影像系统

Many cells undergo directed cell migration in response to external cues in a process known as chemotaxis. This ability is essential for many single-celled organisms to hunt and mate, the development of multicellular organisms, and the functioning of the immune system. Because of their rela ...

Three-dimensional (3D) cell cultures are important tools in cell biology research and tissue engineering because they more closely resemble the architectural microenvironment of natural tissue, compared to standard two-dimensional cultures. Microscopy techniques that ...

During the past 10 years, major developments in live-cell imaging methods have accompanied growing interest in the application of microfluidic techniques to biological imaging. The broad design possibilities of microfabrication and its relative ease of implementation have led ...

Oxygen is essential for survival of aerobic organisms. Sensing changes in the environmental oxygen concentration and appropriate adaptation to such changes are essential for organisms to survive. Hypoxia inducible factor 1 (HIF-1) is the key transcription factor in controlling the ...

Many parameters reflecting mitochondrial function and metabolic status of the cell, including the mitochondrial membrane potential, reactive oxygen species, ATP, NADH, ion gradients, and ion fluxes (Ca2+, H+), are amenable for analysis by live cell imaging and are widely used in many labs. H ...

Cell surface receptors are crucial in the regulation of a wide variety of signalling responses to extracellular stimuli such as soluble growth factors or matrix proteins. To respond effectively to rapidly changing environmental cues, many receptors are rapidly endo- or exo-cytosed to e ...

Detailed practical information is provided with emphasis on mapping cytosolic and mitochondrial pH, mitochondrial Na+, and briefly also aspects related to mitochondrial Ca2+ measurements in living cells, as grown on (un)coated glass coverslips. This chapter lists (laser scanni ...

Protein functions rely on their ability to engage in specific protein–protein interactions and form complexes that are dynamically regulated by stimuli. Bioluminescence resonance energy transfer (BRET) is a highly sensitive technique, which allows monitoring of interaction ...

The changes that occur in electrochemical gradients across biological membranes provide us with invaluable information on physiological responses, pathophysiological processes and drug actions/toxicity. This chapter aims to provide researchers with sufficient inf ...

Many fluorescent probes depend on the fluorescence resonance energy transfer (FRET) between fluorescent protein pairs. The efficiency of energy transfer becomes altered by conformational changes of a fused sensory protein in response to a cellular event. A structure-based appro ...

Fluorescent imaging techniques are powerful tools that aid in studying protein dynamics and membrane domains and allow for the visualization and data collection of such structures as caveolae and clathrin-coated pits, key players in the regulation of cell communication and signali ...

Peroxynitrite (ONOO–) and hypochlorous acid (HOCl) are two highly reactive oxygen species generated in biological systems. The overproduction of peroxynitrite or hypochlorous acid is implicated in a broad array of human pathologies including vascular, immunological, and neur ...

Photo-activatable (caged) probes are powerful research tools for biological investigation. The superb maneuverability of a light beam allows researchers to activate caged probes with pinpoint accuracy. Recent developments in caging chemistry and two-photon excitation te ...

To follow the cell division cycle in the living state, certain biological activity or morphological changes must be monitored keeping the cells intact. Mitotic events from prophase to telophase are well defined by morphology or movement of chromatin, nuclear envelope, centrosomes, and ...

Phagocytosis is the process whereby specialized leukocytes ingest large particles. This is an extremely dynamic and localized process that requires the recruitment to the sites of ingestion of numerous effector proteins, together with extensive lipid remodelling. To investig ...

Fluorescence imaging is an important tool for molecular biology research. There is a wide array of fluorescent labels and activatable probes available for investigation of biochemical processes at a molecular level in living cells. Given the large number of potential imaging agents and ...

For more than a decade, fluorescence resonance energy transfer (FRET) imaging methods have been developed to study dynamic interactions between molecules at the nanometer scale in live cells. Here, we describe a protocol to measure FRET by the acceptor-sensitized emission method as det ...

The analysis of live cells using automated fluorescence microscopy systems on an industrial scale is known as high content screening/analysis (HCS/A). Its development has been driven both by the demands of compound screening in the drug discovery industry and by the promise of whole genome f ...

Since the discovery of the diffraction barrier in the late nineteenth century, it has been commonly accepted that with far-field optical microscopy it is not possible to resolve structural details considerably finer than half the wavelength of light. The emergence of STED microscopy sho ...

Inositol trisphosphate and cyclic ADP-ribose, main intracellular Ca2+ messengers, induce release from the intracellular Ca2+ stores via inositol trisphosphate and ryanodine receptors, respectively. Recently, studies using novel messenger nicotinic acid adenine dinuc ...