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Observation of Oxidative Stress on Yeast Cells

Before the advent of the atomic force microscope (AFM), scanning electron microscopy (SEM) was used to obtain high-resolution visualizations of the surface of biological samples. Normally, to scan samples of yeast cells, each preparation was coated with a film of evaporated gold approx 20 nm ...

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Lymphoblastoid Cells Exposed to Low-Frequency Magnetic Fields: Study by Atomic Force Microscopy

Since the appearance of the first pioneering article in the 1970s, particular efforts have been made to study the effect of exposure to electric and magnetic fields (EMFs) on living matter (1–10). The initial interest in radio frequency and microwaves has shifted to include the nonthermal, and es ...

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Atomic Force Microscopy of -Amyloid: Static and Dynamic Studies of Nanostructure and Its Formation

Ordered aggregation of the β-amyloid (Aβ) peptide in the brain as plaques consisting of fibrils is an important characteristic of Alzheimer’s disease (AD), a late onset neurodegenerative disease (1). Aβ derives from the endoproteolysis of the amyloid precursor protein (APP), which is a tra ...

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Sample Preparation Method for Observing RNA Polymerase Activity by Atomic Force Microscopy

Transcription is a fundamental biochemical process in which an RNA molecule is synthesized according to its corresponding DNA template.

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How to Build Up Biosensors With the Cantilever of the Atomic Force Microscope

With the advent of the atomic force microscopy (AFM), the study of biological samples has become more realistic because, in most cases, samples are not covered or fixed and this makes it possible to observe them while alive (1,2). This advantage of the AFM prompted a new invention: nanobiosensors using ...

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Measurement of Single Molecular Interactions by Dynamic Force Microscopy

Unbinding forces of weak, noncovalent bonds have been measured by scanning force microscopy (1) or biomembrane force probes (2). Initially, these scanning force microscopy measurements focused on feasibility studies to measure single biomolecular interactions (3–5). Recentl ...

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Laser Capture Microdissection

Laser capture microdissection (LCM) is a technique for isolating pure cell populations from a heterogeneous tissue section or cytological preparation via direct visualization of the cells. This technique is applicable to molecular profiling of diseased and disease-free tissue, ...

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Analysis of Asbestos-Induced Gene Expression Changes in Bronchiolar Epithelial Cells Using Laser Capture Microdissection and Quantitative Reverse Tran

Laser capture microdissection (LCM) enables the removal of discrete microstructures or cell types from properly prepared histological sections. Extraction of RNA from microdissected tissue followed by quantitative reverse transcriptase-polymerase chain (QRT-PCR) r ...

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New Approaches to Fluorescence In Situ Hybridization

Fluorescence in situ hybridization (FISH) is a nonisotopic labeling and detection method that provides a direct way to determine the relative location or copy number of specific DNA sequences in nuclei or chromosomes. With recent advancements, this technique has found increased appl ...

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Microarray Image Scanning

Of the technologies available for measuring gene expression, microarrays using cDNA targets is one of the most common and well-developed high-throughput techniques. With this technique, the expression levels of thousands of genes are measured simultaneously. DNA probes are immob ...

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Near-Field Scanning Optical Microscopy in Cell Biology and Cytogenetics

Light microscopy has proven to be one of the most versatile analytical tools in cell biology and cytogenetics. The growing spectrum of scientific knowledge demands a continuous improvement of the optical resolution of the instruments. In far-field light microscopy, the attainable res ...

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Porosome: The Fusion Pore Revealed by Multiple Imaging Modalities

Secretion occurs in all cells of multicellular organisms and involves the delivery of secretory products packaged in membrane-bound vesicles to the cell exterior. Specialized cells for neuro-transmission, enzyme secretion, or hormone release utilize a highly regulated secre ...

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Secretory Vesicle Swelling by Atomic Force Microscopy

The swelling of secretory vesicles has been implicated in exocytosis, but the underlying mechanism of vesicle swelling remained unknown. Earlier studies from our laboratory demonstrated the association of the α-subunit of heterotrimeric GTP-binding protein G�i3 with zymogen g ...

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Imaging and Probing Cell Mechanical Properties With the Atomic Force Microscope

This chapter describes the use of the atomic force microscope (AFM) to probe and map out regional variations in apparent elastic properties of living cells. The importance of mechanics in the field of cell biology is becoming more widely appreciated, and the AFM has unique advantages for cell mec ...

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Three-Dimensional Analysis of Single Particles by Electron Microscopy: Sample Preparation and Data Acquisition

Electron microscopy of single particles has recently become a very popular field in both biological and material sciences. It might be difficult for a novice researcher new to this field to know how to start tackling a new project. This chapter is designed to serve as a guideline for anyone starting a new ...

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Reflection Contrast Microscopy: The Bridge Between Light and Electron Microscopy

Reflection contrast microscopy (RCM) is a light microscopic method to image cells at high definition and enhanced sensitivity compared to conventional bright-field microscopy. RCM images have very high contrast, which makes them easily applicable for digital image analysis. Bec ...

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Molecular Beacons: Fluorescent Probes for Detection of Endogenous mRNAs in Living Cells

A novel approach for detecting nucleic acid in solution has been adopted for real-time imaging of native mRNAs in living cells. This method utilizes hybridization probes, called “molecular beacons,” that generate fluorescent signals only when they are hybridized to a complementary ta ...

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Three-Dimensional Reconstruction of Single Particles in Electron Microscopy: Image Processing

Three-dimensional electron microscopy of single macromolecular assemblies has made large strides forward over the last decade. A large number of image processing techniques have been developed and many have found general distribution. For the proper usage of the wide range of availa ...

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Second-Harmonic Imaging of Collagen

Molecules that have no center of symmetry are able to convert light to its second harmonic, at twice the frequency and half the wavelength. This only happens with any efficiency at very high light intensities such as are given by a pulsed laser, and because the efficiency of the process depends on the squa ...

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A New Microbiopsy System Enables Rapid Preparation of Tissue for High-Pressure Freezing

A microbiopsy system was developed to overcome long sampling times for tissues before they are cryo-fixed by high-pressure freezing. A commercially available biopsy gun was adapted to the needs of small-organ excisions, and biopsy needles were modified to allow small samples (0.6 mm k 1.2 mm � 0.3 m ...

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