These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer‐reviewed by leading experts in the field, making this an essential research companion.
14 November 2007 in Analytical and Bioanalytical Chemistry
The last decade has witnessed the development of a variety of metal nanoparticle-based techniques for DNA detection. High sensitivity and specificity, miniaturization, and cost-efficient detection are problems addressed by the use of nanoparticle labels in heterogeneous DNA detection schemes. The small label size, established bioconjugation chemistry, and the unusual optical and electrical properties of metal nanoparticles make them unique tools for DNA detection. This paper reviews the different physical characteristics of metal nanoparticles and their implementation in assays. It covers various optical as well as gravimetric, electrochemical and electrical methods for analysing nanoparticle-labelled analytes, and particularly DNA, at sensing surfaces.
Near-infrared laser pulses of a compact 80-MHz femtosecond laser source at 800 nm, a mean power of 15–100 mW, 170-fs pulse width, and millisecond beam dwell times at the target have been used for multiphoton-mediated nanoprocessing of human chromosomes. By focusing of the laser beam with high-numerical-aperture objectives of a scanning microscope to diffraction-limited spots and with light intensities of terawatts per cubic centimeter, precise submicrometer holes and cuts in human chromosomes have been processed by single-point exposure and line scans. A minimum FWHM cut size of during a partial dissection of chromosome 1, which is below the diffraction-limited spot size, and a minimum material removal of were determined by a scanning-force microscope. The plasma-induced ablated material corresponds to of the chromosome 1 volume and to base pairs of chromosomal DNA. A complete dissection could be performed with FWHM cut sizes below 200 nm. High-repetition-frequency femtosecond lasers at low mean power in combination with high-numerical-aperture focusing optics appear therefore as appropriate noncontact tools for nanoprocessing of bulk and (or) surfaces of transparent materials such as chromosomes. In particular, the noninvasive inactivation of certain genomic regions on single chromosomes within living cells becomes possible.