A small scanning tunnelling microscope with large scan range for biological studies

We have developed a scanning tunnelling microscope specially designed for biological applications presenting some new features: the scanner tube is mounted parallel to the surface of the sample which enables a high resolution optical microscope to be brought close to the sample when working in air or liquids. The maximum scan range is 5×20 μm with a vertical range of 20 μm and the total size of the system does not exceed 10×40 mm. The piezo-sensitivity of the scanner tube versus applied voltage was analysed by interferometry measurements and by using scanning tunnelling microscopes. We found a value for the piezoelectric constant d₁₃ of ?1·71 Å/V at low voltages (under a few volts) going up to ?2 Å/V for higher voltages. Large-scale images of a carbon grid showed a surprisingly good linearity of the scanner tube.     

石英音叉扫描探针显微镜

石英音叉是一种谐振频率稳定、品质因数高的时基器件,其音叉臂的谐振参数(谐振振幅和谐振频率)对微力极其敏感。利用石英音叉对外力的敏感性,与钨探针结合,构成一种新型的表面形貌扫描测头。该测头与xyz压电工作台结合,利用测头音叉臂谐振频率对扫描微力的敏感性,研制基于相位反馈控制的扫描探针显微镜。首先介绍石英音叉测头的构成、工作原理和特性测试,以及由该测头构建的扫描探针显微镜的结构和测试、分析。通过对测头和系统的测试结果分析,系统达到1.2 nm的垂直分辨率,并通过对一维栅的测量,给出扫描获得的试样表面微观形貌图以及相位图,证明系统的有效性。另外,由于采用大长径比的钨探针,该系统具有测量大深宽比微器件表面轮廓的能力。     

A scanner for an ultrahigh-vacuum low-temperature scanning tunneling microscope

A scanner for an ultrahigh-vacuum low-temperature scanning tunneling microscope is described. It has a high resonance frequency (>30 kHz) and a small thermal-drift rate (≤1 nm/°C) at room temperature. The scanner feeds the tip to the sample at a distance of up to 3 mm and positions it in the sample plane on a 4 × 4-mm area. These characteristics of the scanner allow one to study atomic structures at temperature variations from 5 to 300 K with objects under study remaining in view of the microscope. The scanner has a horizontal attachment for a sample with a size of up to 6 × 6 × 3mm and ensures a scanning field of 4.8 × 4.8 × 0.6 μm at 300 K and 0.8 × 0.8 × 0.1 μm at 5 K, as well as the possibility of heating to 150°C and easily replacing the sample and tip with vacuum manipulators.     

Measurement of probe displacement to the thermal resolution limit in photonic force microscopy using a miniature quadrant photodetector

A photonic force microscope comprises of an optically trapped micro-probe and a position detection system to track the motion of the probe. Signal collection for motion detection is often carried out using the backscattered light off the probe-however, this mode has problems of low S/N due to the small backscattering cross sections of the micro-probes typically used. The position sensors often used in these cases are quadrant photodetectors. To ensure maximum sensitivity of such detectors, it would help if the detector size matched with the detection beam radius after the condenser lens (which for backscattered detection would be the trapping objective itself). To suit this condition, we have used a miniature displacement sensor whose dimensions makes it ideal to work with 1:1 images of micrometer-sized trapped probes in the backscattering detection mode. The detector is based on the quadrant photo-integrated chip in the optical pick-up head of a compact disc player. Using this detector, we measured absolute displacements of an optically trapped 1.1 μm probe with a resolution of ～10 nm for a bandwidth of 10 Hz at 95% significance without any sample or laser stabilization. We characterized our optical trap for different sized probes by measuring the power spectrum for each probe to 1% accuracy, and found that for 1.1 μm diameter probes, the noise in our position measurement matched the thermal resolution limit for averaging times up to 10 ms. We also achieved a linear response range of around 385 nm with cross talk between axes ?4% for 1.1 μm diameter probes. The detector has extremely high bandwidth (few MHz) and low optical power threshold-other factors that can lead to its widespread use in photonic force microscopy.     

A scanning near-field optical microscope having scanning electron tunnelling microscope capability using a single metallic probe tip

A new microscope system that has the combined capabilities of a scanning near-field optical microscope (SNOM) and a scanning tunnelling microscope (STM) is described. This is achieved with the use of a single metallic probe tip. The distance between the probe tip and the sample surface is regulated by keeping the tunnelling current constant. In this mode of operation, information about the optical properties of the sample, such as its refractive index distribution and absorption characteristics, can be disassociated from the information describing its surface structure. Details of the surface structure can be studied at resolutions smaller than the illumination wavelength. The performance of the microscope is evaluated by analysing a grating sample that was made by coating a glass substrate with gold. The results are then compared with the corresponding SNOM and STM images of the grating.     

原子力显微镜管式扫描器运动学建模与误差分析

针对样本扫描模式原子力显微镜,对其管式扫描器-样本-探针系统进行了运动学分析,建立了该系统的运动学模型,该模型表明:对于给定原子力显微镜扫描器,样本上与探针接触点的横向和纵向位移取决于探针尖端相对于扫描管轴心的偏置量、所加电压(或名义扫描范围)及样本厚度。据此模型,对由于弯曲运动模式所产生的两种重要误差—交叉耦合误差及扫描范围误差进行了定量分析,分析表明:扫描范围误差主要受样本厚度及名义扫描范围影响,而Z向交叉耦合误差主要受探针偏置量及名义扫描范围影响,实验验证了所建立的运动学模型和误差计算公式的正确性;另外,还提出了相应的减小误差的方法。     

X轴分离式高速原子力显微镜系统设计

为了提高原子力显微镜(Atomic Force Microscope,AFM)的成像速度,本文提出了一种新的AFM结构设计方案并搭建了相应的实验系统。在该方案中,Y、Z扫描器集成于测头内驱动探针进行慢轴扫描和形貌反馈;X扫描器与测头分离,驱动样品做快轴扫描。X扫描器采用高刚性的独立一维纳米位移台,能够承载尺寸和质量较大的样品高速往复运动而不易发生共振;同时Z扫描器的载荷实现最小化,固有频率得以显著提高。为了避免测头的扫描运动引起检测光束与探针相对位置的偏差,设计了一种随动式光杠杆光路;为了便于装卸探针以及精确调整激光在探针上的反射位置,设计了基于磁力的探针固定装置和相应的光路调节方案。对所搭建的AFM系统的初步测试结果表明,该系统在采用三角波驱动和简单PID控制算法的情况下,可搭载尺寸达数厘米且质量超过10g的较大样品实现13μm×13μm范围50Hz行频的高速成像。     

Rapid measurement of a high step microstructure with 90° steep sidewall

A prototype STM system with high aspect ratio measurement capability is developed to fulfill accurate profile measurement of a high step microstructure with 90° steep sidewall. Distinguished from the traditional STM, the new system consists of a long range piezoelectric (PZT) actuator with full stroke of 60 μm as Z-direction servo scanner, a specially customized high aspect ratio STM probe with effective tip length of 300 μm, and an X-Y motorized driven stage for planar scanning. A tilt stage is used to adjust the probe-sample relative angle to compensate the evitable non-parallel effects. Based on the new STM system, sample-tilt-scanning methodology is proposed for eliminating the scanning blind region between the probe and the microstructure. A high step microstructure with height of 23 μm, 90° steep sidewall and width of 50μm has been successfully measured. The slope angle of the sidewall has been achieved to be 85° and the step height at the rising edge and the trench depth at the falling edge are both measured to be 22.96 μm. The whole measuring process only spent less than 10 min. It provides an effective and nondestructive solution for the measurement of high step or deep trench microstructures. In addition, this work also opens the way for further study on sidewall roughness and the tip-sample interaction at the edge of the sidewall, which are highly valuable for fabrication and quality control of high step microstructures.     

Compact ultra-fast vertical nanopositioner for improving scanning probe microscope scan speed

The mechanical design of a high-bandwidth, short-range vertical positioning stage is described for integration with a commercial scanning probe microscope (SPM) for dual-stage actuation to significantly improve scanning performance. The vertical motion of the sample platform is driven by a stiff and compact piezo-stack actuator and guided by a novel circular flexure to minimize undesirable mechanical resonances that can limit the performance of the vertical feedback control loop. Finite element analysis is performed to study the key issues that affect performance. To relax the need for properly securing the stage to a working surface, such as a laboratory workbench, an inertial cancellation scheme is utilized. The measured dominant unloaded mechanical resonance of a prototype stage is above 150 kHz and the travel range is approximately 1.56 μm. The high-bandwidth stage is experimentally evaluated with a basic commercial SPM, and results show over 25-times improvement in the scanning performance.     

A confocal laser microscope scanner for digital recording of optical serial sections

A confocal laser microscope scanner developed at our institute is described. Since an ordinary microscope is used, it is easy to view the specimen prior to scanning. Confocal imaging is obtained by laser spot illumination, and by focusing the reflected or fluorescent light from the specimen onto a pinhole aperture in front of the detector (a photomultiplier tube). Two rotating mirrors are used to scan the laser beam in a raster pattern. The scanner is controlled by a microprocessor which coordinates scanning, data display, and data transfer to a host computer equipped with an array processor. Digital images with up to 1024 × 1024 pixels and 256 grey levels can be recorded. The optical sectioning property of confocal scanning is used to record thin (～ 1 μm) sections of a specimen without the need for mechanical sectioning. By using computer-control to adjust the focus of the microscope, a stack of consecutive sections can be automatically recorded. A computer is then used to display the 3-D structure of the specimen. It is also possible to obtain quantitative information, both geometric and photometric. In addition to confocal laser scanning, it is easy to perform non-confocal laser scanning, or to use conventional microscopic illumination techniques for (non-confocal) scanning. The design has proved reliable and stable, requiring very few adjustments and realignments. Results obtained with this scanner are reported, and some limitations of the technique are discussed.     

A fast and versatile scan unit for scanning probe microscopy

In scanning probe microscopy (SPM), the image acquisition time is usually very long because of the limited speed with which the scanning device can trace the topography of the specimen under feed-back control. This limitation is often brought about by the natural frequency of the scanner in the direction perpendicular to the sample plane that confines the usable bandwidth of the feed-back loop. In this paper, we present a piezo-ceramic scanner that provides a large scan range and at the same time allows for adjustment of the probe-to-sample distance faster by about one order of a magnitude than a conventional setup. This is achieved through the combination of a large single tube scanner that provides a high-scan range and a small piezo element for swift motion in the direction perpendicular to the sample plane. The natural frequency in this direction lies at about 275 kHz. We outline the design considerations to avoid disturbing excitation of the scanner through the fast piezo element.     

A versatile multipurpose scanning probe microscope

A combined scanning probe microscope has been developed that allows simultaneous operation as a non‐contact/tapping mode atomic force microscope, a scattering near‐field optical microscope, and a scanning tunnelling microscope on conductive samples. The instrument is based on a commercial optical microscope. It operates with etched tungsten tips and exploits a tuning fork detection system for tip/sample distance control. The system has been tested on a p‐doped silicon substrate with aluminium depositions, being able to discriminate the two materials by the electrical and optical images with a lateral resolution of 130 nm.     

A 2D MEMS mirror with sidewall electrodes applied for confocal MACROscope imaging

This paper presents microelectromechanical system micromirrors with sidewall electrodes applied for use as a Confocal MACROscope for biomedical imaging. The MACROscope is a fluorescence and brightfield confocal laser scanning microscope with a very large field of view. In this paper, a microelectromechanical system mirror with sidewall electrodes replaces the galvo-scanner and XYZ-stage to improve the confocal MACROscope design and obtain an image. Two micromirror-based optical configurations are developed and tested to optimize the optical design through scanning angle, field of view and numerical aperture improvement. Meanwhile, the scanning frequency and control waveform of the micromirror are tested. Analysing the scan frequency and waveform becomes a key factor to optimize the micromirror-based confocal MACROscope. When the micromirror is integrated into the MACROscope and works at 40 Hz, the micromirror with open-loop control possesses good repeatability, so that the synchronization among the scanner, XYZ-stage and image acquisition can be realized. A laser scanning microscope system based on the micromirror with 2 μm width torsion bars was built and a 2D image was obtained as well. This work forms the experimental basis for building a practical confocal MACROscope.     

Wide-band acousto-optic deflectors for large field of view two-photon microscope

Acousto-optic deflector (AOD) is an attractive scanner for two-photon microscopy because it can provide fast and versatile laser scanning and does not involve any mechanical movements. However, due to the small scan range of available AOD, the field of view (FOV) of the AOD-based microscope is typically smaller than that of the conventional galvanometer-based microscope. Here, we developed a novel wide-band AOD to enlarge the scan angle. Considering the maximum acceptable acoustic attenuation in the acousto-optic crystal, relatively lower operating frequencies and moderate aperture were adopted. The custom AOD was able to provide 60 MHz 3-dB bandwidth and 80% peak diffraction efficiency at 840 nm wavelength. Based on a pair of such AOD, a large FOV two-photon microscope was built with a FOV up to 418.5 μm (40× objective). The spatiotemporal dispersion was compensated simultaneously with a single custom-made prism. By means of dynamic power modulation, the variation of laser intensity within the FOV was reduced below 5%. The lateral and axial resolution of the system were 0.58-2.12 μm and 2.17-3.07 μm, respectively. Pollen grain images acquired by this system were presented to demonstrate the imaging capability at different positions across the entire FOV.     

Construction and testing of a nanomachining instrument

This paper presents a nanomachining instrument that was developed for conducting nanocutting, nanoscratching, and nanoindentation experiments. A piezoelectric tube scanner (PZT) is employed to generate three-dimensional machining motions. The sample is moved by the PZT, and the tool is kept stationary during machining. The machining forces are measured by force sensors with a resolution of sub-milliNewtons. The instrument is compact and can be used inside optical microscopes and scanning electron microscopes. In this paper, depth-sensing indentation experiments were performed to test the basic performance of the instrument. The indentation displacement was measured by a capacitance probe situated inside the PZT tube. An experimental system was constructed to locate and image indentations. The system consists of a high magnification microscope to measure coordinates of the indentation relative to a reference corner point on the sample, and an AFM equipped with an on-axis optical imaging system for locating the indentation. A technique was also employed to establish the tool-sample contact to nanometer accuracy. Indentation experiments were carried out on three kinds of materials with different hardness. Experimental results demonstrated the instrument has the ability of performing depth-sensing indentations. The frame compliance was also evaluated from the indentation results.     

Combining optical tweezers and scanning probe microscopy to study DNA-protein interactions

We present the first results obtained with a new instrument designed and built to study DNA-protein interactions at the single molecule level. This microscope combines optical tweezers with scanning probe microscopy and allows us to locate DNA-binding proteins on a single suspended DNA molecule. A single DNA molecule is stretched taut using the optical tweezers, while a probe is scanned along the molecule. Interaction forces between the probe and the sample are measured with the optical tweezers. The instrument thus enables us to correlate mechanical and functional properties of bound proteins with the tension within the DNA molecule. The typical friction force between a micropipette used as probe and a naked DNA molecule was found to be <1 pN. A 16 micro m DNA molecule with approximately 10-15 digoxygenin (DIG) molecules located over a 90 nm range in the middle of the DNA was used as a model system. By scanning with an antidigoxygenin (alpha-DIG) antibody-coated pipette we were able to localize these sites by exploiting the high binding affinity between this antibody-antigen pair. The estimated experimental resolution assuming an infinitesimally thin and rigid probe and a single alpha-DIG/DIG bond was 15 nm.     

Note: long range and accurate measurement of deep trench microstructures by a specialized scanning tunneling microscope

A compact but practical scanning tunneling microscope (STM) with high aspect ratio and high depth capability has been specially developed. Long range scanning mechanism with tilt-adjustment stage is adopted for the purpose of adjusting the probe-sample relative angle to compensate the non-parallel effects. A periodical trench microstructure with a pitch of 10 μm has been successfully imaged with a long scanning range up to 2.0 mm. More innovatively, a deep trench with depth and step height of 23.0 μm has also been successfully measured, and slope angle of the sidewall can approximately achieve 67°. The probe can continuously climb the high step and exploring the trench bottom without tip crashing. The new STM could perform long range measurement for the deep trench and high step surfaces without image distortion. It enables accurate measurement and quality control of periodical trench microstructures.     

An evanescent-field optical microscope

The classic diffraction limit of resolution in optical microscopy (～γ/2) can be overcome by detecting the diffracted field of a submicrometre-size probe in its near field. The present stage of this so-called scanning near-field optical microscopy (SNOM) is reviewed. An evanescent-field optical microscope (EFOM) is presented in which the near-field regime is provided by the exponentially decaying evanescent field caused by total internal reflection at a refractive-index transition. A sample placed in this field causes a spatial variation of the evanescent field which is characteristic for the dielectric and topographic properties of the sample. The evanescent field is frustrated by a dielectric probe and thus converted into a radiative field. In our case the probe consists either of an etched optical fibre or of a highly sharpened diamond tip. The probe is scanned over the sample surface with nanometre precision using a piezo-electric positioner. The distance between probe and sample is controlled by a feedback on the detected optical signal. The resolution of the microscope is determined by both the gradient of the evanescent field and the sharpness of the tip. Details of the experimental set-up are discussed. The coupling of the evanescent field to the submicrometre probe as a function of probe-sample distance, angle of incidence and polarization has been characterized quantitatively. The observed coupling is generally in agreement with presented theoretical calculations. Microscopy has been performed on a regular latex sphere structure, which clearly demonstrates the capacity of the evanescent-field optical microscope for nanometre-scale optical imaging. Resolution is typically 100 nm laterally and 10 nm vertically. The technique is promising for biological applications, especially if combined with optical spectroscopy.     

Near-field optical microscope working on TEM wave

A new type of probe for the near-field optical microscope incorporating metallic strips on the surface of a dielectric cone (microstrip probe) is proposed. The numerical simulation has shown a significant improvement in optical efficiency for this type of probe compared with the conventional probe. It was found theoretically that scanning near-field optics with the microstrip probe is promising for applications in optical information recording and can be employed as optical heating elements in magnetic information recording. Application of the microstrip probe in nonlinear optical research of nanoscaled medium is possible due to strong electric and magnetic field near the aperture and weak dependence of its transmission parameters on the frequency of the incident beam. A model for the near-field strip probe with cleaved apex is proposed. A method is presented to control the distance between the probe apex and sample surface based on mechanical vibrations of the cleaved apex excited by applying voltage pulses. The oscillation amplitude and attenuation can be determined by measuring the amplitude of microwave radiation excited by oscillations of charges with opposite sign at the probe apex. The investigation was based on mathematical models and experiments necessary to confirm the theoretical prediction.