High-speed atomic force microscope combined with single-molecule fluorescence microscope

Fukuda, Shingo; Uchihashi, Takayuki; Iino, Ryota; Okazaki, Yasutaka; Yoshida, Masato; Igarashi, Kiyohiko; Ando, Toshio

doi:10.1063/1.4813280

Cited by 70 publications

(60 citation statements)

References 47 publications

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“…The use of fluorescence-related microscopy can help us to discern the detailed subcellular structures (e.g., cytoskeletons, cellular nuclei, cytoplasm) during the AFM indenting; thus, it greatly complemented the indentation by AFM. In addition, researchers have successfully combined high-speed AFM with fluorescence microscopy in the recent researches [82][83][84]. Suzuki et al [82] combined high-speed AFM with an optical fluorescence microscope by developing a tip-scanning system, and the results (observing the dynamics of living cells) showed that this system can remarkably improve the time resolution.…”

Section: Challenge and Outlookmentioning

confidence: 99%

“…Suzuki et al [82] combined high-speed AFM with an optical fluorescence microscope by developing a tip-scanning system, and the results (observing the dynamics of living cells) showed that this system can remarkably improve the time resolution. Fukuda et al [83] have combined high-speed AFM with total internal reflection fluorescence microscopy, and this system can perform simultaneous topographic and fluorescence imaging at the single molecule level. Colom et al [84] have developed a hybrid high-speed atomic forceoptical microscope for visualizing single-membrane proteins on eukaryotic cells, allowing high-speed AFM imaging about 1,000 times faster than conventional AFM/ optical microscopy setups.…”

Section: Challenge and Outlookmentioning

confidence: 99%

“…In order to address these issues, researchers from different disciplines (e.g., physics, engineering, biology, information, and automation) need to work together. In recent years, researchers are continuing to enhance and expand the performance of AFM indentation technique, such as high-speed AFM [82][83][84], tomographic contact resonance AFM [88], and force curvebased AFM [25,89]. The further expanding application of AFM indentation in the life sciences will bring tremendous changes in biomechanics, clinical medicine, and drug development.…”

Section: Challenge and Outlookmentioning

confidence: 99%

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Research progress in quantifying the mechanical properties of single living cells using atomic force microscopy

Liu

et al. 2014

Chin. Sci. Bull.

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The advent of atomic force microscopy (AFM) provides a powerful tool for investigating the behaviors of single living cells under near physiological conditions. Besides acquiring the images of cellular ultra-microstructures with nanometer resolution, the most remarkable advances are achieved on the use of AFM indenting technique to quantify the mechanical properties of single living cells. By indenting single living cells with AFM tip, we can obtain the mechanical properties of cells and monitor their dynamic changes during the biological processes (e.g., after the stimulation of drugs). AFM indentation-based mechanical analysis of single cells provides a novel approach to characterize the behaviors of cells from the perspective of biomechanics, considerably complementing the traditional biological experimental methods. Now, AFM indentation technique has been widely used in the life sciences, yielding a large amount of novel information that is meaningful to our understanding of the underlying mechanisms that govern the cellular biological functions. Here, based on the authors' own researches on AFM measurement of cellular mechanical properties, the principle and method of AFM indentation technique was presented, the recent progress of measuring the cellular mechanical properties using AFM was summarized, and the challenges of AFM single-cell nanomechanical analysis were discussed.

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Section: Challenge and Outlookmentioning

confidence: 99%

Section: Challenge and Outlookmentioning

confidence: 99%

Section: Challenge and Outlookmentioning

confidence: 99%

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Research progress in quantifying the mechanical properties of single living cells using atomic force microscopy

Liu

et al. 2014

Chin. Sci. Bull.

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“…More seriously, the cantilever blocks the optical path of fluorescence microscopy. To overcome this problem, tip-scan HS-AFM has been developed, in which the laser beam of the OBD detector is steered with a two-dimensional mirror tilter to track the lateral motion of the cantilever (Fukuda et al 2013). This system has been combined with total internal reflection fluorescence microscopy.…”

Section: Hybrid Hs-afm/optical Microscopy Systemmentioning

confidence: 99%

High-speed atomic force microscopy and its future prospects

2017

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Various techniques have been developed and used to investigate how proteins produce complex biological architectures and phenomena. Among these techniques, high-speed atomic force microscopy (HS-AFM) holds a unique position. It is only HS-AFM that allows the simultaneous assessment of structure and dynamics of single protein molecules in action. This new microscopy tool has been successfully applied to a variety of proteins, from motor proteins to membrane proteins, antibodies, enzymes, and even to intrinsically disordered proteins. And yet there still remain many biomolecular phenomena that cannot be addressed by HS-AFM in its current form. Here, I present a brief history of HS-AFM development, describe the current state of HS-AFM, and then discuss which new biological scanning probe microscopy techniques will be coming up next.

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“…One of the common requirements of tip-scan AFM that employs the widely used optical lever method of detecting cantilever's deflection is that the laser beam focused onto the cantilever should be able to track the lateral motion of the cantilever. This tracking adoptable to tip-scan HS-AFM was recently accomplished by laser beam scan in synchrony with the cantilever's lateral motion using either a mirror tilter 20 or a focusing lens embedded in the XY-scanner. 21 The combined system of total internal reflection fluorescence microscopy (TIRFM) and tip-scan HS-AFM equipped with the mirror tilter was demonstrated to be able to capture topographic AFM and single-molecule fluorescent images simultaneously at a few frames/s (fps).…”

Section: Introductionmentioning

confidence: 99%

Method of mechanical holding of cantilever chip for tip-scan high-speed atomic force microscope

Fukuda

Uchihashi

Ando

2015

Review of Scientific Instruments

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Articles you may be interested in Note: High-speed Z tip scanner with screw cantilever holding mechanism for atomic-resolution atomic force microscopy in liquid Rev. Sci. Instrum. 85, 126106 (2014) In tip-scan atomic force microscopy (AFM) that scans a cantilever chip in the three dimensions, the chip body is held on the Z-scanner with a holder. However, this holding is not easy for high-speed (HS) AFM because the holder that should have a small mass has to be able to clamp the cantilever chip firmly without deteriorating the Z-scanner's fast performance, and because repeated exchange of cantilever chips should not damage the Z-scanner. This is one of the reasons that tip-scan HS-AFM has not been established, despite its advantages over sample stage-scan HS-AFM. Here, we present a novel method of cantilever chip holding which meets all conditions required for tip-scan HS-AFM. The superior performance of this novel chip holding mechanism is demonstrated by imaging of the α 3 β 3 subcomplex of F 1 -ATPase in dynamic action at ∼7 frames/s. C 2015 AIP Publishing LLC. [http://dx

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High-speed atomic force microscope combined with single-molecule fluorescence microscope

Cited by 70 publications

References 47 publications

Research progress in quantifying the mechanical properties of single living cells using atomic force microscopy

Research progress in quantifying the mechanical properties of single living cells using atomic force microscopy

High-speed atomic force microscopy and its future prospects

Method of mechanical holding of cantilever chip for tip-scan high-speed atomic force microscope

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