Genetic fluorescent labeling

Immunofluorescence and Genetic Fluorescent Labeling Techniques in the Dmsophila Nervous System... 

Immunofluorescence (IF) and genetic fluorescent labeling have become standard techniques to study the anatomy, fimction, and development of the Drosophila nervous system. This chapter provides an introduction into these techniques and is aimed to the novice in the field. Besides standard protocols for staining in whole mounts and vibratome sections, we give background information on usefiil antibodies and fly lines and provide guidelines on how to present IF data. We also introduce into the use of neuronal landmarks as a tool for precise and detailed anatomical descriptions. 

The STR data generated at NIST were based on amplifying i.o ng of the genomic DNA with fluorescent labelled primers. The PCR amplified products were analyzed by slab gel electrophoresis followed by imaging with a Molecular Dynamics Fluorl-mager 595 or by capUlary electrophoresis using a PE-ABI 310 Genetic Analyzer. 

There are literally hundreds of markers that are currently available for the mouse and human than can be used to characterize lymphoid and myeloid cells and subsets in primary and secondary lymphoid organs. Many of the markers expressed in mammals are highly conserved across species and have been designated as genetic clusters of differentiation (CD). CDs can be identified with fluorescently labeled monoclonal antibodies. As presented previously, when combined with other fluorescent probes, important information on intracellular biochemistry and cell function can be obtained. Many of the biochemical markers used by immunotoxicologists are common to other... 

TPA (t-PA) tissue-type plasmogen activator, tracer A substance that once introduced into the body can be followed, e.g. due to radio or fluorescent labelling, transaminase (aminotransferase) An enzyme that catalyses the transfer of an amino group for an amino acid to a keto acid to form another amino acid, transcript The RNA that is synthesized by RNA polymerase on a DNA or RNA template, transcriptase An enzyme that catalyses transcription, transcription The mechanism by which information contained in the genetic code is transferred from DNA to RNA.i.e. is transcribed. 

All sequencing instrument represent complex systems that requires considerable manual handling by the operator [ 14,15], Its use requires considerable experience both of the principles of operation that the various operational phases. ABI 3130 Genetic Analyzer (Applied Biosystems) is a four capillary electrophoresis system that uses fluorescently labeled dyes for detection of DNA (http //products.invitrogen.com/ivgn/product/313001). 

A EXPERIMENTAL FIGURE 10-26 During interphase human chromosomes remain in specific domains in the nucleus. Fixed interphase human lymphocytes were hybridized in situ to biotin-labeled probes specific for sequences along the full length of human chromosome 7 and visualized with fluorescently labeled avidin. In the diploid cell shown here, each of the two chromosome 7s is restricted to a territory or domain within the nucleus, rather than stretching throughout the entire nucleus. [From R Lichter et al., 1988, Hum. Genet. 80 224.]... 

Fluorescent spectroscopy, because of its high level of sensitivity, has long been a powerful method for studying protein behavior. Site-specific attachment of fluorophores to a unique cysteine in a protein of interest is a traditional route for the production of fluorescent proteins. In addition, the discovery of fluorescent proteins, such as the green fluorescent protein (GFP) from the jellyfish Aequorea victoria [62], has provided a genetic approach for the production of fluorescently labeled proteins. Both these methods, however,... 

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