AFM-RAMAN-SICM System for Biological and Local Electrochemical Studies - NTEGRA MARLIN

The NTEGRA MARLIN available from NT-MDT Spectrum Instruments is a state-of-the-art AFM-RAMAN-SICM System used for local electrochemical and biological studies.

Key Features

  • QNM — power of SICM and AFM quantitative nanomechanical studies
  • AFM-SICM-Raman — triple synergy of robust nanocharacterization methods
  • Hopping mode ion conductance microscopy — non-contact imaging of jelly surfaces and living cells
  • Scanning electrochemical microscopy — electrochemistry at the end of the tip
  • Smart Patch Clamp — automatic nanopipette targeting for ion channel studies
  • Nano-injection — possibility to use nanopipettes with SICM feedback control for local sub-picoliter injections to single cells
  • High-speed, long-range high-resolution mapping at nanoscale

SICM Principle

Scanning Ion Conductance Microscopy (SICM) is an SPM method that involves using a nano-pipette (a sharp glass electrode) for non-contact 3D surface mapping at high resolution. In SICM, the distance between the probe and sample is regulated by reducing the ionic current that flows through the tip, as it comes close to the surface of the sample.

(a) Illustration of a scanning nanopipette probe operating in continuous scan mode colliding with a spherical object possessing a steep vertical slope. (b) Illustration of the hopping mode used in HPICM showing how the pipette is withdrawn to a position well above the sample before approaching the surface. (c,d) Topographical images of the same fixed hippocampal neuron obtained first with hopping mode (d) and then with continuous left-to-right raster scan mode (c), using the same nanopipette. Hopping mode algorithm applied to SICM allows to image uneven and convoluted samples at high resolution ensuring that pipette always approaches from above rather than “dragging” along the surface.

(a) Illustration of a scanning nanopipette probe operating in continuous scan mode colliding with a spherical object possessing a steep vertical slope. (b) Illustration of the hopping mode used in HPICM showing how the pipette is withdrawn to a position well above the sample before approaching the surface. (c,d) Topographical images of the same fixed hippocampal neuron obtained first with hopping mode (d) and then with continuous left-to-right raster scan mode (c), using the same nanopipette. Hopping mode algorithm applied to SICM allows to image uneven and convoluted samples at high resolution ensuring that pipette always approaches from above rather than “dragging” along the surface. Image Credit: Nature Meth. (2009) 6: 279-281.

Recording of the Webinar Presented by Prof. Yuri Korchev (Imperial College London): “NTEGRA Marlin: Bringing SICM to Users Research”

SICM Imaging of Living Cells

The non-contact algorithm of hopping SICM allows rapid, steady, and high-resolution imaging of soft and highly corrugated objects such as living cells under physiologically appropriate conditions.

The scanning technique guarantees that the probe always reaches the sample in a vertical direction, thereby making it possible to observe even those objects that are “suspended” in space.

Neuron from mouse hippocamp 10 × 10 × 6.3 μm.

Neuron from mouse hippocamp 10 × 10 × 6.3 μm

B16 melanoma cells 25 × 25 × 5.4 μm.

B16 melanoma cells 25 × 25 × 5.4 μm

PC3 human prostatic carcinoma cells 40 × 40 × 6.8 μm.

PC3 human prostatic carcinoma cells 40 × 40 × 6.8 μm

SICM image of live neuron from mouse hippocamp 40 × 40 × 13.3 μm.

SICM image of live neuron from mouse hippocamp 40 × 40 × 13.3 μm

Smart Patch Clamp

The smart patch-clamp integrates SICM and traditional patch-clamp. SICM produces a high-resolution topography, and subsequently ion current recording in a particular location.

Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels.

Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels. Image Credit: Neuron (2013) 79. 1067-77.

Extracellular pH Mapping of Single Living Cells

It is possible to implement extracellular pH mapping of living cancer cells with sensitivity and high spatial resolution through SICM by making use of a double-barrel nanopipette. pH-map and morphology of low-buffered living melanoma cells are displayed from the left. Scale bars show 20 μm.

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Image Credit: Nature Comm. (2019) 10, 5610.

SECM

Scanning Electrochemical Conductance Microscopy (SECM) is a robust tool for performing local electrochemical studies. It can be effectively used to image the dynamics of electron transport in battery electrodes or 2D systems.

Simultaneous SECCM topography (left) and current (right) images of a LiFePO4 electrode.

Simultaneous SECCM topography (left) and current (right) images of a LiFePO4 electrode. Image Credit: Nature Comm. (2014) 5, 5450.

High-Resolution AFM

High-resolution AFM microscopy comes in tapping, contact, and HybriD modes and is driven by the lowest signal-to-noise ratio of the OBD loop on the market, down to 25 FM/√Hz.

200 × 200 × 100 nm high-resolution AFM topography of rhinovirus particles.

200 × 200 × 100 nm high-resolution AFM topography of rhinovirus particles.

Quantitative Nanomechanical Studies (QNM)

The combination of HybriD Mode™ AFM and SICM extends the limits of real-time quantitative nanomechanical mapping to 10 orders of elastic modulus while maintaining the feasibility of single-point force spectroscopy experiments.

Non-invasive or less-invasive nature of tip-sample interaction enables the investigation of sensitive jelly and biological samples that are connected weakly to the substrate.

SICM E-Modulus map of live fibroblast cell. SICM E-modulus map of live fibroblast 25 × 25 μm, E = 2 Pa to 3.4 MPa.

SICM E-Modulus map of live fibroblast cell. SICM E-modulus map of live fibroblast 25 × 25 μm, E = 2 Pa to 3.4 MPa.

Streptavidin-biotin affinity single-molecule detection.

Streptavidin-biotin affinity single-molecule detection.

HybriD AFM E-modulus map of stem cell in HybriD Mode 18 × 18 μm, E = 200 kPa to 50 MPa.

HybriD AFM E-modulus map of stem cell in HybriD Mode 18 × 18 μm, E = 200 kPa to 50 MPa.

HybriD Mode™.

HybriD Mode™

Correlative Imaging

With the combination of flawless software and hardware of SICM/AFM and confocal fluorescence/Raman microscopy, an extensive range of additional data regarding the sample can be obtained.

Simultaneously measured SICM/AFM and fluorescence/Raman maps of precisely the same sample area provide additional data about chemical composition and physical properties of samples.

When provided with uniquely made probes that function as “nanoantennas”, the combination of SICM/AFM and Raman enables users to carry out optical mapping with a resolution less than diffraction limit and is known as Tip Enhanced Raman Scattering, or TERS.

Combined AFM-Raman microscopy studies of cyanobacterial film. AFM Phase map (a). Raman map showing the intensity distribution of the Raman band at 1521 cm−1 corresponding to beta-carotene (c). Raman-AFM overlay (b). Typical Raman spectrum of the sample containing a band at 520 cm−1 that is the Si-Si stretching mode of the silicon AFM tip and bands at 1160 cm−1 and 1521 cm−1assignable to beta-carotene.

Combined AFM-Raman microscopy studies of cyanobacterial film. AFM Phase map (a). Raman map showing the intensity distribution of the Raman band at 1521 cm−1 corresponding to beta-carotene (c). Raman-AFM overlay (b). Typical Raman spectrum of the sample containing a band at 520 cm−1 that is the Si-Si stretching mode of the silicon AFM tip and bands at 1160 cm−1 and 1521 cm−1assignable to beta-carotene.

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