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Cellular imaging techniques for neuroscience and beyond /
The imaging of small cellular components requires powerful instruments, and an entire family of equipment and techniques based on the confocal principle has been developed over the past 30 years. Such methods are commonly used by neuroscience researchers, but the majority of these users do not have...
| Clasificación: | Libro Electrónico |
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| Otros Autores: | |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
Amsterdam :
Elsevier/Academic Press,
2012.
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| Temas: | |
| Acceso en línea: | Texto completo |
Tabla de Contenidos:
- Front Cover; Cellular Imaging Techniques for Neuroscience and Beyond; Copyright Page; Contents; List of Contributors; 1 Confocal Laser Scanning: of Instrument, Computer Processing, and Men; Introduction; Pinhole, Depth of Focus, and Laser Illumination; When/Why Does One Need a CLSM?; Abbe, Shannon, and Nyquist; Imaging of a 2D Line and Deblurring; Axial Resolution; Resolution and Sampling; Signal Separation, Orders of Magnitude, and Resolution Limits; Confocal Microscopy Further Considered; Cross Talk Awareness; Excitation Cross Talk; Elimination of Cross Talk.
- Biological Objects Translated to PixelsHigh-probability Determination of Diameter; Best-Fit Object 3D Recognition; Automated Objective Threshold Analysis; Why Does a 3D Reconstructed Cell Resemble a Pancake?; Touch; Actual Experiment; Computer Software to Define a Contact; Synaptic Contacts: Extra Marker; Colocalization; Conclusion; Acknowledgments; References; 2 Beyond Abbe's Resolution Barrier: STED Microscopy; Introduction; A New Wave of Imaging; STED Microscopy: The Basic Concept; Implementation of STED Microscopy; The Microscope Base; Laser Sources; The Doughnut; Synchronization.
- Objective LensesEmission Detection; Scanning Schemes; Software; Stability Considerations; Checking the Resolution; Sine Qua Non: Speed, Color, Depth, Live Imaging; Temporal Resolution and Imaging Speed; Labeling Strategies; Multicolor Imaging; Depth Penetration and Spatial Resolution; Spatial Resolution in z; Live-cell Imaging; Summary and Outlook; Reversible Saturable Optical Fluorescence Transitions: A More General Principle for Nanoscopy; Other Areas of Development; References; 3 Enhancement of Optical Resolution by 4pi Single and Multiphoton Confocal Fluorescence Microscopy; Introduction.
- The 4pi Principle and SetupMicroscope Alignment; Initial Alignment of the Excitation by Eye; Optimizing the Coverslip Correction Ring; Alignment of the Bottom Lens (XYZ); Alignment of the Second Mirror (XYZ); 4pi Imaging; 4pi Deconvolution; Sample Preparation; Fixation; Selection of Fluorescent Dyes; Microtubule and Microtubule Plus End Imaging; Visualization of DNA; 4pi Imaging of Muntjac Chromosomes; Single-photon Excitation (Measurement of the Redox State in Dopamine Neurons); SYCP3 Axis as a Marker for Chromatin Organization in Mouse Spermatocytes; Microbubbles with Medicine.
- Future of 4pi ImagingAcknowledgment; References; 4 Nano Resolution Optical Imaging Through Localization Microscopy; Introduction; Superresolution Microscopy Techniques; The Concept Behind Localization Microscopy; Matters of Concern; Localization Precision; Linkage Error; Specificity; Labeled Fraction; Labeling Density; The Main Approaches to Single-molecule Localization-based Superresolution Microscopy; Using Fluorescent Proteins: PALM and FPALM; Using a Pair of Interacting Cyanine Dyes: STORM; Using Blinking Dyes: dSTORM; Using High-intensity Light: GSDIM; Using Targeted Molecules: BALM.