By Michiel Mueller
This ebook offers a finished account of the idea of picture formation in a confocal fluorescence microscope in addition to a realistic instruction to the operation of the tool, its barriers, and the translation of confocal microscopy facts. The appendices supply a short connection with optical idea, microscopy-related formulation and definitions, and Fourier theory.
- Symbols and abbreviations
- Confocal fluorescence microscopy
- sensible limits
- Miscellaneous topics
- Appendix A: parts of optical theory
- Appendix B: formulation, family and definitions
- Appendix C: Fourier theory
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Extra info for Introduction to Confocal Fluorescence Microscopy, Second Edition
To illustrate this point, consider the results of a numerical simulation shown in Fig. 13. 13 The influence of Poisson noise between two distributions, shifted spatially such that the maximum of the one coincides with the first minimum of the other (Rayleigh criterion). In this example, on average (a) 10, (b) 100, and (c) 1000 photons are detected from each intensity maximum. The reproducibility of each measurement is illustrated by the three subsequent measurements shown horizontally for each case.
For instance, consider the detection of fluorescence emitted from a sample. 12 The Rayleigh criterion. Two components of equal intensity are resolved when the maximum of one coincides with the first minimum of the other. This results in an effective drop of the intensity between the two maxima of ∼20%. Confocal Fluorescence Microscopy 15 domly distributed over time. To determine the average fluorescence rate, the total number of photons emitted over a given time interval can be measured. Because of the random arrival of photons at the detector, repeating the measurement will yield a distribution of values.
17 mm. For some air- and water-immersion objectives, a special correction collar permits correction of spherical aberration for various cover-glass thicknesses. Note that additional spherical aberration is introduced when the refractive index of the object does not match that of the immersion medium (see Sec. 4). 4 Microscope objective types Microscope manufacturers have introduced various types of microscope objectives that compensate for various orders of optical aberration. In addition to spherical aberration, chromatic aberration and field curvature are the most important aberrations.
Introduction to Confocal Fluorescence Microscopy, Second Edition by Michiel Mueller