Changes in Plasma Membrane Surface Potential of PC12 Cells as Measured by Kelvin Probe Force Microscopy

Tsai, Chia Chang; Hung, Hui Hsing; Liu, Chien Pang; Chen, Yit‐Tsong; Pan, Chien‐Yuan

doi:10.1371/journal.pone.0033849

Cited by 26 publications

(28 citation statements)

References 35 publications

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“…FDA can permeate the intact cellular membrane freely, and then be decomposed as fluorescein (with polarity and fluorescence) by the lipase inside the cell. The length of the cell bodies was 9.8 ± 1.2 µm (N = 10), which corresponds with well with previous results (Jeon et al, 2010;Tsai et al, 2012;Zhao et al, 2019b). The green fluorescence can be induced by fluorescein.…”

Section: Resultssupporting

confidence: 92%

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Direct investigation of current transport in cells by conductive atomic force microscopy

Zhao

Cheong

et al. 2019

Journal of Microscopy

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Currents play critical roles in neurons. Direct observation of current flows in cells at nanometre dimensions and picoampere current resolution is still a daunting task. In this study, we investigated the current flows in hippocampal neurons, PC12 cells and astrocytes in response to voltages applied to the cell membranes using conductive atomic force microscopy (CAFM). The spines in the hippocampal neurons play crucial roles in nerve signal transfer. When the applied voltage was greater than 7.2 V, PC12 cells even show metallic nanowirelike characteristics. Both the cell body and glial filaments of astrocytes yielded CAFM test results that reflect different electrical conductance. To our best knowledge, the electrical characteristics and current transport through components of cells (especially neurons) in response to an applied external voltage have been revealed for the first time at nanometre dimensions and picoampere current levels. We believe that such studies will pave new ways to study and model the electrical characteristics and physiological behaviours in cells and other biological samples.

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Section: Resultssupporting

confidence: 92%

“…The cells were comprised by the cell bodies and processes. The length of the cell bodies was 9.8 ± 1.2 µm (N = 10), which corresponds with well with previous results (Jeon et al, 2010;Tsai et al, 2012;Zhao et al, 2019b). The processes (depicted by white arrows in Fig.…”

Section: Resultssupporting

confidence: 92%

Direct investigation of current transport in cells by conductive atomic force microscopy

Zhao

Cheong

et al. 2019

Journal of Microscopy

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“…Recently, Tsai et al . () showed that KPFM can be applied to analyze the physiological conditions of the plasma membrane in response to various stimulants.…”

Section: Methodsmentioning

confidence: 99%

A hemicellulose‐bound form of silicon inhibits cadmium ion uptake in rice (Oryza sativa) cells

Cai

et al. 2015

New Phytologist

285

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Summary Silicon (Si) alleviates cadmium (Cd) toxicity in rice (Oryza sativa). However, the chemical mechanisms at the single‐cell level are poorly understood. Here, a suspension of rice cells exposed to Cd and/or Si treatments was investigated using a combination of plant cell nutritional, molecular biological, and physical techniques including in situ noninvasive microtest technology (NMT), polymerase chain reaction (PCR), inductively coupled plasma mass spectroscopy (ICP‐MS), and atomic force microscopy (AFM) in Kelvin probe mode (KPFM). We found that Si‐accumulating cells had a significantly reduced net Cd2+ influx, compared with that in Si‐limited cells. PCR analyses of the expression levels of Cd and Si transporters in rice cells showed that, when the Si concentration in the medium was increased, expression of the Si transporter gene Low silicon rice 1 (Lsi1) was up‐regulated, whereas expression of the gene encoding the transporter involved in the transport of Cd, Natural resistance‐associated macrophage protein 5 (Nramp5), was down‐regulated. ICP‐MS results revealed that 64% of the total Si in the cell walls was bound to hemicellulose constituents following the fractionation of the cell walls, and consequently inhibited Cd uptake. Furthermore, AFM in KPFM demonstrated that the heterogeneity of the wall surface potential was higher in cells cultured in the presence of Si than in those cultured in its absence, and was homogenized after the addition of Cd. These results suggest that a hemicellulose‐bound form of Si with net negative charges is responsible for inhibition of Cd uptake in rice cells by a mechanism of [Si‐hemicellulose matrix]Cd complexation and subsequent co‐deposition.

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“…Negatively charged surfaces prevent the attachment of Bacillus thuringiensis spores to mica, silicon, and gold 6 . Changes in cellular membrane potential are indicative of changes in cellular attachment and motility 5,7 . It has been observed that electrically homogeneous surfaces more readily promote microbial adhesion 5 .…”

Section: Introductionmentioning

confidence: 99%

“…Samples do not necessarily have to be conductive (i.e., biological samples). Imaging can be carried out on mica, glass, and silicon surfaces (non-conductive) as long as the non-conductive surface is thin and there is an underlying conductive material 6,7 . The CPD is equivalent to the surface potential voltage and can be described as the difference in the work functions between the tip (φ tip ) and sample (φ sample ), divided by the negative electron charge (-e).…”

Section: Introductionmentioning

confidence: 99%

Surface Potential Measurement of Bacteria Using Kelvin Probe Force Microscopy

Birkenhauer

Neethirajan

2014

JoVE

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Surface potential is a commonly overlooked physical characteristic that plays a dominant role in the adhesion of microorganisms to substrate surfaces. Kelvin probe force microscopy (KPFM) is a module of atomic force microscopy (AFM) that measures the contact potential difference between surfaces at the nano-scale. The combination of KPFM with AFM allows for the simultaneous generation of surface potential and topographical maps of biological samples such as bacterial cells. Here, we employ KPFM to examine the effects of surface potential on microbial adhesion to medically relevant surfaces such as stainless steel and gold. Surface potential maps revealed differences in surface potential for microbial membranes on different material substrates. A step-height graph was generated to show the difference in surface potential at a boundary area between the substrate surface and microorganisms. Changes in cellular membrane surface potential have been linked with changes in cellular metabolism and motility. Therefore, KPFM represents a powerful tool that can be utilized to examine the changes of microbial membrane surface potential upon adhesion to various substrate surfaces. In this study, we demonstrate the procedure to characterize the surface potential of individual methicillin-resistant Staphylococcus aureus USA100 cells on stainless steel and gold using KPFM. Video LinkThe video component of this article can be found at

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Changes in Plasma Membrane Surface Potential of PC12 Cells as Measured by Kelvin Probe Force Microscopy

Cited by 26 publications

References 35 publications

Direct investigation of current transport in cells by conductive atomic force microscopy

Direct investigation of current transport in cells by conductive atomic force microscopy

A hemicellulose‐bound form of silicon inhibits cadmium ion uptake in rice (Oryza sativa) cells

Surface Potential Measurement of Bacteria Using Kelvin Probe Force Microscopy

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