSL-1 , TRAMP or LARD); Betacellulin (BTC); Basic Fibroblast Growth Factor (bFGF, FGF-b); Acidic Fibroblast Growth Factor (aFGF, FGF-a); 4- 1BB Ligand; Brain-derived Neurotrophic Factor (BDNF); Breast and Kidney derived Bolokine (BRAK); Bone Morphogenic Proteins (BMPs); B-Lymphocyte Chemoattractant/B cell Attracting Chemokine 1 (BLC/BCA-1 ); CD27L (CD27 ligand); CD30L (CD30 ligand); CD40L (CD40 ligand); A Proliferation-inducing Ligand (APRIL); Cardiotrophin-1 (CT-1 ); Ciliary Neurotrophic Factor (CNTF); Connective Tissue Growth Factor (CTGF); Cytokines; 6-cysteine Chemokine (ΘCkine); Epidermal Growth Factors (EGFs); Eotaxin (Eot); Epithelial Cell-derived Neutrophil Activating Protein 78 (ENA-78); Erythropoietin (Epo); Fibroblast Growth Factors (FGF 3 to 19); Fractalkine; Glial-derived Neurotrophic Factors (GDNFs); Glucocorticoid-induced TNF Receptor Ligand (GITRL); Granulocyte Colony Stimulating Factor (G-CSF); Granulocyte Macrophage Colony Stimulating Factor (GM-CSF); Granulocyte Chemotactic Proteins (GCPs); Growth Hormone (GH); I- 309; Growth Related Oncogene (GRO); lnhibins (Inh); Interferon-inducible T-cell Alpha Chemoattractant (I-TAC); Fas Ligand (FasL); Heregulins (HRGs); Heparin- Binding Epidermal Growth Factor-Like Growth Factor (HB-EGF); fms-like Tyrosine Kinase 3 Ligand (Flt-3L); Hemofiltrate CC Chemokines (HCC-1 to 4); Hepatocyte Growth Factor (HGF); Insulin; Insulin-like Growth Factors (IGF 1 and 2); Interferon- gamma Inducible Protein 10 (IP-10); lnterleukins (IL 1 to 18); Interferon-gamma (IFN-gamma); Keratinocyte Growth Factor (KGF); Keratinocyte Growth Factor-2 (FGF-10); Leptin (OB); Leukemia Inhibitory Factor (LIF); Lymphotoxin Beta (LT-B); Lymphotactin (LTN); Macrophage-Colony Stimulating Factor (M-CSF); Macrophage-derived Chemokine (MDC); Macrophage Stimulating Protein (MSP); Macrophage Inflammatory Proteins (MIPs); Midkine (MK); Monocyte Chemoattractant Proteins (MCP-1 to 4); Monokine Induced by IFN-gamma (MIG); MSX 1 ; MSX 2; Mullerian Inhibiting Substance (MIS); Myeloid Progenitor Inhibitory Factor 1 (MPIF-1 ); Nerve Growth Factor (NGF); Neurotrophins (NTs); Neutrophil Activating Peptide 2 (NAP-2); Oncostatin M (OSM); Osteocalcin; OP-1 ; Osteopontin; OX40 Ligand; Platelet derived Growth Factors (PDGF aa, ab and bb); Platelet Factor 4 (PF4); Pleiotrophin (PTN); Pulmonary and Activation-regulated Chemokine (PARC); Regulated on Activation, Normal T-cell Expressed and Secreted (RANTES); Sensory and Motor Neuron-derived Factor (SMDF); Small Inducible Cytokine Subfamily A Member 26 (SCYA26); Stem Cell Factor (SCF); Stromal Cell Derived Factor 1 (SDF-1 ); Thymus and Activation-regulated Chemokine (TARC); Thymus Expressed Chemokine (TECK); TNF and ApoL- related Leukocyte-expressed Ligand-1 (TALL-1); TNF-related Apoptosis Inducing Ligand (TRAIL); TNF-related Activation Induced Cytokine (TRANCE); Lymphotoxin Inducible Expression and Competes with HSV Glycoprotein D for HVEM T- lymphocyte receptor (LIGHT); Placenta Growth Factor (PIGF); Thrombopoietin (Tpo); Transforming Growth Factors (TGF alpha, TGF beta 1 , TGF beta 2); Tumor Necrosis Factors (TNF alpha and beta); Vascular Endothelial Growth Factors (VEGF-A₁B₁C and D).

2'-Deoxy-Nucleic acids (DNA):

[0146] DNA encodes the genes for proteins and peptides. Also, DNA contains a wide array of sequences that regulate the expression of the contained genes. Several types of DNA exist, depending on source, function, origin, and structure. Typical examples for DNA based molecules that can be utilized as bioactive, slow release coatings on probe tips (local gene-therapy) include, but are not limited to: A-DNA; B-DNA; artificial chromosomes carrying mammalian DNA (YACs); chromosomal DNA; circular DNA; cosmids carrying mammalian DNA; DNA; Double-stranded DNA (dsDNA); genomic DNA; hemi-methylated DNA; linear DNA; mammalian cDNA (complimentary DNA; DNA copy of RNA); mammalian DNA; methylated DNA; mitochondrial DNA; phages carrying mammalian DNA; phagemids carrying mammalian DNA; plasmids carrying mammalian DNA; plastids carrying mammalian DNA; recombinant DNA; restriction fragments of mammalian DNA; retroposons carrying mammalian DNA; single-stranded DNA (ssDNA); transposons carrying mammalian DNA; T-DNA; viruses carrying mammalian DNA; Z-DNA.

Ribo-Nucleic Acids (RNAs):

[0147] RNA is a transcription of DNA-encoded information. In some viruses, RNA is the essential information-encoding unit. Besides being an intermediate for expression of genes, RNAs have been shown to have several biological functions. Ribozymes are simple RNA molecules with a catalytic action. These RNAs can catalyze DNA and RNA cleavage and ligation, hydrolyze peptides, and are the core of the translation of RNA into peptides (the ribosome is a ribozyme).

[0148] Typical examples of RNA molecules with potential for use as bioactive coating on metal hybrid coated probe tips include, but are not limited to: acetylated transfer RNA (activated tRNA, charged tRNA); circular RNA; linear RNA; mammalian heterogeneous nuclear RNA (hnRNA), mammalian messenger RNA (mRNA); mammalian RNA; mammalian ribosomal RNA (rRNA); mammalian transport RNA (tRNA); mRNA; poly-adenylated RNA; ribosomal RNA (rRNA); recombinant RNA; retroposons carrying mammalian RNA ; ribozymes; transport RNA (tRNA); viruses carrying mammalian RNA.

Receptors:

[0149] Receptors are cell surface biomolecules that bind signals (e.g. hormone ligands and growth factors) and transmit the signal over the cell membrane and into the internal machinery of cells. Different receptors are differently "wired" imposing different intracellular responses even to the same ligand. This makes it possible for the cells to react differentially to external signals by varying the pattern of receptors on their surface.

[0150] Receptors typically bind their ligand in a reversible manner, making them suitable as carriers of growth factors that are to be released into the tissue. Thus by coating cantilever with growth factor receptors, and then load these receptors with their principal ligands, a bioactive surface is achieved that can be used for controlled release of growth factors to the surrounding tissues following implantation. Examples of suitable receptors with potential for use as bioactive coating on metal hybrid coated cantilever includes: The CD class of receptors CD; EGF receptors; FGF receptors; Fibronectin receptor (VLA-5); Growth Factor receptor, IGF Binding Proteins (IGFBP 1 to 4); lntegrins (including VLA 1-4); Laminin receptor; PDGF receptors; Transforming Growth Factor alpha and beta receptors; BMP receptors; Fas; Vascular Endothelial Growth Factor receptor (Flt-1 ); Vitronectin receptor.

Synthetic Biomolecules

[0151] Synthetic biomolecules are molecules that are based on and/or mimic naturally occurring biomolecules. By synthesizing such molecules a wide array of chemical and structural modification can be introduced that can stabilize the molecule or make it more bioactive or specific. Thus if a molecule is either too unstable or unspecific to be used from extracts it is possible to engineer them and synthesize them for use as implant surface coatings.

[0152] Furthermore, many biomolecules are so low abundant that extraction in industrial scales is impossible. Such rare biomolecules have to be prepared synthetically, e.g. by recombinant technology or by (bio-) chemistry. Below is listed several classes of synthetic molecules that can be potentially useful for implant coatings:

Synthetic DNA: [0153] Synthetic DNA molecules are biomolecules. These include, but are not limited to: A-DNA; antisense DNA; B-DNA; complimentary DNA (cDNA); chemically modified DNA; chemically stabilized DNA; DNA ; DNA analogues ; DNA oligomers; DNA polymers; DNA-RNA hybrids; double-stranded DNA (dsDNA); hemi-methylated DNA; methylated DNA; single-stranded DNA (ssDNA); recombinant DNA; triplex DNA; T-DNA; Z-DNA.

Synthetic RNA:

[0154] Synthetic RNA molecules are biomolecules. These include, but are not limited to: antisense RNA; chemically modified RNA; chemically stabilized RNA; heterogeneous nuclear RNA (hnRNA); messenger RNA (mRNA); ribozymes; RNA; RNA analogues; RNA-DNA hybrids; RNA oligomers; RNA polymers; ribosomal RNA (rRNA); and transport RNA (tRNA).

Synthetic Biopolymers:

[0155] The term biomolecules encompasses synthetic biopolymers, including but not limited to cationic and anionic liposomes; cellulose acetate; hyaluronic acid; polylactic acid; polyglycol alginate; polyglycolic acid; poly-prolines; polysaccharides.

Synthetic peptides:

[0156] Synthetic peptides are also encompassed by the term biomolecules. These peptides include, but are in no way limited to: decapeptides containing DOPA and/or diDOPA; peptides with sequence "Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys"; peptides where Pro is substituted with hydroxyproline; peptides where one or more Pro is substituted with DOPA; peptides where one or more Pro is substituted with diDOPA; peptides where one or more Tyr is substituted with DOPA; peptide hormones; peptide sequences based on the above listed extracted proteins; peptides containing an RGD (Arg GIy Asp) motif. Recombinant proteins:

All recombinantly prepared peptides and proteins

Synthetic Enzyme inhibitors:

[0157] Synthetic enzyme inhibitors range from simple molecules, like certain metal ions, that block enzyme activity by binding directly to the enzyme, to synthetic molecules that mimic the natural substrate of an enzyme and thus compete with the principle substrate. An implant coating including enzyme inhibitors could help stabilizing and counteract breakdown of other biomolecules present in the coating, so that more reaction time and/or higher concentration of the bioactive compound is achieved. Examples of enzyme inhibitors include, but are not limited to: pepstatin; poly-prolines; D-sugars; D-aminocaids; Cyanide; Diisopropyl fluorophosphates (DFP); metal ions; N-tosyl-l-phenylalaninechloromethyl ketone (TPCK); Physostigmine; Parathion; Penicillin.

Vitamins (synthetic or extracted):

[0158] The term biomolecules also encompasses synthetic and extracted vitamins, including but not limited to: biotin; calciferol (Vitamin D's; vital for bone mineralisation); citrin; folic acid; niacin; nicotinamide; nicotinamide adenine dinucleotide (NAD, NAD+); nicotinamide adenine dinucleotide phosphate (NADP₁ NADPH); NAD+retinoic acid (vitamin A); riboflavin; vitamin B's; vitamin C (vital for collagen synthesis); vitamin E; and vitamin K's.

Other bioactive molecules for AFM probe tip coatings:

[0159] Other suitable molecules for coating include, but are not limited to: adenosine di-phosphate (ADP); adenosine mono-phosphate (AMP); adenosine triphosphate (ATP); amino acids; cyclic AMP (cAMP); 3,4-dihydroxyphenylalanine (DOPA); 5'-di(dihydroxyphenyl-L-alanine (diDOPA); diDOPA quinone; DOPA-like o- diphenols; fatty acids; glucose; hydroxyproline; nucleosides; nucleotides (RNA and DNA bases); prostaglandin; sugars; sphingosine 1 -phosphate; and rapamycin.

Drugs for AFM probe tip coatings:

[0160] Pharmaceuticals incorporated in a hybrid layer of the probe tip coating may be utilized for local effects like improving local resistance against invading microbes, local pain control, local inhibition of prostaglandin synthesis; local inflammation regulation, local induction of biomineralisation and local stimulation of tissue growth. Examples of pharmaceuticals suitable for incorporation into metal hydride layers include, but are not limited to: Antibiotics; cyclooxygenase inhibitors; hormones; inflammation inhibitors; NSAID's; painkillers; prostaglandin synthesis inhibitors; steroids, tetracycline (also as biomineralizing agent).

Biologically active ions for AFM probe tip coatings:

[0161] Ions are important in a diversity of biological mechanisms. By incorporating biologically active ions in metal hybrid coated layers on cantilever it is possible to locally stimulate biological processes like enzyme function, enzyme blocking, cellular uptake of biomolecules, homing of specific cells, biomineralization, apoptosis, cellular secretion of biomolecules, cellular metabolism and cellular defense. Examples of bioactive ions for incorporation into metal hybrid coated include, but are not limited to: calcium; chromium; copper, fluoride; gold; iodide; iron; potassium; magnesium; manganese; selenium; silver; sodium; zinc.

[0162] In one embodiment of the invention, ALD may be used to form films containing biomolecules. In another embodiment, TMA (AI(CH₃)₃) molecules or similar precursors are pulsed into the chamber to form an initial reactive layer on the surface. A biological precursor is then pulsed into the chamber, forming bonds between the biomolecules and the aluminum atoms on the surface.

[0163] In another embodiment, the biological precursor may be serotonin. TMA (AI(CH₃)₃) molecules or similar precursors are pulsed into the chamber to form an initial reactive layer on the surface. Either the free amine group or the hydroxyl group will react with the aluminum atoms at the surface to form a film.

serotonin

[0164] Serotonin is a monoamine neurotransmitter synthesized in serotonergic neurons in the central nervous system (CNS) and enterochromaffin cells in the gastrointestinal tract of animals including humans. In the central nervous system, serotonin is believed to play an important role in the regulation of anger, aggression, body temperature, mood, sleep, vomiting, sexuality, and appetite. The bioamine serotonin (5-hydroxytryptamine; 5-HT), is a well-known neurotransmitter in the central nervous system. In addition, serotonin plays important roles in normal embryogenesis and cell growth, as well as being a regulator of physiological functions such as peristalsis in the gastrointestinal tract and blood pressure regulation. See Frishman et al., J Clin. Pharmacol., 35: 541- 572 (1995); Gershon et al., Aliment Pharmacol. Then, 13(Suppl 2): 15-30 (1999). In 2001 , a relationship between serotonin and bone was suggested by the demonstration of functional receptors for serotonin in osteoblastic cells. Westbroek et al., J. Cell. Biochem., 101 (2): 360-8 (2001 ). Serotonin may also regulate bone cell proliferation, differentiation, and activation in vitro. Bliziotes et al., Bone 29: 477-486 (2001 ); Westbroek et al., J. Cell. Biochem.,101 (2): 360-8 (2001 ); Gustafsson et al., J. Cell. Biochem. 97: 1283-1291 (2006). Moreover, long-term serotonin administration may result in increased bone mineral density (BMD), stiffer bones, and altered bone architecture in rats. Gustafsson et al., J. Cell. Biochem. 97: 1283-1291 (2006). These in vivo serotonergic effects on bone may be direct via serotonin receptors, but may also be indirect via interaction with other bone regulating substances such as leptin and adiponectin. J. Yamada et al., Eur. J. Pharmacol., 383: 49-51 (1999); J. Yamakawa et al., Diabetes Care, 26: 2477-2478 (2003). [0165] Serotonin and its transporter may also play a role in bone metabolism. The expression of the rate-limiting enzyme in serotonin synthesis, tryptophan hydroxylase, in osteoblasts and osteoclasts, has been demonstrated, indicating that bone cells may be capable of synthesizing serotonin. Gustafsson et al., J. Cell. Biochem., 98: 139-151 (2006). The membrane-bound serotonin transporter (5-HTT) expression has also been demonstrated in both osteoblasts and osteoclasts. Bliziotes et al., Bone 29: 477-486; Gustafsson et al., J. Cell. Biochem., 98: 139-151 (2006), and is responsible for the cellular internalization of serotonin, and is thus a key protein in serotonergic signaling and serotonin metabolism. The serotonin receptor-bearing bone cells may not only be able to respond to serotonin, but may also be able to regulate serotonin availability themselves, via its transporters as well as via synthesis.

[0166] Modulation of the serotonergic system may influence bone metabolism. In addition, long-term use of fluoxetine may effect on bone health. The exact mechanisms of serotonin action on bone metabolism are still unclear. Serotonin may act directly on the receptors on the bone cells, or may also act indirectly via other factors important in bone metabolism, like leptin and adiponectin. See Ducy et al., Cell 100: 197-207 (2000); Takeda et al., Cell, 111 : 305-317 (2002); Gordeladze and Reseland, J. Cell. Biochem., 88: 706-712 (2003); Luo et al., Exp Cell Res 309: 99-109 (2005); Oshima et al., Biochem. Biophys. Res. Commun., 331 : 520-526 (2005). In one embodiment of the invention, the use of serotonin coated probe tips may enhance the understanding of the relationship between serotonin and bone cells.

[0167] In another embodiment, ALD may be used to deposit biomolecular films on an AFM tip. These tips may optionally be used to image sample surfaces and to study the interactions between the bone-like material and sample surfaces.

[0168] Where a precursor contains two or more functional groups, the functional groups may be the same or different. The precursor should leave a reactive site suitable for succeeding growth with another type of precursor, unless no further deposition steps are desired. For production of the final terminating surface, the requirement of two or more types of functional groups does not apply. See WO 2006/071126A1. Rather the precursor should have at least one functional group that will undergo reactions with the previous surface, and also contain the groups that should finally terminate the surface.

[0169] Of particular interest is the study of interactions between biological materials and different types of surfaces. In one embodiment, probe tips may be coated with specific peptides using a sequence of amino acid analogs as ALD precursors. Peptides are formed by combining amino acids through amide bonds. For example, di-acyl chlorides of amino acids and amino acids with two amino groups may be used to deposit peptide films. Alternatively, dicarboxylic acids and diamines may be used to form peptide films.

[0170] These tips may then be used to probe biological surfaces. In one embodiment, depending on the sequence of the peptide, it will be possible to map the distribution, affinity, and dynamics of different receptors, for example, integrin receptors in a bone cell membrane. Magnetic/Conductive Coatings

[0171] Magnetic films may be constructed from magnetic materials that include, by way of non-limiting example, ferromagnetic, ferrimagnetic and paramagnetic materials. Examples of conductive films include, but are not limtied to ruthenium, palladium, molybdenum, TiN, LaNiθ₃, ZnO:AI, iridium, platinum and copper. Currently available MFM tips usually have a film thickness >30nm. One way to improve resolution of MFM is to reduce the film thickness, although this may lead to a reduction in the conductivity of the probe.

[0172] In at least one embodiment of the present disclosure, ALD may be used to deposit magnetic/conductive films on probe tips, for example Fe-Co with a thickness of 25±5 nm while maintaining stiffness of the probe equal to currently available MFM tips, thereby reducing film thickness without altering stiffness.

[0173] In one embodiment, a film is constructed by the ALD technique using suitable precursors for forming magnetic materials, according to the procedure described in Dalton Transactions, 2008, pgs. 253-259 by Lie et al., where Co(thd)₂ and Fe(thd)₃ in combination with ozone are used as the precursors to form (Fe₁Co)₃O₄ materials. The magnetic properties of the film may be further improved by annealing in a controlled oxygen and temperature environment.

[0174] In yet another embodiment, ALD may be used to deposit magnetic films by depositing an oxide or mixture of oxides that become ferro/ferri-magnetic when being subjected to suitable reduction or oxidation processes. A non-limiting list of examples are: Films of Fe₂O₃ may become ferrimagnetic Fe₃O₄ (magnetite) upon reduction, Cr₂O₃ may become ferromagnetic CrO₂ upon oxidation and the magnetic properties of manganite perovskites may be tuned depending on the oxidation potensial. In addition films containtn mixtures of oxides may be reduced to magnetic metallic states by suitable reduction processes such as annealing in pure H₂ at elevated temperatures, such as the process described in International Application No. WO 2002/045167. Hard Coatings

[0175] Hard coatings may be useful for maintaining sharp tips over time, for example when a physical contact tip-hard surface is used. In at least one embodiment, ALD may be used to deposit coatings on probe tips, for example to prolong the lifetime of the tip and the recording time of the probe. Thin layers of hard coatings enable conservation of the low spring constant of the probe and the geometry of the cantilever to remain unchanged over time.

[0176] Non-limiting examples of hard coatings include TiO₂, AI₂O₃, ZrO₂, Ti- Nitride, multi-layered materials and hybrid hard coatings.

[0177] In one embodiment, ALD may be used to coat SPM tips with hard coatings, for example by pulsing TiCI₄ and ammonia (NH₃) precursors into the reaction chamber onto the surface according to the ALD principle, using a carrier gas at elevated temperature and repeating until desired thickness is obtained for Ti- Nitride films, as described for example, in International Application No. WO 2007/013924 and Journal of Applied Physics, 2005, 97, 121301 , by Puurunen.

TiO? Coated SPM Probes

[0178] The stiffness of the cantilever of the probe and the radius of the probe tip in contact with the substrate are two parameters that may affect the resolution and the quality of the data.

[0179] The material composing the outer surface of the current commercially available probes include: SiN, Si₃N₄, and SiN coated with Au, Al or Pt, which may exhibit short life-times under certain conditions. Accordingly, the need to replace worn probes over short periods of time may be necessary, particularly for industries that use SPM applications requiring physical contact with the surface. [0180] Disclosed herein is a probe coated with Tiθ2, that may further strengthen the probe as compared to currently available silicon-based probes, and thus prolong the life-time of the SPM probes.

Example 1 Titanium Oxide AFM Tips

[0181] TiCI₄ and H2O were used to coat AFM tips with TiO₂. Films were grown in a commercial F-120 Sat reactor (ASM Microchemistry) by using TiCl₄ (Fluka; 98%) and H₂O (distilled) as precursors. Both precursors were kept at room temperature in vesels outside the reactor during the deposition. The reactor pressure was maintained at ca. 1.8 mbar by employing an N₂ carrier-gas flow of 300 cm³ min^~1 supplied from a Nitrox 3001 nitrogen purifier with a purity of 99.9995% inert gas (N₂ + Ar) according to specifications.

[0182] The films were grown using a pulsing scheme of 2 s pulse of TiCI₄ followed by a purge of 1 s. Water was then admitted using a pulse of 2 s followed by a purge of 1 s. This complete pulsing scheme makes up one pulsing cycle and the films were made using different numbers of such cycles (typically from 20-2000 cycles). Films can be formed in a relatively large temperature interval as shown in Figure 4. Using a deposition temperature of 150 ⁰C we obtained a growth rate of 0.054 nm/cycle.

[0183] The deposition may be expressed accordingly: Step 1 : TiCU(g) + I-OH → 1-0-TiCI₃ + HCI(g)

Step 2:

1-0-TiCI₃ + H₂O(g) → |-O-Ti-(OH)₃ + 3HCI(g)

[0184] The reactions may be shifted so that the liberation of HCI(g) is more in step 1 and less in step 2 depending on the reaction conditions. See R.L. Puurunen, J. Appl. Phys. 97 (2005) 121301.

[0185] By performing the deposition at a reactor temperature at or below 165 ⁰C, the resulting layer may be practically amorphous. The amorphous film may optionally be converted into the TiO₂ forms rutile or anatase by post annealing. Alternatively, the structure may be controlled in situ as described in J. Aarik et al., J. Cryst. Growth 148: 268 (1995) where anatase is deposited in the range 165 - 350 ⁰C and rutile is obtained at temperatures above 350 ⁰C.

[0186] Alternatively, polycrystalline films of pure rutile, pure Tiθ₂-ll, or a mixture of them both may be formed. See J. Aarik, A. Aidla, V. Sammelselg, H. Siimon, T. Uustare, J. Cryst. Crowth 169 (1996) 496. Polycrystalline films of pure rutile, pure TiO₂-II, or a mixture of them both can be produced by varying the water pressure during deposition at 400 ⁰C for otherwise similar conditions using TiCI₄ and H₂O as precursors. The TiO2-ll structure is an orthorhombic phase isomorphous to D-PbO₂.

[0187] Molecular force probing (MFP/AFM Asylum research, Santa Barbara U.S.) with titanium oxide coated AFM cantilevers (AC160 Olympus SilicaNitril, Tokyo, Japan) with tip radii 10 nm and height 10 μm was used to scan mouse pre- osteoblasts (donated) in DMEM (Dulbecco/Vogt Modified Eagle's Minimal Essential Medium, Cambrex Biosciences, UK), at 22 ⁰C in aqueous contact mode.

[0188] Mouse pre-osteoblasts (Type, MC3T3 E1 , CRL-2593, ATCC, Manassas, USA) were placed with DMEM in an environmental chamber (Bioheater, Asylum Research, USA) on a polymer slide (NUNC™ Brand Pocket, SlideFlask, Roskilde, Denmark) without any fixation chemicals.

[0189] 400 force curves were generated. Representative curves are shown in Figures 5 and 6. Figure 5 reveals strong adhesion between the TiO₂-coated AFM tip and the osteoblast.

[0190] These force curves were then used to generate a map of adhesion strengths across the cell surface, shown in Figure 7.

Example 2 Titanium Oxide Coated SPM Cantilevers

[0191] SPM cantilevers (RC-800PSA, Olympus, Tokyo, Japan) were coated in a commercial F-120 Sat reactor (ASM Microchemistry) by using TiCI₄ (Aldrich; 99%) and H₂O (distilled) as precursors, such as the process described in J. Aarik, A. Aidla, E. Uustare, V. Sammelselg, Journal of Crystal Growth, 148(1995) 268. Both precursors were kept at room temperature in vessels outside the reactor during the deposition and pulsed into the reactor without an external carrier gas. [0192] To provide saturated growth, the deposition process followed a pulsing scheme of 0.6 s pulse of TiCI₄ followed by a 1 s purge. Water was then admitted using a 2 s pulse followed by a 1 s purge. Deposition was performed at three different reactor temperatures: 150⁰C, 300⁰C, and 400⁰C conducive to providing amorphous, anatase and rutile phases respectively. Using deposition temperatures of 150°C and 300°C, we obtained a growth rate of 0.046 and 0.040 nm/cycle respectively.

[0193] A silicon substrate was subjected to the deposition process described above and compared with cantilevers coated with titanium oxide to measure and compare film thickness and crystallinity.

[0194] The crystallinity of the films was analyzed by x-ray diffraction (Siemens D5000 diffractometer) in Θ-2Θ mode using CuKa radiation. The results from the x-ray diffraction analysis are shown in Figure 16. SEM was used to measure TiO₂ film thickness using a TM-1000 (Hitachi tabletope microscope, Tokyo, Japan) and a Philips XL 30 ESEM (FEI Electronics Optics, Eindhoven, Netherlands). The results for these measurements are shown in Figures 17 and 18.

[0195] Cantilever stiffness, according to coating thickness, was also analyzed by AFM. AFM (MFP-3D, Asylum research, Santa Barbera, U.S.) was performed on uncoated cantilevers and TiO₂-coated cantilevers with seven different thicknesses: 1 , 5.4, 6, 10.9, 12.5, 30, 50, 75, 98.4, 114, and >120nm. Spring constant (i.e. stiffness of the cantilevers) was measured by force curve analysis. The results are shown in Figure 19.

[0196] MFP was used to analyzed adhesion forces between coated and non-coated cantilevers versus cell culture plates. 900 force-curves were performed using MFP to test the physical/chemical activation of the tip surface by a 6nm TiO₂ coating compared with SiN non-coated tip. The substrates used for analysis were the Nunclon™Δ Surface (NUNC A/S, Roskilde, Denmark), and collagen I coated plates (NUNC A/S, Roskilde, Denmark). The tests were performed in deionized water at room temperature. Results are shown in Figures 20 and 21. Example 3 Ti-O-N Surfaces

[0197] TiO_xNy surfaces may be produced by varying the usage of H₂O or NH₃ as precursor in the reaction scheme described for growth of TiO₂. The reaction scheme may be as follows: Step 1 : TiCU(g) + I-OH → 1-0-TiCI₃ + HCI(g)

Step 2a:

1-0-TiCI₃ + 3H₂O(g) → |-O-Ti-(OH)₃ + 3HCI(g)

Step 2b:

1-0-TiCI₃ + 3NH₃(g) → |-O-Ti-(NH₂)₃ + 3HCI(g)

Example 4 Calcium Carbonate

[0198] Calcium carbonate films were grown in a commercial F-120 Sat reactor (ASM Microchemsitry) by using Ca(thd)₂ (Strem Chemicals; 99.9%; Hthd = 2,2,6,6-tetramethylheptan-3,5-dione) and ozone as precursors, with CO₂ (AGA; 99.99 %) as controlling atmosphere in selected runs. Ozone gas with a flow of 500 cm³ min^"1 was produced by feeding pure O₂ (AGA 99.999%) into an ozone generator (OT-020, OzoneTechnology; giving an ozone concentration of 15 vol.% according to specifications). The applied sublimation temperature for Ca(thd)₂ was 195 ⁰C. The reactor pressure was maintained at ca. 1.8 mbar by employing an N₂ carrier-gas flow of 300 cm³ min^"1, supplied from a Nitrox 3001 nitrogen purifier with a purity of 99.9995% inert gas (N₂ + Ar) according to specifications. Additional CO₂ (AGA 99%) was introduced to produce well crystalline carbonate. The CO₂ was introduced at a flow of approximately 400 cm³ min^"1.

[0199] The films were made using a pulsing scheme of 1 s Ca(thd)₂ followed by a purge of 0.5 s and then a 2 s pulse of O₃ followed by a purge of 0.8 s. In addition to this sequence, an additional pulse of 3 s CO₂ was introduced followed by a purge of 1 s. This sequence forms a cycle may be repeated until the desired film thickness is achieved. Films using a total of 250 - 5000 cycles have been produced. [0200] Under these conditions, the growth rate varied relatively little with deposition temperature, and was at ca. 0.045 nm/ cycle, as shown in Figure 8.

Example 5 Calcium Phosphates

[0201] Calcium phosphate films may be grown in a commercial F-120 Sat reactor (ASM Microchemsitry) by using Ca(Cp)₂ (Cp = cyclopentadienyl), H₂O (distilled), and POCI₃ (99% Fluka) as precursors. Ca(Cp)₂ may be sublimed at ca. 85 °C, and H₂O and POCI₃ may be kept at room temperature in vessels outside the reactor during the deposition. The reactor pressure may be maintained at ca. 1.8 mbar by employing an N₂ carrier-gas flow of 300 cm³ min^~1, supplied from a Nitrox 3001 nitrogen purifier with a purity of 99.9995% inert gas (N₂ + Ar) according to specifications.

[0202] The films may be grown using a pulsing scheme of 2 s pulse of Ca(Cp)₂ followed by a purge of 1 s. Water may then be admitted using a pulse of 2 s followed by a purge of 1 s. POCI₃ may then be pulsed for 2 s followed by a purge of 1 s. Water may then be admitted using a pulse of 2 s followed by a purge of 1 s. This pulsing scheme comprises one pulsing cycle. Films may be made using different numbers of these cycles (typically from 20-2000 cycles). Films may be formed at suitable deposition temperatures. The films may be made by repeating the pulsing cycles until the desired thickness was achieved.

[0203] The deposition may be expressed as follows: Step 1 : I-OH + Ca(Cp)₂(g) = |-O-Ca-Cp + H-Cp(g)

Step 2:

I-Ca-Cp + H₂O(g) = |-Ca-OH + H-Cp(g)

Step 3:

2I-OH + POCI₃(g) = |-PO₃CI + 2HCI(g)

Step 4:

1-PO₃CI + H₂O(g) = 1-PO₄H + HCI(g) [0204] Alternatively, films may be grown as follows. Films may be grown using Ca(Cp)2 (Cp = cyclopentadienyl), H₂O (distilled), and PPh₃ (Ph = phenyl) (98.5% Fluka) as precursors. Ca(Cp)₂ may be sublimed at ca. 85 ⁰C, while H₂O and PPh₃ may be kept at room temperature in vessels outside the reactor during the deposition. The reactor pressure may be maintained at ca. 1.8 mbar by employing an N₂ carrier-gas flow of 300 cm³ min^~1, supplied from a Nitrox 3001 nitrogen purifier with a purity of 99.9995% inert gas (N₂ + Ar) according to specifications.

[0205] The films may grown using a pulsing scheme of 2 s pulse of Ca(Cp)₂ followed by a purge of 1 s. Water may then be admitted using a pulse of 2 s followed by a purge of 1 s. PPh₃ may then be pulsed for 2 s followed by a purge of 1 s. Water may then be admitted using a pulse of 2 s followed by a purge of 1 s. Ozone may was be then admitted for 3 s followed by a purge of 1 s. This complete pulsing scheme makes up one pulsing cycle and the films were made using different numbers of such cycles (typically from 20-2000 cycles). Films may be formed at suitable deposition temperatures. The films may be made repeating the pulsing cycles until the desired thickness is achieved.

[0206] This deposition sequence may be expressed as follows: Step 1 : I-OH + Ca(Cp)₂(g) = |-O-Ca-Cp + H-Cp(g)

Step 2:

I-Ca-Cp + H₂O(g) = |-Ca-OH + H-Cp(g)

Step 3:

I-OH + P(Ph)3(g) = |-O-P(Ph)2 + H-Ph(g)

Step 4:

1-0-P(Ph)₂ + 2H₂O(g) = |-O-P(OH)₂ + 2H-Ph(g)

Step 5: 1-0-P(OH)₂ + O₃(g) = 1-0-PO(OH)₂ + O2(g)

[0207] Alternatively, ozone may be pulsed in place of H₂O in step 4. Step 5 may then be expressed as: Step 4b: |-O-P(Ph)₂ + xO₃ = |-O-PO(OH)₂ + CO₂(g) + H₂O(g)

[0208] Alternatively, the process may be modified to produce fluorapatite by pulsing NH₄F in place of water in one or more pulses according to the reaction:

Ca₂(PO₄)(OH) + NH₄F = Ca₂(PO₄)F + NH₃ + H₂O

[0209] An alternative approach to obtain Ca-P-O films is to use the precursor pairs Ca(thd)₂ + O₃ and (CH₃O)₃PO + H₂O as precursors in a mixed fashion. The film is formed by first producing a monolayer of CaCO₃ using the Ca(thd)₂ + O₃ precursor pair as described in Example 3 and theafter transforming this into Ca₃(PO₄)₂ by using the (CH₃O)₃PO + H₂O precursor pair. The Ca₃(PO₄J₃ film can be transformed into hydroxyapatite by subsequent treatment of the film in moist N₂ at temperatures above 500 ⁰C.

Example 6 Calcium Fluoride

[0210] CaF₂ films may be produced using Ca(thd)₂ and HF as precursors. See M. Ylilammi & T. Ranta-aho, J. Electrochem. Soc, 141 : 1278 (1994).

[0211] The reactions may proceed as follows. Overall: Ca(thd)₂ + 2HF = CaF₂ + 2Hthd Step 1 : |-Ca-F₂ + Ca(thd)2(g) = |-Ca-F₂-Ca(thd)₂

Step 2: |-Ca-thd2 + 2HF(g) = |-Ca-F₂ + 2Hthd(g) [0212] Alternatively, the fluoride may be introduced using TiF₄ or TaF₅ as precursors. The films may be formed according to the reaction scheme:

2Ca(thd)₂(g) + TiF₄(g) = 2CaF₂(S) + Ti(thd)₄(g)

5Ca(thd)₂(g) + 2TaF₅(g) = 5CaF₂(S) + 2Ta(thd)₅(g)

[0213] The TiF₄ precursor may be sublimed in the reactor at 140-145 ⁰C. The TaF₅ precursor may be sublimed in the reactor at 45-50 ⁰C. The Ca(thd)2 precursor may be sublimed in the reactor at 195 °C. The pulse times for Ca(thd)₂ may be 2 s or longer followed by a purge of 1 s. The pulse time for TiF₄ or TaF₅ may be 1 s followed by a purge of 1 s.

Example 7 Glycine Hybrid Films

[0214] Glycine hybrid films were grown in a commercial F-120 Sat reactor (ASM Microchemsitry) by using either TiCI₄ (Fluka; 98%) or TMA (trimethylaluminium; Withco, 98%) and glycine (Aldrich, 99%). The metal precursors were kept at room temperature in vessels outside the reactor during the deposition. Glycine was sublimed in the reactor at 200 ⁰C. The reactor pressure was maintained at ca. 2 mbar by employing an N₂ carrier-gas flow of 300 cm³ min^~1 supplied from a Nitrox 3001 nitrogen purifier with a purity of 99.9995% inert gas (N₂ + Ar) according to specifications.

[0215] The films were grown using a pulsing scheme of 1 s pulse of TiCI₄ or TMA followed by a purge of 2 s. Glycine was then admitted using a pulse of 2 s followed by a purge of 2 s. This complete pulsing scheme makes up one pulsing cycle and the films were made using different numbers of such cycles (typically from 2-2000 cycles). Quarts crystal microbalance results from such measurements performed at 250 °C are shown in Figures 9 and 10. Typical growth rates of these films at 250 ⁰C are 0.47 nm/cycle for Ti-Glycine and 0.53 nm/cycle for Al-Glycine. Example 8 4-Aminobenzoic Acid Hybrid Films

[0216] 4-Aminobenzoic acid (Aldrich; 99%) films were grown in a commercial F-120 Sat reactor (ASM Microchemsitry) by using TiCI₄ (Fluka; 98%). The metal precursors were kept at room temperature in vessels outside the reactor during the deposition. 4-aminobenzoic acid was sublimed in the reactor at 160 °C. The reactor pressure was maintained at ca. 2 mbar by employing an N₂ carrier-gas flow of 300 cm³ min^~1, supplied from a Nitrox 3001 nitrogen purifier with a purity of 99.9995% inert gas (N₂ + Ar) according to specifications.

[0217] The films were grown using a pulsing scheme of 1 s pulse of TiCI₄ followed by a purge of 1 s. 4-aminobenzoic acid was then admitted using a pulse of 2.5 s followed by a purge of 1 s. This complete pulsing scheme makes up one pulsing cycle and the films were made using different numbers of such cycles (typically from 2-2000 cycles). Quarts crystal microbalance results from such measurements performed at 200 ⁰C are shown in Figure 11. Typical growth rates of the compound at 200 °C are: 1.0 nm/cycle for Ti-4-aminobenzoic acid.

Example 9 4-Aminobenzophenone Hybrid Films

[0218] 4-Aminobenzophenone hybrid films were grown in a commercial F- 120 Sat reactor (ASM Microchemsitry) by using TiCI₄ (Fluka; 98%). The metal precursors were kept at room temperature in vessels outside the reactor during the deposition. 4-aminobenzophenone (Fluka; 97%) was sublimed in the reactor at 126 °C. The reactor pressure was maintained at ca. 2 mbar by employing an N₂ carrier- gas flow of 300 cm³ min^"1, supplied from a Nitrox 3001 nitrogen purifier with a purity of 99.9995% inert gas (N₂ + Ar) according to specifications.

[0219] The films were grown using a pulsing scheme of 1 s pulse of TiCI₄ followed by a purge of 1 s. 4-aminobenzophenone was then admitted using a pulse of 2.5 s followed by a purge of 1 s. This pulsing scheme makes up one pulsing cycle, and the films were made using different numbers of such cycles (typically from 2-2000 cycles). Quarts crystal microbalance results from such measurements performed at 200 ⁰C are shown in the Figure 12. Example 10 Sertonin Hybrid Films

[0220] Sertonin films may be grown in a commercial F-120 Sat reactor (ASM Microchemsitry) by using TiCI₄ (Fluka; 98%). The metal precursors may be kept at room temperature in vessels outside the reactor during the deposition. Serotonin hydrochloride (Sigma) may be sublimed in the reactor at a suitable sublimation temperature, which may be approximately 200 °C. The reactor pressure may be maintained at ca. 2 mbar by employing an N₂ carrier-gas flow of 300 cm³ min^~1, which may be supplied from a Nitrox 3001 nitrogen purifier with a purity of 99.9995% inert gas (N₂ + Ar) according to specifications.

[0221] A suitable pulsing scheme may employ a 1 s pulse of TiCI₄ followed by a purge of 1 s. Serotonin may then be admitted using a 2.5 s pulse followed by a purge of 1 s. This example pulsing scheme makes up one pulsing cycle. Ti- sertonin films may be made using different numbers of such cycles (typically from 2-2000 cycles).

[0222] Serotonin has both a hydroxyl and two amines as functional groups and should react according to the reaction mechanisms proposed above for hybrid films.

Example 11 Amino acids and Peptide Films

[0223] Films of amino acids may be deposited by forming peptide bonds according to the reaction scheme shown below.

[0224] The carboxylic acids may be reacted with amines, forming water as a byproduct. See M. Putkonen et al., J. Mater. Chem., 17: 664 (2007).

[0225] Alternatively, the chloride salts of the carboxylic acids may be reacted with amines, forming HCI as a byproduct, as shown below. The iodine or bromine salts may also be used. See A. Kubono et al., Thin Solid Films 289: 107 (1996). Overall reaction:

CI-OC-R-CO-CI + H₂N-R'-NH₂ = [CI-OC-R-CO-NH-R'-NH₂] + HCI(g)

Step 1 :

I-R-CO-CI + H₂N-R'-NH₂(g) = |-R-CO-NH-R'-NH₂ + HCI(g)

Step 2:

|-R-CO-NH-R'-NH₂ + CI-OC-R-CO-CI(g) = I-R-CO-NH-R'-HN-OC-R-CO-CI + HCI(g)

[0226] Alternatively, molecules of the type: Y₁-NH-R-CO-CI + H₂N-R'-CO- Y₂ may be used to deposit films where each Y group may be a blocking agent that prevents self-polymerization. In one embodiment, Yi and/or Y₂ may be a substituted or unsubstituted alkyl group. Step 1 : I-R-CO-CI + H₂N-R^NH-Y₁Cg) = I-R-CO-NH-R'-NHΛ^ + HCI(g)

Step 2: l-R-CO-NH-R'-NH-Yi + H₂O(g) = |-R-CO-NH-R'-NH₂ + YrO-H(g)

Step 3:

|-R-CO-NH-R'-NH₂ + CI-OC-R-CO-Y₂(g) = |-R-CO-NH-R'-H N-OC-R-CO-Y₂ + HCI(g)

Step 4:

|-R-CO-NH-R'-HN-OC-R-CO-Y₂ + H₂O(g) = I-R-CO-NH-R'-HN-OC-R-CO-OH +

Y₂H(g)

[0227] Alternatively, molecules of the type: H₂N-R-C=C-R' or preferrably H₂N-R-C≡C-R' may be used together with ozone to form peptide bonds. The reaction mechanism may utlilize ozonolysis of the unsaturated bonds (preferrably the alkyne bond) to transform these into carboxylic acid or carboxylic anhydride groups. The reaction mechanism for formation of carboxylic anhydride groups from alkyne bonds is given in the figure below: R ≡Ξ≡Ξ R'

In,

[0228] Molecules of the type H₂N-R-C≡C-R' may be pulsed into a reaction chamber. The amine group may then react with the previous surface to form a peptide bond. This reaction may occur, but is not limited to, through an anhydride on that surface. Excess precursor may then optionally be removed from the reaction chamber. Ozone may then be admitted. The ozone may react with the alkyne bond to form a carboxylic anhydride. Excess ozone may then be removed from the reaction chamber. The resulting surface may then consist of carboxylic anhydrides that may function as reactive goups for formation of new peptide bonds by admission of further amines and liberation of a carboxylic acid containing the R' group. The process may thus be repeted with the same type or different types of amines. When molecules of the type H₂N-R-C=C-R' are used as precursors, the ozonolysis may result in carboxylic acid groups that may be used for further reaction with amines and/or amine salts to produce peptide bonds.

Example 12 Large Biomolecules

[0229] Biomolecular materials may be attached to surfaces as follows. A surface (such as an AFM tip) may be treated with or exposed to water vapour or liquid water to form a terminating hydroxyl layer.

[0230] In an inert atmosphere, this surface may then be dipped into a solution of TMA or other highly reactive metal complexes such as, but not limited to, TiCU, diethyl zinc, magnesium cyclopendadienyl, in an inert organic solvent, for example but not limited to, heptane, hexane, or toluene. The concentration of TMA (metal complex) may be in the range of, but not limited to, 0.0001 - 10 M in the inert solvent. This surface may then be rinsed gently with an inert organic solvent to remove excess TMA or other metal complex. [0231] The surface may then be dipped into a separate solution containing an organic biomolecule. This biomolecule will then attach to the surface by reaction with the reactive metal complexes. The organic biomolecules may be in an inert organic solvent. The identity of the biomolecule may be any biomolecule containing a functional group that will react with the methyl-aluminum (or other metal complex) surface. Suitable biomolecules include but are in no way limited to RGD peptide and RNA and/or DNA molecules.

Example 13

[0232] AFM tips coated with peptides can be used to map and probe receptors on cell membrane surfaces. This is illustrated in Figure 13. An AFM cantilever tip may be coated with peptide ("A"). Molecular force probing can then be used to map the surface of a bone cell. If A contains the motif Arg-Gly-Asp (RGD) tri-peptides, there will be an adhesive force ("B") between the peptide and the integrin receptor ("C") on the cell membrane. This adhesive force will be specific to the interaction between peptide A and the integrin (α_vβ₃/ α_vβ₅ and α₅βi). The peptide will not interact as strongly with other receptors, represented by "D" in Figure 13. Therefore, a map of force curves can reveal the location, distribution, and abundance of the receptor specific to the peptide on the AFM tip.

[0233] Depending on the peptide sequence, one can obtain topo-dynamic three-dimensional maps of different receptors on the cell membrane. By switching between tips coated with different materials, the distribution in 3D of several competing or interacting receptors on the same cell can be mapped and the binding forces between ligands and receptors measured.

Example 14

[0234] An AFM cantilever coated with serotonin according to the invention described herein (such as in Example 9) may be used in an atomic force microscope equipped for observation in an incubation chamber that is designed to facilitate the support of living human bone cells (such as the MFP-3D, Asylum research Santa Barbara, US). The cantilever may be brought in contact with the surface of a cell in the incubation chamber, and may be used to scan the cell surface while the microscope is in MFP mode (molecular force probing). The MFP mode may allow for recoding of adhesive forces between the cantilever and the cell surface. An affinity map representing the distribution of serotonin receptors may be constructed of the cell surface. The distribution, concentration, modulation, and dynamics of the cell's serotonin receptors may be analyzed while they are growing on different substrates, in different media, and at different time points in differentiation. In addition, pharmaceuticals (like SSRIs) and biological signal molecules (i.e. hormones, cytokines, fluoride, calcium, growth factors etc) may be introduced to the system while the cell is being observed to investigate the influence of such additives. In addition, the serotonin-coated AFM cantilever may be used to "load" (mechanically stress) single cells and at the same time may be used to monitor the serotonin receptor activity, thus analyzing the activity and function of the serotonin system in real time in a living loaded cell. To detect and measure adhesion force at around pico newton level, the cantilever used in the MFP mode may be elastic (with a spring constant less than 0.07 nN/nm). The adhesion forces is applied on living cells (e.g. human osteoblasts, or any other cell line, or human cell culture) placed in appropiate growth media (e.g. Osteoblast Basal Medium, OBM TM, Clonetics, Cambrex, Walkersville, US). The living cells and medium may be placed in an environmental chamber (Bioheater ®, Asylum Research, Santa Barbara, US) in order to keep temperature constant at 37 ⁰C or alter the temperature to any desired value. The mapping of the osteoblast's adhesion forces was conducted in an area of 80 μm x 80 μm, with 400 adhesion points within this area. The cantilever was moved down toward the cell at a speed of 500 nm/sec and retracted at the same speed. The deflection's trigger point for the cantilever when the serotonin coated cantilever tip touches the cell's membrane was 15 nm. The adhesion forces between the serotonin coated cantilever and the specific serotonin receptor on the osteoblast cell membrane, was higher regions without receptors. Example 15

[0235] The wear resistance of A) uncoated, commercially available (SiN) AFM cantilevers (control) were compared with AFM cantilevers coated with a thin layer (=10nm) of B) titanium dioxide (TiO₂), C) zirconium dioxide (ZrO₂) and D) aluminum oxide (AI2O₃). [0236] AFM cantilevers coated with «10nm of TiO₂ were prepared according to the present invention. A layer of TiO₂ was deposited using the ALD (atomic layer deposition) technique in a F-120 Sat reactor (ASM Microchemistry). The deposition was performed using TiCI₄ (Fluka) and H₂O (distilled) as precursors at a deposition temperature of 150 ⁰C. Both precursors were used at room temperature. A thickness of 10 nm was reached after 148 deposition cycles.

[0237] AFM cantilevers coated with =10nm of ZrO₂ were prepared according to the present invention. A layer of ZrO₂ was deposited using the ALD (atomic layer deposition) technique in a F-120 Sat reactor (ASM Microchemistry). The deposition was performed using ZrCI₄ (Aldrich) and H₂O (distilled) as precursors at a deposition temperature of 250 ⁰C. Both precursors were used at room temperature. A thickness of 10 nm was reached after 67 deposition cycles.

[0238] AFM cantilevers coated with =10nm of AI₂O₃ were prepared according to the present invention. A layer of AI₂O₃ was deposited using the ALD (atomic layer deposition) technique in a F-120 Sat reactor (ASM Microchemistry). The deposition was performed using AI(CH₃)₃ (trimethylaluminium, TMA) (Witco) and O₃ as precursors at a deposition temperature of 300 ⁰C. The TMA precursor was used at room temperature while the O₃ precursor was delivered from an OT- 020 ozone generator provided with 99.999% O₂ (AGA) at a rate of 500 seem. A thickness of 10 nm was reached after 91 deposition cycles.

[0239] All wear resistance tests were performed at room temperature (23⁰C), alternative contact mode (AC mode), using a MFP-3D AFM (Asylum Research, Santa Barbara, USA). The hardness and wear resistance of the cantilevers were tested against ZrO₂ surfaces. The Q was set at 110, in mixed mode (attractive and repulsive imaging). Q is a parameter that improves the phase contrast imaging (the shift from repulsion to attraction gets more sensitive with a high Q). Keeping a constant Q gives comparatives measurements possible between cantilevers. The free amplitude in AC mode was set around 60 nm, and the set point during scanning fixed at 40 nm. 18 scans were performed with the same cantilever, the same parameters, over the same area.

[0240] SEM (Philips XL 30 ESEM, FEI Electron Optics, Eindhoven, Netherlands) was also used to compare images of the cantilever tips tbefore and after AFM scanning. The before and after images were also superposed to reflect differences in wear damage of the tip due to scanning friction against the ZrO₂ surfaces between A) uncoated cantilevers, B) cantilevers coated with TiO₂, C) cantilevers coated with ZrO₂, and D) cantilevers coated with AI₂O₃. Results are shown in Figures 22-29. Example 16

[0241] Commercially available cantilevers (Olympus Si AC240, uncoated; ASYMFM, coated with » 50nm cobalt chrome; and Olympus RC800, uncoated) were compared with cantilevers coated with magnetic coatings according to the invention to determine magnetic field sensitivity.

Olympus Si AC240 and ASYMFM cantilevers

[0242] A commercially available Olympus Si AC240 cantilever and a commercially available MFM cantilever (ASYMFM) served as controls (i.e., no surface modification was performed).

[0243] An Olympus Si AC240 cantilever was coated with Fe₂CoO₄ (thickness « 10nm) according to the invention. A layer of (Co₁Fe)₃O₄ was deposited using the ALD (atomic layer deposition) technique in a F-120 Sat reactor (ASM Microchemistry). The deposition was performed using Co(thd)₂ and Fe(thd)₃ (Hthd = 2,2,6, 6-tetramethyl-3,5-heptadione) and O₃ as precursors at a deposition temperature of 200 ⁰C. The Fe(thd)₃ precursor was sublimed at 115 ⁰C, the Co(thd)₂ precursor was sublimed at 118 ⁰C while the O₃ precursor was delivered from an OT-020 ozone generator provided with 99.999% O₂ (AGA) at a rate of 500 seem. A thickness of ca. 10 nm was reached after 1030 deposition cycles of metal.

[0244] The magnetic experiments for all of the Olympus Si AC240 and ASYMFM cantilevers were performed at room temperature (23 ⁰C), in alternative contact mode (AC mode), using a MFP-3D atomic force microscope (AFM) (Asylum Research, Santa Barbara, USA) equipped with a variable field module (VFM, Asylum Research, Santa Barbara, USA). The ability of the cantilevers in detecting any magnetic field on a 3¹/4-inch floppy disk full with data was tested and used as a reference for the other tested surface coatings.

[0245] The Q was set at 200, in mixed mode (attractive and repulsive imaging). Q is a parameter that improves the phase contrast imaging (the shift from repulsion to attraction gets more sensitive with a high Q). Keeping a constant Q gives comparatives measurements possible between cantilevers. The free amplitude in AC mode was set around 60 nm, and the set point during scanning fixed at 45 nm. The magnetic scanning was performed at a distance of 100 nm above the analysed surface (floppy disk). The scan size was chosen at 30 μm x 30 μm and the scan speed at 75.12 μm/s. The target field of the VFM stage was kept close to 0 Gauss in order to keep the data on the floppy disk. (Increasing this Field to almost 1000 Gauss erases the magnetic storage of the disk). Results are shown in Figures 30-32.

Olympus RC800 cantilevers

[0246] A commerically available Olympus RC800 PSA cantilever, SiN (uncoated) served as control (i.e., no surface modification).

[0247] An Olympus RC800 PSA cantilever was coated with Fe₂O₃ (thickness « 10nm) according to the invention. A layer of Fβ₂θ₃ was deposited using the ALD (atomic layer deposition) technique in a F-120 Sat reactor (ASM Microchemistry). The deposition was performed using Fe(thd)₃ (Hthd = 2,2,6,6- tetramethyl-3,5-heptadione) and O₃ as precursors at a deposition temperature of 200 ⁰C. The Fe(thd)₃ precursor was sublimed at 115 ⁰C while the O₃ precursor was delivered from an OT-020 ozone generator provided with 99.999% O₂ (AGA) at a rate of 500 seem. A thickness of 10 nm was reached after ca. 834 deposition cycles.

[0248] An Olympus RC800 PSA cantilever was coated with Fe₂CoO₄ (thickness « 10nm) according to the invention. A layer of Fe₂CoO₄ was deposited using the ALD (atomic layer deposition) technique in a F-120 Sat reactor (ASM Microchemistry). The deposition was performed using Co(thd)₂ and Fe(thd)₃ (Hthd = 2,2,6, 6-tetramethyl-3,5-heptadione) and O₃ as precursors at a deposition temperature of 200 ⁰C. The Fe(thd)₃ precursor was sublimed at 115 ⁰C, the Co(thd)₂ precursor was sublimed at 118 ⁰C while the O₃ precursor was delivered from an OT-020 ozone generator provided with 99.999% O₂ (AGA) at a rate of 500 seem. A thickness of ca. 10 nm was reached after 1030 deposition cycles of metal.

[0249] The magnetic experiments with the Olympus RC800 cantilevers were performed at room temperature (23 ⁰C), in contact mode, using a MFP-3D atomic force microscope (AFM) (Asylum Research, Santa Barbara, USA) equipped with a variable field module (VFM, Asylum Research, Santa Barbara, USA). The ability of the cantilevers in detecting any magnetic field on a 3¹/2-inch floppy disk full with data was tested and used as a reference for the other tested surface coatings. [0250] The magnetic scanning was performed at a distance of 300 nm above the analysed surface (floppy disk). The scan size was chosen at 30 μm x 30 μm and the scan speed at 75.12 μm/s. The target field of the VFM stage was kept close to 0 Gauss in order to keep the data on the floppy disk. (Increasing this Field to almost 1000 Gauss erases the magnetic storage of the disk). Results are shown in Figures 33-35.

Claims

WHAT IS CLAIMED IS:

1. A functionalized AFM probe comprising an AFM tip coated with at least one material chosen from biological, organic, organic-inorganic hybrid, bone- like, and an implant-like material.

2. A functionalized AFM probe of claim 1 , wherein the probe comprises at least one material chosen from Si, Si₃N₄, and SiN.

3. A functionalized AFM probe of claims 1 -2, wherein the probe further comprises gold, aluminum, or platinum.

4. A functionalized AFM tip of claims 1-3, wherein the biological material comprises a hybrid film comprising one or more biomolecules chosen from neurotransmitters, ligands, amino acids, antibodies, sugars, peptides, proteins, fatty acids, spingolipids, and lipids.

5. A functionalized AFM tip of claims 1-4, wherein the biological material comprises a hybrid film comprising one or more biomolecules chosen from bioadhesives, cell attachment factors, biopolymers, blood proteins, enzymes, extracellular matrix proteins and biomolecules, growth factors and hormones, 2'- deoxy-nucleic acids, ribo-nucleic acids, receptors, synthetic biomolecules, synthetic DNA, synthetic RNA, synthetic biopolymers, synthetic peptides, recombinant proteins, synthetic enzyme inhibitors, synthetic and extracted vitamins, pharmaceuticals, and biologically active ions.

6. A functionalized AFM tip of claim 5, wherein one or more bioadhesive is chosen from marine mussel adhesive proteins, fibrin-like proteins, spider-web proteins, plant-derived adhesives, adhesives extracted from marine animals, insect- derived adhesives, fibrin, fibroin, Mytilus edulis foot protein (mefpi , "mussel adhesive protein"), other mussel adhesive proteins, proteins and peptides with glycine-rich blocks, proteins and peptides with poly-alanine blocks, and silks.

7. A functionalized AFM tip of claims 5-6, wherein one or more cell attachment factor is chosen from the group comprising ankyrins, cadherins, connexins, dermatan sulphate, entactin, fibrin, fibronectin, glycolipids, glycophorin, glycoproteins, heparan sulphate, heparin sulphate, hyaluronic acid, immunglobulins, keratan sulphate, integrins, laminins, N-CAMs (Calcium independent Adhesive Molecules), proteoglycans, spektrin, vinculin, and vitronectin.

8. A functionalized AFM tip of claims 5-7, wherein one or more biopolymer is chosen from the group comprising alginates, Amelogenins, cellulose, chitosan, collagen, gelatins, oligosaccharides, and pectin.

9. A functionalized AFM tip of claims 5-8, wherein one or more blood protein is chosen from the group comprising albumin, albumen, cytokines, factor IX, factor V, factor VII, factor VIII, factor X, factor Xl, factor XII, factor XIII, hemoglobins (with or without iron), immunglobulins (antibodies), fibrin, platelet derived growth factors (PDGFs), plasminogen, thrombospondin, and transferrin.

10. A functionalized AFM tip of claims 5-9, wherein one or more enzyme is chosen from the group comprising abzymes, adenylate cyclase, alkaline phosphatase, carboxylases, collagenases, cyclooxygenase, hydrolases, isomerases, ligases, lyases, metallo-matrix proteases, nucleases, oxidoreductases, peptidases, peptide hydrolase, peptidyl transferase, phospholipase, proteases, sucrase-isomaltase, TIMPs, and transferases.

11. A functionalized AFM tip of claims 5-10, wherein one or more extracellular matrix protein or biomolecule is chosen from the group comprising ameloblastic amelogenins, collagens (I to XII), dentin-sialo-protein (DSP), dentin- sialo-phospho-protein (DSPP), elastins, enamelin, fibrins, fibronectins, keratins (1 to 20), laminins, tuftelin, carbohydrates, chondroitin sulphate, heparan sulphate, heparin sulphate, hyaluronic acid, lipids and fatty acids, and lipopolysaccarides.

12. A functionalized AFM tip of claims 5-11 , wherein one or more growth factor or hormone is chosen from the group comprising activins; Amphiregulin (AR); Angiopoietins (Ang 1 to 4); Apo3 (a weak apoptosis inducer also known as TWEAK, DR3, WSL-1 , TRAMP or LARD); Betacellulin (BTC); Basic Fibroblast Growth Factor (bFGF, FGF-b); Acidic Fibroblast Growth Factor (aFGF, FGF-a); 4-1 BB Ligand; Brain-derived Neurotrophic Factor (BDNF); Breast and Kidney derived Bolokine (BRAK); Bone Morphogenic Proteins (BMPs); B-Lymphocyte Chemoattractant/B cell Attracting Chemokine 1 (BLC/BCA-1 ); CD27L (CD27 ligand); CD30L (CD30 ligand); CD40L (CD40 ligand); A Proliferation-inducing Ligand (APRIL); Cardiotrophin-1 (CT-1); Ciliary Neurotrophic Factor (CNTF); Connective Tissue Growth Factor (CTGF); Cytokines; 6-cysteine Chemokine (ΘCkine); Epidermal Growth Factors (EGFs); Eotaxin (Eot); Epithelial Cell-derived Neutrophil Activating Protein 78 (ENA-78); Erythropoietin (Epo); Fibroblast Growth Factors (FGF 3 to 19); Fractalkine; Glial-derived Neurotrophic Factors (GDNFs); Glucocorticoid- induced TNF Receptor Ligand (GITRL); Granulocyte Colony Stimulating Factor (G- CSF); Granulocyte Macrophage Colony Stimulating Factor (GM-CSF); Granulocyte Chemotactic Proteins (GCPs); Growth Hormone (GH); I-309; Growth Related Oncogene (GRO); lnhibins (Inh); Interferon-inducible T-cell Alpha Chemoattractant (I-TAC); Fas Ligand (FasL); Heregulins (HRGs); Heparin-Binding Epidermal Growth Factor-Like Growth Factor (HB-EGF); fms-like Tyrosine Kinase 3 Ligand (Flt-3L); Hemofiltrate CC Chemokines (HCC-1 to 4); Hepatocyte Growth Factor (HGF); Insulin; Insulin-like Growth Factors (IGF 1 and 2); Interferon-gamma Inducible Protein 10 (IP-10); lnterleukins (IL 1 to 18); Interferon-gamma (IFN-gamma); Keratinocyte Growth Factor (KGF); Keratinocyte Growth Factor-2 (FGF-10); Leptin (OB); Leukemia Inhibitory Factor (LIF); Lymphotoxin Beta (LT-B); Lymphotactin (LTN); Macrophage-Colony Stimulating Factor (M-CSF); Macrophage-derived Chemokine (MDC); Macrophage Stimulating Protein (MSP); Macrophage Inflammatory Proteins (MIPs); Midkine (MK); Monocyte Chemoattractant Proteins (MCP-1 to 4); Monokine Induced by IFN-gamma (MIG); MSX 1 ; MSX 2; Mullerian Inhibiting Substance (MIS); Myeloid Progenitor Inhibitory Factor 1 (MPIF-1 ); Nerve Growth Factor (NGF); Neurotrophins (NTs); Neutrophil Activating Peptide 2 (NAP- 2); Oncostatin M (OSM); Osteocalcin; OP-1 ; Osteopontin; OX40 Ligand; Platelet derived Growth Factors (PDGF aa, ab and bb); Platelet Factor 4 (PF4); Pleiotrophin (PTN); Pulmonary and Activation-regulated Chemokine (PARC); Regulated on Activation, Normal T-cell Expressed and Secreted (RANTES); Sensory and Motor Neuron-derived Factor (SMDF); Small Inducible Cytokine Subfamily A Member 26 (SCYA26); Stem Cell Factor (SCF); Stromal Cell Derived Factor 1 (SDF-1 ); Thymus and Activation-regulated Chemokine (TARC); Thymus Expressed Chemokine (TECK); TNF and ApoL-related Leukocyte-expressed Ligand-1 (TALL- 1 ); TNF-related Apoptosis Inducing Ligand (TRAIL); TNF-related Activation Induced Cytokine (TRANCE); Lymphotoxin Inducible Expression and Competes with HSV Glycoprotein D for HVEM T-lymphocyte receptor (LIGHT); Placenta Growth Factor (PIGF); Thrombopoietin (Tpo); Transforming Growth Factors (TGF alpha, TGF beta 1 , TGF beta 2); Tumor Necrosis Factors (TNF alpha and beta); Vascular Endothelial Growth Factors (VEGF-A₁B₁C and D).

13. A functionalized AFM tip of claims 5-12, wherein one or more 2'- deoxy nucleic acid is chosen from the group comprising A-DNA; B-DNA; artificial chromosomes carrying mammalian DNA (YACs); chromosomal DNA; circular DNA; cosmids carrying mammalian DNA; DNA; Double-stranded DNA (dsDNA); genomic DNA; hemi-methylated DNA; linear DNA; mammalian cDNA (complimentary DNA; DNA copy of RNA); mammalian DNA; methylated DNA; mitochondrial DNA; phages carrying mammalian DNA; phagemids carrying mammalian DNA; plasmids carrying mammalian DNA; plastids carrying mammalian DNA; recombinant DNA; restriction fragments of mammalian DNA; retroposons carrying mammalian DNA; single- stranded DNA (ssDNA); transposons carrying mammalian DNA; T-DNA; and viruses carrying mammalian DNA; Z-DNA.

14. A functionalized AFM tip of claims 5-13, wherein one or more ribonucleic acid is chosen from the group comprising acetylated transfer RNA (activated tRNA, charged tRNA); circular RNA; linear RNA; mammalian heterogeneous nuclear RNA (hnRNA), mammalian messenger RNA (mRNA); mammalian RNA; mammalian ribosomal RNA (rRNA); mammalian transport RNA (tRNA); mRNA; poly-adenylated RNA; ribosomal RNA (rRNA); recombinant RNA; retroposons carrying mammalian RNA ; ribozymes; transport RNA (tRNA); and viruses carrying mammalian RNA.

15. A functionalized AFM tip of claims 5,-14 wherein one or more receptor is chosen from the group comprising the CD class of receptors CD; EGF receptors; FGF receptors; Fibronectin receptor (VLA-5); Growth Factor receptor, IGF Binding Proteins (IGFBP 1 to 4); lntegrins (including VLA 1-4); Laminin receptor; PDGF receptors; Transforming Growth Factor alpha and beta receptors; BMP receptors; Fas; Vascular Endothelial Growth Factor receptor (Flt-1 ); and Vitronectin receptor.

16. A functionalized AFM tip of claims 5-15, wherein one or more synthetic DNA is chosen from the group comprising A-DNA; antisense DNA; B- DNA; complimentary DNA (cDNA); chemically modified DNA; chemically stabilized DNA; DNA ; DNA analogues ; DNA oligomers; DNA polymers; DNA-RNA hybrids; double-stranded DNA (dsDNA); hemi-methylated DNA; methylated DNA; single- stranded DNA (ssDNA); recombinant DNA; triplex DNA; T-DNA; and Z-DNA.

17. A functionalized AFM tip of claims 5-16, wherein one or more synthetic RNA is chosen from the group comprising antisense RNA; chemically modified RNA; chemically stabilized RNA; heterogeneous nuclear RNA (hnRNA); messenger RNA (mRNA); ribozymes; RNA; RNA analogues; RNA-DNA hybrids; RNA oligomers; RNA polymers; ribosomal RNA (rRNA); and transport RNA (tRNA).

18. A functionalized AFM tip of claims 5-17, wherein one or more synthetic biopolymer is chosen from the group comprising cationic and anionic liposomes; cellulose acetate; hyaluronic acid; polylactic acid; polyglycol alginate; polyglycolic acid; poly-prolines; polysaccharides.

19. A functionalized AFM tip of claims 5-18, wherein one or more synthetic biopolymer is chosen from the group comprising decapeptides containing DOPA and/or diDOPA; peptides with sequence "Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys"; peptides where Pro is substituted with hydroxyproline; peptides where one or more Pro is substituted with DOPA; peptides where one or more Pro is substituted with diDOPA; peptides where one or more Tyr is substituted with DOPA; peptide hormones; peptide sequences based on the above listed extracted proteins; peptides containing an RGD (Arg GIy Asp) motif.

20. A functionalized AFM tip of claims 5-19, wherein one or more synthetic enzyme inhibitor is chosen from the group comprising pepstatin, poly- prolines, D-sugars, D-aminocaids, cyanide, diisopropyl fluorophosphates (DFP), metal ions, N-tosyl-l-phenylalaninechloromethyl ketone (TPCK), Physostigmine, Parathion, and penicillin.

21. A functionalized AFM tip of claims 5-20, wherein one or more synthetic or extracted vitamin is chosen from the group comprising biotin; calciferol (vitamin D's; vital for bone mineralisation); citrin; folic acid; niacin; nicotinamide; nicotinamide adenine dinucleotide (NAD, NAD+); nicotinamide adenine dinucleotide phosphate (NADP, NADPH); NAD+retinoic acid (vitamin A); riboflavin; vitamin B's; vitamin C; vitamin E; and vitamin K's.

22. A functionalized AFM tip of claims 5-21 , wherein one or more pharmaceuticals is chosen from the group comprising antibiotics, cyclooxygenase inhibitors, hormones, inflammation inhibitors, NSAI D's, painkillers, prostaglandin synthesis inhibitors, steroids, and tetracycline.

23. A functionalized AFM tip of claims 5-22, wherein one or more biomolecules is chosen from the group comprising adenosine di-phosphate (ADP); adenosine mono-phosphate (AMP); adenosine tri-phosphate (ATP); amino acids; cyclic AMP (cAMP); 3,4-dihydroxyphenylalanine (DOPA); 5'-di(dihydroxyphenyl-l_- alanine (diDOPA); diDOPA quinone; DOPA-like o-diphenols; fatty acids; glucose; hydroxyproline; nucleosides; nucleotides (RNA and DNA bases); prostaglandin; sugars; sphingosine 1 -phosphate; and rapamycin.

24. A functionalized AFM tip of any of the preceding claims, wherein the biological material comprises a serotonin hybrid film.

25. A functionalized AFM tip of any of the preceding claims, wherein the organic material comprises a film containing peptide bonds.

26. A functionalized AFM tip of any of the preceding claims, wherein the organic material is chosen from a peptide film and a polyamide film.

27. A functionalized AFM tip of any of the preceding claims, wherein the organic-inorganic hybrid material is chosen from a glycine hybrid film, a 4- aminobenzoic acid hybrid film, and a 4-aminobenzophenone hybrid film.

28. A functionalized AFM tip of claims any of the preceding claims, wherein the bone-like material is chosen from calcium phosphate and Ca₂(PO₄)F.

29. A functionalized AFM tip of claims any of the preceding claims, wherein the implant-like material comprises one or more sodium oxides, silicate, calcium oxides, calcium sulfates, calcium phosphates, calcium carbonates, hydroxyapatite, hydrides, vanadium, platinum, hafnium, gold hydroxide, fluorides and oxides of titanium, ferro-titanium alloys, and tantalum metals.

30. A method of coating an AFM tip comprising using ALD to coat said tip with a biological, organic, organic-inorganic hybrid, bone-like, or implant-like material.

31. The method of claim 30, comprising exposing said AFM tip to a precursor comprising a reactive inorganic species in an inert environment, optionally purging said environment with an inert gas, exposing said tip to a biological material in an inert environment, and optionally purging said environment with an inert solvent.

32. The method of claims 30-31 , wherein the biological material is chosen from neurotransmitters, ligands, amino acids, nucleic acids, antibodies, sugars, peptides, proteins, fatty acids, spingolipids, and lipids.

33. The method of claims 30-32, wherein the biological material is chosen from bioadhesives, cell attachment factors, biopolymers, blood proteins, enzymes, extracellular matrix proteins and biomolecules, growth factors and hormones, 2'- deoxy-nucleic acids, ribo-nucleic acids, receptors, synthetic biomolecules, synthetic DNA, synthetic RNA, synthetic biopolymers, synthetic peptides, recombinant proteins, synthetic enzyme inhibitors, synthetic and extracted vitamins, pharmaceuticals, and biologically active ions.

34. The method of claim 33, wherein the bioadhesive is chosen from marine mussel adhesive proteins, fibrin-like proteins, spider-web proteins, plant- derived adhesives, adhesives extracted from marine animals, insect-derived adhesives, fibrin, fibroin, Mytilus edulis foot protein (rnefpi , "mussel adhesive protein"), other mussel adhesive proteins, proteins and peptides with glycine-rich blocks, proteins and peptides with poly-alanine blocks, and silks.

35. The method of claims 33-34, wherein the cell attachment factor is chosen from ankyrins, cadherins, connexins, dermatan sulphate, entactin, fibrin, fibronectin, glycolipids, glycophorin, glycoproteins, heparan sulphate, heparin sulphate, hyaluronic acid, immunglobulins, keratan sulphate, integrins, laminins, N- CAMs (Calcium independent Adhesive Molecules), proteoglycans, spektrin, vinculin, and vitronectin.

36. The method of claims 33-35, wherein the biopolymer is chosen from alginates, Amelogenins, cellulose, chitosan, collagen, gelatins, oligosaccharides, and pectin.

37. The method of claims 33-36, wherein the blood protein is chosen from albumin, albumen, cytokines, factor IX, factor V, factor VII, factor VIII, factor X, factor Xl, factor XII, factor XIII, hemoglobins (with or without iron), immunglobulins (antibodies), fibrin, platelet derived growth factors (PDGFs), plasminogen, thrombospondin, and transferrin.

38. The method of claims 33-37, wherein the enzyme is chosen from abzymes, adenylate cyclase, alkaline phosphatase, carboxylases, collagenases, cyclooxygenase, hydrolases, isomerases, ligases, lyases, metallo-matrix proteases, nucleases, oxidoreductases, peptidases, peptide hydrolase, peptidyl transferase, phospholipase, proteases, sucrase-isomaltase, TIMPs, and transferases.

39. The method of claims 33-38, wherein the extracellular matrix protein or biomolecule is chosen from ameloblastin, amelogenins, collagens (I to XII), dentin-sialo-protein (DSP), dentin-sialo-phospho-protein (DSPP), elastins, enamelin, fibrins, fibronectins, keratins (1 to 20), laminins, tuftelin, carbohydrates, chondroitin sulphate, heparan sulphate, heparin sulphate, hyaluronic acid, lipids and fatty acids, and lipopolysaccarides.

40. The method of claims 33,-39 wherein the growth factor or hormone is chosen from activins; Amphiregulin (AR); Angiopoietins (Ang 1 to 4); Apo3 (a weak apoptosis inducer also known as TWEAK, DR3, WSL-1 , TRAMP or LARD); Betacellulin (BTC); Basic Fibroblast Growth Factor (bFGF, FGF-b); Acidic Fibroblast Growth Factor (aFGF, FGF-a); 4-1 BB Ligand; Brain-derived Neurotrophic Factor (BDNF); Breast and Kidney derived Bolokine (BRAK); Bone Morphogenic Proteins (BMPs); B-Lymphocyte Chemoattractant/B cell Attracting Chemokine 1 (BLC/BCA-1 ); CD27L (CD27 ligand); CD30L (CD30 ligand); CD40L (CD40 ligand); A Proliferation-inducing Ligand (APRIL); Cardiotrophin-1 (CT-1 ); Ciliary Neurotrophic Factor (CNTF); Connective Tissue Growth Factor (CTGF); Cytokines; 6-cysteine Chemokine (6Ckine); Epidermal Growth Factors (EGFs); Eotaxin (Eot); Epithelial Cell-derived Neutrophil Activating Protein 78 (ENA-78); Erythropoietin (Epo); Fibroblast Growth Factors (FGF 3 to 19); Fractalkine; GMaI- derived Neurotrophic Factors (GDNFs); Glucocorticoid-induced TNF Receptor Ligand (GITRL); Granulocyte Colony Stimulating Factor (G-CSF); Granulocyte Macrophage Colony Stimulating Factor (GM-CSF); Granulocyte Chemotactic Proteins (GCPs); Growth Hormone (GH); I-309; Growth Related Oncogene (GRO); lnhibins (Inh); Interferon-inducible T-cell Alpha Chemoattractant (I-TAC); Fas Ligand (FasL); Heregulins (HRGs); Heparin-Binding Epidermal Growth Factor-Like Growth Factor (HB-EGF); fms-like Tyrosine Kinase 3 Ligand (Flt-3L); Hemofiltrate CC Chemokines (HCC-1 to 4); Hepatocyte Growth Factor (HGF); Insulin; Insulin- like Growth Factors (IGF 1 and 2); Interferon-gamma Inducible Protein 10 (IP-10); lnterleukins (IL 1 to 18); Interferon-gamma (IFN-gamma); Keratinocyte Growth Factor (KGF); Keratinocyte Growth Factor-2 (FGF-10); Leptin (OB); Leukemia Inhibitory Factor (LIF); Lymphotoxin Beta (LT-B); Lymphotactin (LTN); Macrophage- Colony Stimulating Factor (M-CSF); Macrophage-derived Chemokine (MDC); Macrophage Stimulating Protein (MSP); Macrophage Inflammatory Proteins (MIPs); Midkine (MK); Monocyte Chemoattractant Proteins (MCP-1 to 4); Monokine Induced by IFN-gamma (MIG); MSX 1 ; MSX 2; Mullerian Inhibiting Substance (MIS); Myeloid Progenitor Inhibitory Factor 1 (MPIF-1 ); Nerve Growth Factor (NGF); Neurotrophins (NTs); Neutrophil Activating Peptide 2 (NAP-2); Oncostatin M (OSM); Osteocalcin; OP-1 ; Osteopontin; OX40 Ligand; Platelet derived Growth Factors (PDGF aa, ab and bb); Platelet Factor 4 (PF4); Pleiotrophin (PTN); Pulmonary and Activation-regulated Chemokine (PARC); Regulated on Activation, Normal T-cell Expressed and Secreted (RANTES); Sensory and Motor Neuron- derived Factor (SMDF); Small Inducible Cytokine Subfamily A Member 26 (SCYA26); Stem Cell Factor (SCF); Stromal Cell Derived Factor 1 (SDF-1 ); Thymus and Activation-regulated Chemokine (TARC); Thymus Expressed Chemokine (TECK); TNF and ApoL-related Leukocyte-expressed Ligand-1 (TALL- 1 ); TNF-related Apoptosis Inducing Ligand (TRAIL); TNF-related Activation Induced Cytokine (TRANCE); Lymphotoxin Inducible Expression and Competes with HSV Glycoprotein D for HVEM T-lymphocyte receptor (LIGHT); Placenta Growth Factor (PIGF); Thrombopoietin (Tpo); Transforming Growth Factors (TGF alpha, TGF beta 1 , TGF beta 2); Tumor Necrosis Factors (TNF alpha and beta); Vascular Endothelial Growth Factors (VEGF-A₁B₁C and D).

41. The method of claims 33-40, wherein the 2'-deoxy nucleic acid is chosen from A-DNA; B-DNA; artificial chromosomes carrying mammalian DNA (YACs); chromosomal DNA; circular DNA; cosmids carrying mammalian DNA; DNA; Double-stranded DNA (dsDNA); genomic DNA; hemi-methylated DNA; linear DNA; mammalian cDNA (complimentary DNA; DNA copy of RNA); mammalian DNA; methylated DNA; mitochondrial DNA; phages carrying mammalian DNA; phagemids carrying mammalian DNA; plasmids carrying mammalian DNA; plastids carrying mammalian DNA; recombinant DNA; restriction fragments of mammalian DNA; retroposons carrying mammalian DNA; single-stranded DNA (ssDNA); transposons carrying mammalian DNA; T-DNA; and viruses carrying mammalian DNA; Z-DNA.

42. The method of claims 33-41 , wherein the ribo-nucleic acid is chosen from acetylated transfer RNA (activated tRNA, charged tRNA); circular RNA; linear RNA; mammalian heterogeneous nuclear RNA (hnRNA), mammalian messenger RNA (mRNA); mammalian RNA; mammalian ribosomal RNA (rRNA); mammalian transport RNA (tRNA); mRNA; poly-adenylated RNA; ribosomal RNA (rRNA); recombinant RNA; retroposons carrying mammalian RNA ; ribozymes; transport RNA (tRNA); and viruses carrying mammalian RNA.

43. The method of claims 33,-42 wherein the receptor is chosen from the group comprising the CD class of receptors CD; EGF receptors; FGF receptors; Fibronectin receptor (VLA-5); Growth Factor receptor, IGF Binding Proteins (IGFBP 1 to 4); lnteghns (including VLA 1-4); Laminin receptor; PDGF receptors; Transforming Growth Factor alpha and beta receptors; BMP receptors; Fas; Vascular Endothelial Growth Factor receptor (Flt-1 ); and Vitronectin receptor.

44. The method of claims 33-43, wherein the synthetic DNA is chosen from A-DNA; antisense DNA; B-DNA; complimentary DNA (cDNA); chemically modified DNA; chemically stabilized DNA; DNA ; DNA analogues ; DNA oligomers; DNA polymers; DNA-RNA hybrids; double-stranded DNA (dsDNA); hemi- methylated DNA; methylated DNA; single-stranded DNA (ssDNA); recombinant DNA; triplex DNA; T-DNA; and Z-DNA.

45. The method of claims 33,-44 wherein the synthetic RNA is chosen from antisense RNA; chemically modified RNA; chemically stabilized RNA; heterogeneous nuclear RNA (hnRNA); messenger RNA (mRNA); ribozymes; RNA; RNA analogues; RNA-DNA hybrids; RNA oligomers; RNA polymers; ribosomal RNA (rRNA); and transport RNA (tRNA).

46. The method of claims 33-45, wherein the synthetic biopolymer is chosen from cationic and anionic liposomes; cellulose acetate; hyaluronic acid; polylactic acid; polyglycol alginate; polyglycolic acid; poly-prolines; polysaccharides.

47. The method of claims 33-46, wherein the synthetic enzyme inhibitor is chosen from pepstatin, poly-prolines, D-sugars, D-aminocaids, cyanide, diisopropyl fluorophosphates (DFP), metal ions, N-tosyl-l-phenylalaninechloromethyl ketone (TPCK), Physostigmine, Parathion, and penicillin.

48. The method of claims 33-47, wherein the synthetic or extracted vitamin is chosen from biotin; calciferol (vitamin D's; vital for bone mineralisation); citrin; folic acid; niacin; nicotinamide; nicotinamide adenine dinucleotide (NAD, NAD+); nicotinamide adenine dinucleotide phosphate (NADP, NADPH); NAD+retinoic acid (vitamin A); riboflavin; vitamin B's; vitamin C; vitamin E; and vitamin K¹S.

49. The method of claims 33-48, wherein the pharmaceutical is chosen from antibiotics, cyclooxygenase inhibitors, hormones, inflammation inhibitors, NSAID's, painkillers, prostaglandin synthesis inhibitors, steroids, and tetracycline.

50. The method of claims 33-49, wherein the biologically active ions is chosen from adenosine di-phosphate (ADP); adenosine mono-phosphate (AMP); adenosine tri-phosphate (ATP); amino acids; cyclic AMP (cAMP); 3,4- dihydroxyphenylalanine (DOPA); 5'-di(dihydroxyphenyl-L-alanine (diDOPA); diDOPA quinone; DOPA-like o-diphenols; fatty acids; glucose; hydroxyproline; nucleosides; nucleotides (RNA and DNA bases); prostaglandin; sugars; sphingosine 1 -phosphate; and rapamycin.

51. The method of claims 30-50, wherein the biological material is serotonin.

52. The method of claims 31-52, wherein the precursor is chosen from BCI₃, BBr₃, B(OMe)₃, AICI₃, AIBr₃, AIMe₂CI, AIMe₂C₁Pr, AIMe₂H, AI(OEt)₃, AI(CPr)₃, a trialkyl aluminum, GaCI₃, GaMe₃, GaCI, GaBr, GaI, GaMe₃, GaEt₃, Ga(acac)₃, Ga, GaEt₂CI, GaEt₂Me, InCI₃, InMe₃, InEt₃, ln(acac)₃, In,, InEtMe₂, InCI, InCIMe₂, CF₃, SiCI₄, SiCI₃H, SiCI₂H₂, SiH₄, Si₂H₆, SiCI₃H, Si(OEt)₄, Si(O"Bu)₄, ('BuO)₃SiOH, GeCI₄, GeMe₂H₂, GeEt₂H₂, GeH₄, Ge₂H₆, SnCI₄, SnEt₄, SnMe₄, SnI₄, SnCI₄, PbBr₂, PbI₂, Pb(OAc)₂, Pb(O⁴Bu)₂, Pb(thd)₂, Pb(detdc)₂, YCp₃, Y(CpMe)₃, Y(thd)₃, Cd, CdMe₂, CdCI₂, PCI₃, POCI₃, SbCI₅, Bi(Ph)₃, Bi[N(SiMe₃)₂]₃ TiCI₄, TiI₄, Ti(NMe₂J₄, Ti(NEt₂J₄, Ti(NMeEt)₄, Ti(CPr)₄, Ti(OEt)₄, ZrCI₄, ZrI₄, ZrCp₂CI₂, ZrCp₂Me₂, Zr(O'Pr)₂(dmae)₂, Zr(O^fBu)₄, Zr(dmae)₄, Zr(thd)₄, Zr(NMe₂J₄, Zr(NEtMe)₄, Zr(NEt₂J₄, HfCI₄, HfCI₂[N(SiMe₃)₂]₂, HfI₄, Hf(O^fBu)₄, Hf(θ'Bu)₂(mmp)₂, Hf(mmp)₄, Hf(ONEt₂J₄, Hf(NEt₂)* Hf(NEtMe)₄, Hf[N(SiMe₃)₂]₂CI₂, Hf(NOa)₄, VOCI₃, VO(O',Pr)₃, VO(acac)₂, Nb(OEt)₅, NbCI₅, TaF₅, TaCI₅, TaBr₅, TaI₅, Ta(OEt)₅, Ta(NMe₂)₅, Ta(NEt₂J₅, Ta(NEt)(NEt₂J₃, Ta(N'Bu)(NEt₂)₃, Ta(N^Bu)(NEtMe)₃, CrO₂CI₂, Cr(acac)₃, MoCI₅, WF₆, WOCI₄, WF_xOy, W(N^fBu)₂(NMe₂)₂, Mn(thd)₃, MnCI₂, Mn, Fe(acac)₃, FeCI₃, Fe(thd)₃, Fe(^BuAMD)₂, Ru(CpEt)₂, RuCp₂, Ru(Od)₃, Ru(thd)₃, Co('PrAMD)₂, Co(acac)₃, Co(thd)₂, lr(acac)Ca(thd)₃, NiCp₂, Ni(acac)₂, Ni(apo)₂, Ni(dmg)₂, Ni('PrAMD)_2, Ni(thd)₂, Pd(thd)₂, Pd(hfac)₂, Pt(CpMe)Me₃, Pt(acac)₂, Cu(acac)₂, Cu(thd)₂, Cu(hfac)₂, CuCI, Cu('PrAMD), ZnCI₂, ZnMe₂, ZnEt₂, Zn(OAc)₂, Zn, Zn[N(SiMe₃)J, HgMe₂, Mg, Mg(Cp)₂, Mg(thd)₂, Ca(thd)₂, CaCp₂, Sr(Cp¹Pr₃J₂, Sr(thd)₂, Sr(methd)₂₎ Sr(CpMe₅J₂, Ba(CpMe₅J₂, Ba(thd)₂, ScCp₃, Sc(thd)₃, La[N(SiMe₃J₂J₃, La(PrAMD)₃, La(thd)₃, Ce(thd)₄, Ce(thd)₃phen, Pr[N(SiMe₃)₂]₃, Nd(thd)₃, Sm(thd)₃, Eu(thd)₃, Gd(thd)₃, Dy(thd)₃, Ho(thd)₃, Er(thd)₃, Tm(thd)₃₎ and Lu[Co(SiMe₃)]₂CI.

53. The method of claims 30-52, comprising exposing said AFM tip to a precursor gas comprising molecules having two anhydride moieties or having two acyl chloride moieties, optionally purging with an inert gas, exposing said tip to a precursor gas comprising molecules having two amino groups, and optionally purging with an inert solvent.

54. The method of claims 30-52, comprising exposing said AFM tip to a first precursor gas comprising molecules having two amino groups, optionally purging with an inert gas, exposing said tip to precursor gas comprising molecules two anhydride moieties and/or molecules having two acyl chloride moieties, and optionally purging with an inert solvent.

55. The method of claims 54 and 55 wherein the precursor is chosen from 1 ,2,4,5-benzenetetracarboxylic anhydride and nonanedioyl chloride.

56. The method of claims 54, 55, and 56 wherein said precursor is chosen from ethylenediamine, 1 ,6,-diaminohexame, 1 ,4-phenylenediamine, and 4,4'-oxydianiline.

57. The method of claims 30-52, comprising exposing said AFM tip to a precursor gas comprising molecules of the type H₂N-R-C≡C-R' in an inert environment, optionally purging said environment with an inert gas, exposing said tip to a precursor gas comprising ozone in an inert environment, and optionally purging said environment with an inert gas.

58. The method of claims 30-52, comprising exposing said AFM tip to a precursor gas comprising molecules of the type H₂N-R-C=C-R' in an inert environment, optionally purging said environment with an inert gas, exposing said tip to a precursor gas comprising ozone in an inert environment, and optionally purging said environment with an inert gas.

59. The method of claim 58 wherein the AFM tip surface is first terminated with carboxylic anhydride moieties.

60. The method of claims 30-52, comprising exposing said AFM tip to an inorganic precursor in an inert environment, optionally purging said environment with an inert gas, exposing said tip to an organic precursor in an inert environment, and optionally purging said environment with an inert gas.

61. The method of claim 60, wherein the organic precursor is chosen from glycine, 4-aminobenzoic acid, and 4-aminobenzophenone.

62. The method of claim 60 and 61 , wherein the inorganic precursor is chosen from BCI₃, BBr₃, B(OMe)₃, AICI₃, AIBr₃, AIMe₂CI, AIMe₂O',Pr, AIMe₂H, AI(OEt)₃, AI(CPr)₃, a trialkyl aluminum, GaCI₃, GaMe₃, GaCI, GaBr, GaI, GaMe₃, GaEt₃, Ga(acac)₃, Ga, GaEt₂CI, GaEt₂Me, InCI₃, InMe₃, InEt₃, ln(acac)₃, In₁, InEtMe₂, InCI, InCIMe₂, CF₃, SiCI₄, SiCI₃H, SiCI₂H₂, SiH₄, Si₂H₆, SiCI₃H, Si(OEt)₄, Si(O⁰Bu)₄, ('BuO)₃SiOH, GeCI₄, GeMe₂H₂, GeEt₂H₂, GeH₄, Ge₂H₆, SnCI₄, SnEt₄, SnMe₄, SnI₄, SnCI₄, PbBr₂, PbI₂, Pb(OAc)₂, Pb(O¹Bu)₂, Pb(thd)₂, Pb(detdc)₂, YCp₃, Y(CpMe)₃, Y(thd)₃, Cd, CdMe₂, CdCI₂, PCI₃, POCI₃, SbCI₅, Bi(Ph)₃, Bi[N(SiMe₃)₂]₃ TiCI₄, TiI₄, Ti(NMe₂J₄, Ti(NEt₂J₄, Ti(NMeEt)₄, Ti(CPr)₄, Ti(OEt)₄, ZrCI₄, ZrI₄, ZrCp₂CI₂, ZrCp₂Me₂, Zr(O'Pr)₂(dmae)₂, Zr(O^fBu)₄, Zr(dmae)₄, Zr(thd)₄, Zr(NMe₂)₄, Zr(NEtMe)₄, Zr(NEt₂J₄, HfCI₄, HfCI₂[N(SiMe₃)₂]₂, HfI₄, Hf(O^fBu)₄, Hf(O^fBu)₂(mmp)₂, Hf(mmp)₄, Hf(ONEt₂J₄, Hf(NEt₂J₄, Hf(NEtMe)₄, Hf[N(SiMe₃)₂]₂CI₂, Hf(NO₃J₄, VOCI₃, VO(O',Pr)₃, VO(acac)₂, Nb(OEt)₅, NbCI₅, TaF₅, TaCI₅, TaBr₅, TaI₅, Ta(OEt)₅, Ta(NMe₂)_S, Ta(NEt₂J₅, Ta(NEt)(NEt₂J₃, Ta(N'Bu)(NEt₂)₃, Ta(N^Bu)(NEtMe)₃, CrO₂CI₂, Cr(acac)₃, MoCI₅, WF₆, WOCI₄, WF_xO_y, W(N'Bu)₂(NMe₂)₂, Mn(thd)₃, MnCI₂, Mn, Fe(acac)₃, FeCI₃, Fe(thd)₃, Fe(^fBuAMD)_2, Ru(CpEt)₂, RuCp₂, Ru(Od)₃, Ru(thd)₃, Co('PrAMD)_2, Co(acac)₃, Co(thd)₂₎ lr(acac)Ca(thd)₃, NiCp₂, Ni(acac)₂, Ni(apo)₂, Ni(dmg)₂, Ni(^rAMD)₂, Ni(thd)₂, Pd(thd)₂, Pd(hfac)₂, Pt(CpMe)Me₃, Pt(acac)₂, Cu(acac)₂, Cu(thd)₂, Cu(hfac)₂, CuCI, Cu('PrAMD), ZnCI₂, ZnMe₂, ZnEt₂, Zn(OAc)₂, Zn, Zn[N(SiMe₃)₂], HgMe₂, Mg, Mg(Cp)₂, Mg(thd)₂, Ca(thd)₂, CaCp₂, Sr(Cp'Pr₃)₂, Sr(thd)₂, Sr(methd)₂₎ Sr(CpMe₅J₂, Ba(CpMe₅J₂, Ba(thd)₂, ScCp₃, Sc(thd)₃, La[N(SiMe₃Js]₃, La(¹PrAMD)₃, La(thd)₃, Ce(thd)₄, Ce(thd)₃phen, Pr[N(SiMe₃)₂]₃, Nd(thd)₃, Sm(thd)₃, Eu(thd)₃, Gd(thd)₃, Dy(thd)₃, Ho(thd)₃, Er(thd)₃, Tm(thd)₃, and Lu[Co(SiMe₃)J₂CI.

63. The method of claims 30-52, comprising exposing said AFM tip to a calcium precursor, optionally purging said environment with an inert gas, exposing said tip to a phosphorous precursor in an inert environment, optionally purging said environment with an inert solvent, and exposing said environment to a pulse of either water or NH₄F.

64. The method of claim 63, wherein the calcium precursor is chosen from Ca(Cp)₂ and Ca(thd)₂.

65. The method of claims 64 and 65, wherein the phosphorous precursor is chosen from POCI₃, P(Ph)₃, and (CH₃O)₃PO.

66. The method of claims 30-52, wherein the implant-like material deposited comprises one or more sodium oxides, silicate, calcium oxides, calcium sulfates, calcium phosphates, calcium carbonates, hydroxyapatite, hydrides, vanadium, platinum, hafnium, gold hydroxide, fluorides and oxides of titanium, ferro-titanium alloys, or tantalum metals.

67. The method of claims 30-52 wherein the AFM tip is coated using liquid-phase ALD in an inert atmosphere, comprising exposing said AFM tip to a first precursor containing a reactive inorganic species in an inert solvent, optionally rinsing said tip with an inert solvent, exposing said tip to a biological material dissolved or suspended in an inert solvent, and optionally rinsing said tip with an inert solvent.

68. The method of claims 66 and 67, wherein the inorganic precursor is chosen from BCI₃, BBr₃, B(OMe)₃, AICI₃, AIBr₃, AIMe₂CI, AIMe₂C₁Pr, AIMe₂H, AI(OEt)₃, AI(O"Pr)₃, a trialkyl aluminum, GaCI₃, GaMe₃, GaCI, GaBr, GaI, GaMe₃, GaEt₃, Ga(acac)₃, Ga, GaEt₂CI, GaEt₂Me, InCI₃, InMe₃, InEt₃, ln(acac)₃, In,, InEtMe₂, InCI, InCIMe₂, CF₃, SiCI₄, SiCI₃H, SiCI₂H₂, SiH₄, Si₂H₆, SiCI₃H, Si(OEt)₄, Si(O⁰Bu)₄, ('BuO)₃SiOH, GeCI₄, GeMe₂H₂, GeEt₂H₂, GeH₄, Ge₂H₆, SnCl₄, SnEt₄, SnMe₄, SnI₄, SnCI₄, PbBr₂, PbI₂, Pb(OAc)₂, Pb(O¹Bu)₂, Pb(thd)₂, Pb(detdc)₂, YCp₃, Y(CpMe)₃, Y(thd)₃, Cd, CdMe₂, CdCI₂, PCI₃, POCI₃, SbCI₅, Bi(Ph)₃, Bi[N(SiMe₃)₂]₃ TiCI₄, TiI₄, Ti(NMe₂)₄, Ti(NEt₂)₄, Ti(NMeEt)₄, Ti(CPr)₄, Ti(OEt)₄, ZrCI₄, ZrI₄, ZrCp₂CI₂, ZrCp₂Me₂, Zr(O'Pr)₂(dmae)₂, Zr(O^Bu)₄, Zr(dmae)₄₎ Zr(thd)₄, Zr(NMe₂)₄, Zr(NEtMe)₄, Zr(NEt₂J₄, HfCI₄, HfCI₂[N(SiMe₃)₂]₂, HfI₄, Hf(O^fBu)₄, Hf(O'Bu)₂(mmp)₂, Hf(mmp)_4> Hf(ONEt₂)₄, Hf(NEt₂)₄, Hf(NEtMe)₄, Hf[N(SiMe₃)₂]₂CI₂, Hf(NO₃J₄, VOCI₃, VO(O',Pr)₃, VO(acac)₂, Nb(OEt)₅, NbCI₅, TaF₅, TaCI₅, TaBr₅, TaI₅, Ta(OEt)₅, Ta(NMe₂)_S, Ta(NEt₂)₅, Ta(NEt)(NEt₂)₃, Ta(N^fBu)(NEt₂)₃, Ta(N^fBu)(NEtMe)₃, CrO₂CI₂, Cr(acac)₃, MoCI₅, WF₆, WOCI₄, WF_xOy, W(N^fBu)₂(NMe₂)₂, Mn(thd)₃, MnCI₂, Mn, Fe(acac)₃, FeCI₃, Fe(thd)₃, Fe(^BuAMD)₂, Ru(CpEt)₂, RuCp₂, Ru(Od)₃, Ru(thd)₃, Co('PrAMD)₂, Co(acac)₃, Co(thd)₂, lr(acac)Ca(thd)_3> NiCp₂, Ni(acac)₂, Ni(apo)₂, Ni(dmg)₂, Ni(¹PrAMD)₂, Ni(thd)₂, Pd(thd)₂, Pd(hfac)₂, Pt(CpMe)Me₃, Pt(acac)₂, Cu(acac)₂, Cu(thd)₂, Cu(hfac)₂₎ CuCI, Cu('PrAMD), ZnCI₂, ZnMe₂, ZnEt₂, Zn(OAc)₂, Zn, Zn[N(SiMe₃J₂], HgMe₂, Mg, Mg(Cp)₂, Mg(thd)₂, Ca(thd)₂, CaCp₂, Sr(Cp'Pr₃)₂, Sr(thd)₂, Sr(methd)₂, Sr(CpMe₅J₂, Ba(CpMe₅J₂, Ba(thd)₂, ScCp₃, Sc(thd)₃, La[N(SiMe₃)₂]₃, La('PrAMD)₃, La(thd)₃, Ce(thd)₄, Ce(thd)₃phen, Pr[N(SiMe₃)₂]₃, Nd(thd)₃₎ Sm(thd)₃, Eu(thd)₃, Gd(thd)₃, Dy(thd)₃, Ho(thd)₃, Er(thd)₃, Tm(thd)₃, and Lu[Co(SiMe₃)]₂CI.

69. The method of claims 67 and 68, wherein the inert solvents are each chosen from toluene, hexane, and heptane.

70. The method of claims 30 - 69, wherein said method is repeated 1 - 2000 times.

71. A functionalized AFM tip prepared using the method of claims 30 - 70.

72. A method of studying a surface comprising using an AFM tip as claimed in claims 1 - 29 in an AFM to image and/or probe a sample surface.

73. The method of claim 72, wherein the sample surface comprises a biological, organic, inorganic-hybrid, bone, or implant-like material.

74. The method of claim 72, wherein the sample surface contains a cell, protein, enzyme, antibody, or peptide.

75. The method of claim 72, wherein the sample surface is a bone cell.

76. A method of studying a surface comprising using an AFM tip as claimed in claim 1 - 29 in an AFM to study the intermolecular forces between said tip and a sample surface.

77. The method of claim 76, wherein the sample surface comprises a biological, organic, inorganic-hybrid, bone, or implant-like material.

78. The method of claim 76, wherein the sample surface contains a cell, protein, enzyme, antibody, or peptide.

79. The method of claim 76, wherein the sample surface is a bone cell.

80. A functionalized SPM tip comprising a SPM tip coated with at least one material chosen from biological, organic, inorganic, organic-inorganic hybrid, magnetic/conductive and hard coatings.

81. A functionalized SPM tip of claim 80, wherein the SPM tip is a MFM tip coated in a magnetic/conductive film.

82. The MFM tip of claim 81 , wherein the film is chosen from ferromagnetic, ferrimagnetic and paramagnetic films.

83. The MFM tip of claim 81-82, wherein the magnetic film is (Fe₁Co)₃O₄.

84. The MFM tip of claim 81-83, wherein the (Fe₁Co)₃O₄ film is 5-200 nm thick.

85. A functional ized SPM tip of claim 81 , wherein the SPM tip comprises a hard coating chosen from Tiθ₂, AI₂O₃, ZrO₂, Ti-Nitride, multi-layer and hybrid hard coatings.

86. A SPM probe coated with TiO₂.