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Application note · Scanning probe microscopy

Plasma cleaning and sharpening of AFM tips

The tip decides what an atomic force microscope can resolve. Low-pressure plasma removes organic contamination from AFM probes, makes silicon and silicon-nitride tips hydrophilic, activates them for functionalization — and, in high-speed AFM labs worldwide, sharpens electron-beam-deposited carbon tips to a 1–2 nm apex in about a minute.

  • Tip apex ≈1–2 nm after Ar plasma
  • 20 W, 1–1.5 min, argon
  • Used in 18 published AFM studies
Schematic: an electron-beam-deposited carbon spike on an AFM tip is sharpened by argon plasma in a Tergeo plasma cleaner to an apex radius of about 1 to 2 nanometers
How high-speed AFM labs make ultra-sharp tips: grow a carbon spike in an SEM, then sharpen it with argon plasma.
Why plasma

Three things plasma does for AFM probes

Sharpen carbon tips

Argon plasma etches the amorphous carbon spike grown on a probe by electron-beam deposition (EBD), thinning its end to an apex radius of about 1–2 nm. This is the standard way high-speed AFM (HS-AFM) labs make tips sharp enough to resolve single proteins.

Remove contamination

New cantilevers often arrive coated with organic contamination, such as silicone from the packaging gel. That film changes adhesion and friction forces and can transfer to the sample. A short oxygen or air plasma removes it without solvents or touching the tip.

Hydrophilic, reactive surfaces

Plasma leaves silicon and silicon-nitride surfaces covered in hydroxyl groups. The tip wets easily in liquid imaging, and silane (e.g. APTES) or other coupling chemistry attaches reliably when making functionalized probes.

Published method

Sharpening EBD carbon tips for high-speed AFM

HS-AFM films proteins at video rate with small cantilevers whose built-in tip is too blunt for molecular resolution. Labs therefore grow an amorphous carbon spike on the tip in an SEM (typically from ferrocene or residual hydrocarbons) and sharpen it in a Tergeo plasma cleaner with argon plasma.

Published conditionsValue
GasArgon
ModeDirect (immersion) plasma
RF power15–20 W
Gas flow / pressure10 sccm, or ≈180 mTorr
Time30 s – 1.5 min
ResultApex radius ≈1 nm (several studies); 4 nm → 1.5–2 nm (Ye et al., 2024)

The most-cited recipe: direct mode, Ar 10 sccm, 20 W, 1.5 min.

Why it works

In direct mode the probe sits in the argon glow, and ions remove the amorphous carbon atom by atom. The thin end of the spike loses material from all sides, so the apex gets sharper while the spike stays strong enough to scan. Argon is chemically inert, so the silicon-nitride cantilever and its reflective coating are not oxidized.

Because etching is fast at this scale, power and time matter. The Tergeo's 1 W power steps, recipe memory and real-time plasma intensity reading let a lab repeat the same sharpening on every cantilever — and a gold-coated tip can still be functionalized afterwards, as shown for molecular-recognition HS-AFM (Puppulin et al., 2021).

Cleaning & activation

Cleaning AFM probes and making them hydrophilic

Organic and inorganic contamination on as-received AFM cantilevers is well documented (Lo et al., 1999). It shows up as unstable adhesion in force spectroscopy, drifting friction in lateral force microscopy, and residues transferred to clean samples.

A gentle oxygen, air or water-vapor plasma removes this layer and leaves a hydrophilic, hydroxyl-rich surface on silicon and silicon-nitride probes. The same step activates cantilever surfaces before covalent functionalization — for example, microcantilever sensor arrays were O₂-plasma activated in a Tergeo-Plus before attaching their sensing chemistry (Cooper et al., Biosens. Bioelectron. 2022 — MEMS sensor cantilevers rather than AFM imaging tips).

Plasma-cleaned surfaces slowly pick up airborne hydrocarbons again, so clean or activate probes shortly before use.

Probe typeSuggested starting point
Bare Si or Si₃N₄ — cleaning, hydrophilicO₂, air or H₂O vapor; low power, downstream or pulsed; seconds to a minute
Before silane / APTES functionalizationO₂ or H₂O vapor plasma, then functionalize promptly
EBD carbon tip sharpeningAr, direct mode, 20 W, 10 sccm, 30–90 s
Carbon, diamond-like or polymer-coated tips — cleaning onlyAvoid oxygen; use argon or downstream mode at very low power
Metal-coated (Au, Pt) and magnetic tipsShort, gentle plasma; test first — oxygen can oxidize some coatings

Starting points only. Oxygen plasma also etches carbon, so never use it on a carbon tip you want to keep sharp.

Publications

Published studies that sharpened AFM tips in a Tergeo plasma cleaner

Each study below states in its methods that the electron-beam-deposited carbon tip was sharpened with argon plasma in a Tergeo plasma system (sometimes listed as "PIC Scientific"). The plasma step is part of the methods — most papers are about the proteins imaged. Most come from high-speed AFM groups at Kanazawa University (WPI-NanoLSI) and their collaborators.

StudyJournalTip processing reported
Fukuda & Ando — Faster high-speed atomic force microscopy for imaging of biomolecular processesRev. Sci. Instrum., 2021Ar, 15 W, 1 min
Niina et al. — Rigid-body fitting to AFM images for inferring probe shape and biomolecular structurePLOS Comput. Biol., 2021Ar, direct, 10 sccm, 20 W, 1.5 min
Kodera et al. — Native cyclase-associated protein and actin form a unique 4:4 complexJ. Biol. Chem., 2021Ar, direct, 10 sccm, 20 W, 1.5 min
Puppulin et al. — Macrocyclic peptide-conjugated tip for molecular recognition imaging by HS-AFMACS Appl. Mater. Interfaces, 2021Plasma etched to ≈1 nm apex, then functionalized
Yoshimi et al. — Dynamic mechanisms of CRISPR interference by E. coli CRISPR-Cas3Nat. Commun., 2022Ar, direct, 10 sccm, 20 W, 1.5 min
Kodera & Ando — Guide to studying intrinsically disordered proteins by HS-AFMMethods, 2022Ar RF plasma sharpening of ≈25 nm EBD tips
Hayakawa et al. — Actin-binding domain of Rng2 inhibits actin movement on myosin IILife Sci. Alliance, 2022Ar, ≈180 mTorr, 20 W, 30 s
Kubo et al. — Removing the parachuting artifact using two-way scanning data in HS-AFMBiophys. Physicobiol., 2023Ar, direct, 10 sccm, 20 W, 1.5 min
Tsujioka et al. — Imaging single CaMKII holoenzymes at work by HS-AFMScience Advances, 2023Plasma etched to ≈1 nm apex
Ye et al. — Structural heterogeneity of the ion and lipid channel TMEM16FNat. Commun., 2024Ar plasma; apex 4 nm → 1.5–2 nm
Ngo et al. — Actin structure-dependent cooperative binding of cofilineLife, 2024Ar, ≈180 mTorr, 20 W, 90 s
Sumino et al. — HS-AFM reveals dimer splitting of the GluA2 receptor N-terminal domainACS Nano, 2024Plasma etched to ≈1 nm apex
Takeda et al. — Oligomer-based functions of mitochondrial porinNat. Commun., 2025Ar, direct, 10 sccm, 20 W, 1.5 min
Matsushima et al. — Structural dynamics of mixed-subunit CaMKIIα/β heterododecamersNat. Commun., 2025Plasma etched to ≈1 nm apex
Yamaguchi et al. — Heterohexameric assembly of the LGI1–ADAM22 complexeLife, 2025Plasma etched to ≈1 nm apex
Suzuki et al. — CaMKIIα holoenzymes self-organize into chain-like mesoscale clustersScience Advances, 2026Plasma etched to ≈1 nm apex
Mahmud et al. — Visualization of G3BP1–RNA condensate assembly by HS-AFMInt. J. Mol. Sci., 2026Ar, direct, 10 sccm, 20 W, 1.5 min
Yurtsever et al. — Peptides display multilayered hydration shell substructuresNat. Commun., 2026Ar plasma sharpening of EBD carbon tips (3D-AFM)

Background reference: Lo, Y.-S. et al., Organic and inorganic contamination on commercial AFM cantilevers, Langmuir 15, 6522 (1999).

FAQ

AFM tip plasma cleaning: common questions

Can I plasma clean an AFM tip?

Yes. A short, low-power oxygen, air or water-vapor plasma removes organic contamination from silicon and silicon-nitride tips and makes them hydrophilic. Use argon or a very gentle downstream setting instead for carbon, diamond-like or polymer-coated tips, which oxygen would etch.

What recipe do labs use to sharpen EBD carbon tips?

The most common published recipe is argon plasma in direct mode, 10 sccm, 20 W for 1.5 minutes in a Tergeo plasma cleaner. Other groups report 15 W for 1 minute or about 180 mTorr at 20 W for 30–90 seconds. Apex radii of about 1–2 nm are reported.

Will plasma damage the cantilever or its reflective coating?

Argon does not oxidize the silicon-nitride cantilever or a gold back-side coating, and the published sharpening steps last only seconds to about a minute. Test oxygen-containing recipes on a spare probe first if your tip has a metal or magnetic coating.

How long does a plasma-cleaned tip stay clean?

Airborne hydrocarbons start to re-adsorb within hours. Clean or activate probes shortly before imaging or functionalization, and store them in a clean, closed box.

Which plasma system do I need?

A tabletop Tergeo handles cantilevers and probe boxes and is the system used in the studies above. The Tergeo-EM adds TEM holder ports if your lab also does electron microscopy.

Related: Tergeo plasma cleaner · Argon plasma cleaning · TEM, SEM & FIB sample cleaning · Fragile TEM carbon grids

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