Nanopore Reader (new!)

The new Nanopore Reader is a compact and portable read-out device with miniaturized flow cell for ultra low noise recordings of solid state nanopores and biological pores. The unit can be used for experimental activities such as resistive pulses, single molecule detection, DNA translocation. Thanks to its dimension and high data resolution, this unit is a versatile platform suitable for different applications if combined with the specific nanosensor, from water quality monitoring, environmental pollution analysis, agriculture tests and disease marker detection.

The Nanopore Reader uses two different types of flow cell depending on the nanopore typology.

For solid state nanopores, the flow cell measures only 15x25x8 mm, it integrates the Ag/AgCl electrodes, the nanopore slot and two compartments for fluids (30 µl bottom, 60 µl top).

5×5 mm squared or 3 mm round nanopore chip can be hosted into the flow cell.

For biological pores, the flow cell (BLMchip) measures 15x25x6 mm and integrates the Ag/AgCl electrodes. The septum is made by a Polyimide layer 50 µm thick and three micro hole sizes are available (100 µm, 150 µm, 200 µm). The BLMchip allows an easy access to the buffer solution of both the chambers by pipetting. The top chamber volume is 60 µl while the bottom chamber volume is 30 µl.

Applications

Exemplary current trace of KCVnts channel recorded at -60mV in symmetrical 1M KCl, PH 7 (with 1,25 KHz sampling rate and 625 Hz bandwidth). The channel protein was translated in vitro into NDs (nanodiscs) with DMPC membranes. The purified NDs in dilution with imidazole were directly administered to the bilayer formed by painting DPhPc over our 150 µm diameter BLMchip designed for biological pores. The open and closed level of the channel are indicated. The top panel shows the signal of the membrane at -60mV before the insertion of the channel protein. KCVnts is gift from Prof. G. Thiel (TU-Darmstadt University).
Exemplary current trace of a single α-Hemolysin channel insertion recorded at 5 KHz sampling rate (and 2,5 KHz bandwidth), applying a membrane potential of 100mV, in symmetrical 1M KCl, PH 7. PEG1000 molecules (already present in solution) translocate through the α-Hemolysin channel giving rise to negative individual blockades. The channel protein was directly added in solution and auto-assembled into the bilayers made of DPhPc.
A bilayer membrane was formed by painting DPHPC lipids (10mg/ml in n-octane) over the 100uM hole of the BLM-chip filled with asymmetrical HCl solutions. When a stable membrane was obtained, a small amount of gramicidin was added to the recording solutions. After some minutes, the formation of gramicidin dimers allowed the flow of H+ ions across the membrane.
Representative single channel currents of wt KcsA reconstituted in liposomes and incorporated in POPE:POPG lipid bilayers (w:w, 3:1). Data courtesy of Prof. M. F. Tsai (University of Colorado School of Medicine, USA).
Representative current trace showing single channel activity of chloride intracellular channel 1 (CLIC1) reconstituted in DPhPC lipid bilayer membranes painted over a 150 µm hole. Data courtesy of Prof. M. Mazzanti (University of Milan, Italy).
dsDNA fragment translocation data obtained with a 17-nm-diameter SiN x  pore at +200 mV, 1M KCl (10 mM Tris buffer, 1 mM EDTA and pH 8.0) for (a) 15 kbp, (b) 1000 bp, and (c) 400 bp dsDNA, and corresponding event duration histograms measured at 100 kHz bandwidth. Red curves are exponential fits to obtain the characteristic dwell times. Modified from Niedzwiecki et al Rev Sci Instrum. 2020 Mar 1;91(3):031301
350 nm sized polystyrene nanoparticle translocation obtained at 420 mV with a laser drilled 1µm pore in Polyimide layer. Data were acquired at 50 kHz SR, 25 kHz bandwidth.

How to perform a typical Lipid Bilayer experiment using the Nanopore Reader and the BLMchip

Features

  • Open input (RMS) noise (Voltage range ±700mV) : 70 fA rms @ 1kHz – 244 fA rms @ 10 kHz – 2,29 pA rms @ 100 kHz
  • Open input (RMS) noise (Voltage range ±2000mV) : 100 fA rms @ 1kHz – 415 fA rms @ 10 kHz – 3,51 pA rms @ 100 kHz
  • Current ranges: ±200pA (Gain 2.25GΩ), ±2nA (Gain 225MΩ), ±20nA (Gain 22.5MΩ), ±200nA (Gain 2.25MΩ)
  • Voltage hold ranges: ±700mV (ultra low noise); ±2000mV (low noise)
  • Parametric voltage protocols
  • Max sampling rate: 200 ksps
  • Selectable x4 oversampling (max final sampling rate 800 ksps)
  • Available bandwidth between 62.5 Hz to 100 kHz
  • Auto electrodes voltage offset fine compensation
  • Continuous Capacitance and Resistance estimation
  • USB powered
  • Size & Weight: 101 x 44 x 18 mm, 140 g
Click here to watch a video-description of the technical specification of eNPR amplifier and EDR3 software together with some representative applications and data