Peak Force Infrared (PFIR) Microscopy

Peak Force Infrared Microscopy enabling simultaneous chemical and mechanical imaging at ~6nm resolution.

What is Peak Force Infrared (PFIR) Microscopy?

PFIR microscopy is action-based microscopy with mechanical detection of photothermal expansion by atomic force microscope (AFM) operated in the peak force tapping mode. It falls under the umbrella of the family of photothermal AFM-IR techniques. Our group at Lehigh University invented PFIR microscopy in 2016-2017.

The underlying mechanism for PFIR (and AFM-IR in general) is to leverage the high force sensitivity of an AFM cantilever to measure the mechanical effect of IR-specific photothermal expansion of the sample. The contact area of the AFM tip can be smaller than the tip radius in PFIR, thus the measurement is achieved from an area much smaller than the optical diffraction limit--a route for super-resolution microscopy.

The unique trait of PFIR among AFM-IR techniques is the AFM tip is predictably in contact and detached from the sample surface under feedback loop control. The sample surface integrity is well preserved. As the cantilever resonances behave differently when in contact or detached from the sample, no residual photothermal signals are carried over from previous tip-sample interactions, thus ensuring no signal spillover between adjacent image pixels and achieving high spatial resolution as high as 6 nm.

The PFIR microscopy has another intrinsic advantage inherited from the peak force tapping mode. Under weak IR illumination, the mechanical properties of modulus and adhesion are simultaneously collected with the IR images, without separate scans. As the peak force tapping mode works well in the fluid, the PFIR microscopy also works in the fluid phase, including water.

The details of the PFIR microscopy are described in our recent Chemical Society Review article.

Operational diagram of single pulse PFIR microscopy
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The operational diagram of the original single pulse PFIR microscopy. The emissions of the laser pulses are synchronized to the tip-sample contact during the peak force tapping cycle.

PFIR Achieves Label-free Chemical Imaging at 6 nm Spatial Resolution

The photothermal responses of the sample are typically extracted from the cantilever response using a data acquisition card, processed in real-time, and used as the PFIR signal. This approach achieves label-free chemical imaging at 6 nm spatial resolution, far surpassing the diffraction limit of conventional optical methods.

PFIR 6nm spatial resolution demonstration
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PFIR achieves 6 nm spatial resolution for label-free chemical imaging

Improvements Since Invention

Since its invention, PFIR microscopy has been improved in several aspects:

  • Multi-pulse PFIR for improvement of signal
  • Total internal reflection and Liquid-phase PFIR microscopy
  • Dual-color PFIR for multiple component imaging
  • PFIR-KPFM for multimodal chemical, mechanical, and electrical imaging

If you are interested in measurement of your samples with PFIR, or building a PFIR microscope in your lab, please contact us. We are open to collaborations.

PFIR improvements overview
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Multi-pulse PFIR application
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Multi-pulse PFIR
Liquid-phase PFIR application
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Liquid-phase PFIR
Dual-color PFIR application
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Dual-color PFIR
PFIR-KPFM application
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PFIR-KPFM

Multi-pulse PFIR Microscopy

One main limitation of the regular PFIR microscopy is its relatively weak signal. The peak force tapping cycle of an AFM is usually a few kHz (1kHz up to 8 kHz), which means that the tip and sample are in contact a few thousand times per second. At each tip-sample contact, a laser pulse is emitted to excite the photothermal expansion of the sample, and the cantilever responses are extracted to be used as a signal.

However, for samples with low photothermal responses or IR absorption, the PFIR signals are often weak.

The implementation of the multi-pulse PFIR is to illuminate multiple laser pulses per peak force tapping cycle. So more photothermal responses are generated per unit time. The resulting method improves the signal quality of PFIR microscopy.

The details of multi-pulse PFIR microscopy are described in our paper Advanced Optical Materials 1901084 (2019).

Multi-pulse PFIR scheme and comparison
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Left: An illustration of the multi-pulse PFIR scheme. Right: comparison of the single-pulse PFIR signal versus multi-pulse PFIR signal on spatial imaging of the phonon polaritons of h-BN.

Liquid-Phase PFIR and Total Internal Reflection PFIR

One limitation of traditional infrared microscopy, aside from the restriction of spatial resolution from Abbe's diffraction limit, is its application in the aqueous phase. Water strongly absorbs mid-IR radiation, making the optical delivery difficult. Water absorption also strongly attenuates optical detection after light-matter interactions, reducing IR signals.

Infrared nanoscopy in aqueous has been a challenge.

PFIR fluid phase
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The action-based PFIR microscopy has several advantages in fluid operation:

  • Spatial resolution is not limited by the diffraction limit
  • Mechanical detection of photothermal effect bypasses the transmission attenuation after light-matter interactions
  • The peak force tapping mode has been proven to operate well in the fluid phase
  • Time-gated signal detection scheme is not drastically affected by the reduction of the cantilever quality factor

All these advantages make PFIR a promising method for aqueous phase nano-IR microscopy.

Dual-Color PFIR Microscopy

One possible extension of the PFIR-type photothermal detection mechanism is to incorporate multiple laser radiations with different beam characteristics (i.e., frequency, timing etc.) Such integration of multiple frequency excitations enables new capabilities for peak force photothermal microscopy.

Dual-color PFIR experimental schematic
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Experimental apparatus with two QCL sources

Dual-color PFIR microscopy is a variation of PFIR microscopy, in which two IR radiations of different frequencies are utilized to simultaneously deliver two PFIR images at different frequencies. The dual-color PFIR microscopy is good to image two chemical components of interest of non-sparse distributions, or with an open-loop AFM scanner, like the PeakForce Tapping enabled Multimode AFM of Bruker.

The dual-color PFIR microscopy is assembled with two separate mid infrared light sources (Quantum cascade laser QCL). The timing of the laser emission are synchronized to subsequent peak force tapping cycles. The photothermal responses are generated separately for each laser radiations and mechanically extracted with AFM cantilever oscillations.

Two signals from two IR excitations are simultaneously registered, together with the AFM topography and mechanical property channels.

The construction of dual-color PFIR microscope is described in our paper Analytical Chemistry, 94, 2, 1425-1431 (2022).

Dual-color PFIR images of E. coli cell surface
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Dual-color PFIR images of E. coli cell surface
Adhesion and PFIR spectra from E. coli surface
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PFIR spectra were collected from different locations of the E coli surface. These variations indicate heterogeneous chemical distribution on the cell surface.

Applications of PFIR

We are actively seeking collaborations to apply PFIR microscopy. If you have a sample that requires nanoscale multimodal characterization, feel free to reach out to Dr. Xu.

Aerosols are tiny particles levitating in the air. Small aerosol particles (PM2.5, diameter < 2.5 microns) are difficult to be filtered out and can reach respiratory trait. Small aerosol particles also float in the air for many hours, even days. The main transmission route for COVID-19 pathogen is believed to be aerosols. The PFIR microscopy provides a route for both chemical and mechanical characterizations under ambient conditions.

PFIR analysis of urban and indoor aerosol particles
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Urban aerosols
PFIR analysis of urban and indoor aerosol particles
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Indoor aerosols

Key Features

  • Simultaneous chemical and mechanical imaging
  • ~6nm spatial resolution, far beyond diffraction limit
  • Works in both air and liquid/aqueous phase
  • Compatible with soft polymers and biological materials
  • Multi-pulse mode for enhanced signal quality
  • Multimodal capability with KPFM integration

Applications

  • Aerosol particle analysis (PM2.5)
  • Biological cell characterization
  • Amyloid fibril protein studies
  • Oil shale and kerogen mapping
  • Phonon polaritons in h-BN
  • Click chemistry in aqueous phase

Interested in this research?

Contact us to learn more about collaboration opportunities.

Get in Touch
Analyst (2023)

Lock-in Amplifier Based Peak Force Infrared Microscopy

Andrea Dorsa, Qing Xie, Martin Wagner, and Xiaoji G. Xu

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Langmuir (2023)

What do different Modes of AFM-IR Mean for Measuring Soft Matter Surfaces?

Qing Xie and Xiaoji G. Xu

View Publication →