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PubMed · 3325697

Retinal wholemounts: a simple method for precise mapping.

P P Giorgi·M L Graydon
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Abstract

A simple and inexpensive method to analyse retinal wholemounts is described. It allows the definition of regularly spaced sites to map regional specializations of the retina under light microscopy. After attaching a piece of translucent millimetre graph paper behind the slide holding the preparation, the two grid patterns of the graph paper and of the eye-piece grid are used in combination for the definition of sites for analysis and the return to specific areas as small as 50-100 microns2. The present method is simpler and offers several advantages when compared to others based on X-Y movements of the microscope stage or on photographic montage.

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Evaluation Studies as TopicHistological TechniquesRetinaRetinal Ganglion CellsJournal ArticleResearch Support, Non-U.S. Gov't

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BibTeXRIS

P P Giorgi, M L Graydon. 1987. Retinal wholemounts: a simple method for precise mapping.. https://doi.org/10.1016/0165-0270(87)90005-7

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Characterization of visible dyes for four-decay fluorescence detection in DNA sequencing.

Dyes of several classes were investigated as candidates for use in a multiplex, four-decay fluorescence detection scheme for DNA sequencing. The dyes include nitrobenzofuran dyes, rhodamine dyes, fluorescein dyes, cyanine dyes, Nile Red, and BODIPY dyes. Based on the results of fluorescence spectral and lifetime studies, an initial set of four dyes was selected for further study: NBD-aminohexanoic acid (NBD-HA, r = 1.1 ns), tetramethyl-rhodamine, methyl ester (r = 2.2 ns), rhodamine green (r = 4.3 ns), and BODIPY 505/515 (r = 5.9 ns). Limits of lifetime detection of the four dyes were investigated, and lifetime resolution was demonstrated for mixtures of the free dyes in batch solution. Lifetime of dye-labeled DNA primers also were determined in batch solution and detected on-the-fly in capillary electrophoresis (CE). Conjugation of the dyes to DNA improved the resolution of their individual lifetimes in mixtures in batch measurements. When attached to the primer, tetramethyl-rhodamine exhibited biexponential decay with a dominant lifetime of 3.8 ns, making it unsuitable for four-decay sequencing. Contact with the CE gel lengthened the lifetime of NBD-HA-labeled primer from 1.3 to 2.1 ns but did not affect the lifetimes of the other dyes. Lifetime detectability of labeled primers at individual points along an electrophoretic peak in the attomole range.

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