Imaging a specific mRNA in pollen with atomic force microscopy

Kim, Jung Sook; Park, Yu Shin; Nam, Hong Gil; Park, Chong-Yun

doi:10.1039/c5ra00199d

Cited by 5 publications

(5 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Additionally, sectioning of fixed cells and tissues would enable the inner part of cell and specific cellular compartments to be examined. 55 We believe that this new analytical approach opens a new avenue for understanding the biological role of miRNAs and their cell-to-cell variation. In particular, the detection of small amounts of miRNA biomarkers in biological samples would enable this approach to serve as a cancer diagnostic tool.…”

Section: ■ Discussionmentioning

confidence: 99%

“…Furthermore, with high-speed AFM machines, ,,− it is expected that the whole body of a single cell can be visualized efficiently. Additionally, sectioning of fixed cells and tissues would enable the inner part of cell and specific cellular compartments to be examined . We believe that this new analytical approach opens a new avenue for understanding the biological role of miRNAs and their cell-to-cell variation.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Visualization and Quantification of MicroRNA in a Single Cell Using Atomic Force Microscopy

et al. 2016

Self Cite

View full text Add to dashboard Cite

MicroRNAs (miRNAs) play critical roles in controlling various cellular processes, and the expression levels of individual miRNAs can be considerably altered in pathological conditions such as cancer. Accurate quantification of miRNA at the single-cell level will lead to a better understanding of miRNA function. Here, we present a direct and sensitive method for miRNA detection using atomic force microscopy (AFM). A hybrid binding domain (HBD)-tethered tip enabled mature miRNAs, but not premature miRNAs, to be located individually on an adhesion force map. By scanning several sections of a micrometer-sized DNA spot, we were able to quantify the copy number of miR-134 in a single neuron and demonstrate that the expression was increased upon cell activation. Moreover, we visualized individual miR-134s on fixed neurons after membrane removal and observed 2-4 miR-134s in the area of 1.0 × 1.0 μm(2) of soma. The number increased to 8-14 in stimulated neurons, and this change matches the ensemble-averaged increase in copy number. These findings indicate that miRNAs can be reliably quantified at the single cell level with AFM and that their distribution can be mapped at nanometric lateral resolution without modification or amplification. Furthermore, the analysis of miRNAs, mRNAs, and proteins in the same sample or region by scanning sequentially with different AFM tips would let us accurately understand the post-transcriptional regulation of biological processes.

show abstract

Section: ■ Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Visualization and Quantification of MicroRNA in a Single Cell Using Atomic Force Microscopy

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…For example, intermediate filaments offer mechanical stability to cells when exposed to mechanical stress, and act as a support when the other cytoskeletal filaments cannot keep the structural integrity of the cells [131]. AFM can also be applied for the mapping of target RNA distribution using probe DNA-immobilized AFM tips [132], which is fundamentally important to understand the regulatory mechanisms underlying cellular and tissue differentiation [133].…”

Section: Applicationsmentioning

confidence: 99%

Nanoindentation of Soft Biological Materials

2018

View full text Add to dashboard Cite

Nanoindentation techniques, with high spatial resolution and force sensitivity, have recently been moved into the center of the spotlight for measuring the mechanical properties of biomaterials, especially bridging the scales from the molecular via the cellular and tissue all the way to the organ level, whereas characterizing soft biomaterials, especially down to biomolecules, is fraught with more pitfalls compared with the hard biomaterials. In this review we detail the constitutive behavior of soft biomaterials under nanoindentation (including AFM) and present the characteristics of experimental aspects in detail, such as the adaption of instrumentation and indentation response of soft biomaterials. We further show some applications, and discuss the challenges and perspectives related to nanoindentation of soft biomaterials, a technique that can pinpoint the mechanical properties of soft biomaterials for the scale-span is far-reaching for understanding biomechanics and mechanobiology.

show abstract

“…AFM has been applied for the mapping of target RNA distribution using probe DNA-immobilized AFM tips [56,57]. It is of fundamental importance to detect the cellular and tissue distributions of RNA species to understand the regulatory mechanisms underlying cellular and tissue differentiation.…”

Section: Detection and Quantification Of Target Rna Via Afm Force Pro...mentioning

confidence: 99%

“…A similar concept was applied from the same research group to image a specific mRNA in pollen, determining the intracellular localization of the ammonium transporter (AtAMT1;4) mRNA in the sperm cells of Arabidopsis pollen [57]. The AFM probe was modified with cone-shaped dendron molecules, and an oligonucleotide complementary to the mRNA was attached to the apex.…”

Section: Detection and Quantification Of Target Rna Via Afm Force Pro...mentioning

confidence: 99%

Atomic force microscopy of RNA: State of the art and recent advancements

Schön

2018

Seminars in Cell & Developmental Biology

View full text Add to dashboard Cite

Imaging a specific mRNA in pollen with atomic force microscopy

Abstract: Distribution of ammonium transporter mRNA in a sectioned pollen is studied at the higher resolution, and localization of the mRNA in the nucleus of the sperm cells is observed.

Cited by 5 publications

References 36 publications

Visualization and Quantification of MicroRNA in a Single Cell Using Atomic Force Microscopy

Visualization and Quantification of MicroRNA in a Single Cell Using Atomic Force Microscopy

Nanoindentation of Soft Biological Materials

Atomic force microscopy of RNA: State of the art and recent advancements

Contact Info

Product

Resources

About