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Scanning ion conductance microscopy (SICM) is a type of in situ measurement technology for noncontact detection of samples in electrolytes with nanoscale resolution and has been used increasingly in biomedical and electrochemical fields in recent years. However, there is an inherent contradiction in the technique that makes SICM's sensitivity and accuracy difficult to balance. Higher sensitivity allows for faster probe speeds and higher scanning reliability but leads to lower accuracy, and vice versa. To resolve this problem, an adaptive sensitivity scanning method is proposed here that is designed to increase SICM's imaging efficiency without reducing its scanning reliability and accuracy. In the proposed scanning method, the sensitivity is automatically switched via the bias voltage based on the probe-sample distance. When the probe is located far away from the sample, the probe then predetects the sample position rapidly with high sensitivity. When the sample has been sensed in the high-sensitivity phase, the probe then detects the sample with low sensitivity. The basic theory and the feasibility of the alterable sensitivity detection strategy is also studied using the finite element method (FEM) and by performing experiments in this work. Finally, through testing of the standard silicon and polydimethylsiloxane (PDMS) samples, the proposed method is shown to increase SICM imaging efficiency significantly by up to 5 times relative to the conventional hopping mode without sacrificing the scanning accuracy and reliability.

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RESUMEN

Scanning ion conductance microscopy (SICM) as an emerging non-contact scanning probe microscopy technique and featuring its strong in-situ detectability for soft and viscous samples, is increasingly used in biomedical and materials related studies. In SICM measurements, employing theta pipette as SICM probe to scan sample is an effective method to extend the applications of SICM for multi-parameter measurement. There are two crucial but still unclear issues that influence the reliability and accuracy of the usage of theta pipette in the SICM measurements, which are the safe feedback threshold and the horizontal measurement offset. In this work, aiming at the theta pipette configuration of SICM, we systematically investigated the two issues of the theta pipette by both finite element method (FEM) simulation and SICM experiments. The FEM analysis results show that the safe feedback threshold of the one side barrel of the theta pipette is above 99.5%, and the horizontal measurement offset is ~0.53 times of the inner radius of the probe tip. Based on this, we proposed an improved scanning method used by the theta pipette to solve the reliability and accuracy problems caused by the feedback threshold too close to the reference current (100%) and the measurement offset error at the tip radius level. Then through testing the polydimethylsiloxane (PDMS) samples with different embossed patterns with the improved method of SICM, we can conclude that the improved method can enhance the scanning reliability by adding the double barrels approaching process and increase the positioning accuracy by compensating an offset distance. The theoretical analysis and the improved scanning method in this work demonstrate more property and usage details of the theta pipette, and further improve the reliability and accuracy of the diversified multifunctional applications of the theta pipette for SICM to meet the increasingly complex and precise research needs.

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Scanning ion conductance microscopy (SICM), as an emerging non-contact in situ topography measurement tool with nano resolution, has been increasingly used in recent years in biomedicine, electrochemistry and materials science. In the conventional measurement method of SICM, the sample topography is constructed according to the position of the probe at the feedback threshold of the ion current. Nevertheless, for different structures, a constant threshold cannot maintain a constant probe-sample distance. This phenomenon makes the measurement topography inconsistent with the real sample surface. In order to solve this problem and improve the measurement accuracy of SICM, a new ion conductance imaging method based on the approach curve spectrum is proposed in this work. In the new method, the local feature around the measurement point is firstly evaluated according to the change rate of ion current. Secondly, based on the local feature, the corresponding approach curve is searched from the prior approach curve spectrum to accurately evaluate the distance between the probe and the sample. Finally, the sample topography is constructed by the probe position subtracting the probe-sample distance. In this paper, we verify the feasibility of the new imaging method by combining finite element theory and experiments. To examine the measurement accuracy, the standard strip silicon and cylindrical polydimethylsiloxane (PDMS) samples are tested, and the improved imaging method can obtain the topography closer to the real samples and reduce the volumetric measurement error by 5.4%. The implementation of the new imaging method will further promote SICM application in related research fields.

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Scanning ion conductance microscopy (SICM), one kind of scanning probe microscopy technique, featuring the advantage of non-contact imaging of sample surfaces in three dimensions with high resolution, has been widely applied in characterizations of sample topography, especially for soft materials. However, the time consuming imaging process of SICM restricts its further applications, such as in characterization of dynamic change of sample surface. In this work, a fast control mode of SICM, named as a continuous control mode, has been developed. In this mode, the SICM probe (i.e., pipette) is controlled by speed instructions in the axial direction of pipette (Z axis), and the pipette position is determined by the position sensor. Compared to the conventional piezo control mode of SICM (i.e., the stepwise control mode), in which the pipette is controlled by the position instructions and moves step by step, the continuous control mode can perform the continuous movement of the pipette in Z axis and overcome the time consuming problem caused by the repeated acceleration and deceleration of the pipette during the stepwise mode. Moreover, the imaging resolution in Z axis is not restricted by the pipette movement step and the imaging rate in the continuous control mode can be significantly enhanced without losing imaging quality. The approach speed of pipette in the continuous control mode can reach at 300 nm/ms, which is much faster than that in the stepwise mode. The surfaces of the soft polydimethylsiloxane (PDMS) samples with three different patterns, the hard metal grating sample and the cardiac fibroblasts as the biological sample demo were comparably scanned by SICM using the continuous control mode and the stepwise approach mode, respectively. The obtained SICM images of the sample topography prove that the continuous control mode can not only reduce the imaging deviation, but also efficiently improve the scanning rate of SICM. Furthermore, the continuous control mode can reconstruct the sample topography more stably compared to the stepwise control mode. The continuous control mode developed in this work can provide an efficient and reliable control strategy for improving the imaging performance of SICM system, and therefore can be potentially applied in dynamic characterizations of various samples in material science, biology and chemistry fields.

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Scanning ion conductance microscopy (SICM) is a non-contact surface topography measurement technique that has been increasingly used for soft surfaces such as living biological samples. An approach-retract scanning (ARS) mode is widely used to avoid collision between the SICM probe (i.e., pipette) and an abrupt increase in sample profile. However, the redundant pipette trajectory in the ARS mode lengthens the scan time, thus reducing SICM efficiency and time resolution. To avoid this problem, a new scanning mode is discussed that adds horizontal movement at each measurement point to predict the upcoming sample topography via variation in ion current. The pipette then retracts in response to raised topography, while it raster scans flat or downhill topography. The feasibility was verified by finite element analysis and experimental tests on three kinds of soft samples: polydimethylsiloxane, mice cardiac fibroblasts, and breast cancer cells. The pixel detection frequency during imaging and the mean square error of the sample topography were compared for the two modes. The new scanning mode enhances the SICM imaging rate without loss of imaging quality or scanning stability, while it increases efficiency and time resolution. It thus has an improved performance for characterizing biological samples.

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In this paper, a new double micropipettes configuration mode of scanning ion conductance microscopy (SICM) is presented to better overcome ionic current drift and further improve the performance of SICM, which is based on a balance bridge circuit. The article verifies the feasibility of this new configuration mode from theoretical and experimental analyses, respectively, and compares the quality of scanning images in the conventional single micropipette configuration mode and the new double micropipettes configuration mode. The experimental results show that the double micropipettes configuration mode of SICM has better effect on restraining ionic current drift and better performance of imaging. Therefore, this article not only proposes a new direction of overcoming the ionic current drift but also develops a new method of SICM stable imaging.

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