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Titolo:
High-definition mapping of neural activity using voltage-sensitive dyes
Autore:
Cinelli, AR;
Indirizzi:
SUNY Hlth Sci Ctr, Dept Anat & Cell Biol, Brooklyn, NY 11203 USA SUNY HlthSci Ctr Brooklyn NY USA 11203 Cell Biol, Brooklyn, NY 11203 USA SUNY Hlth Sci Ctr, Video Imaging Facil, Brooklyn, NY 11203 USA SUNY Hlth Sci Ctr Brooklyn NY USA 11203 ing Facil, Brooklyn, NY 11203 USA
Titolo Testata:
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY
fascicolo: 4, volume: 21, anno: 2000,
pagine: 349 - 372
SICI:
1046-2023(200008)21:4<349:HMONAU>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
SALAMANDER OLFACTORY-BULB; CENTRAL-NERVOUS-SYSTEM; MONKEY STRIATE CORTEX; NEURONAL-ACTIVITY; FLUORESCENCE MICROSCOPY; VIDEO-RATE; TEMPORAL PROPERTIES; OPTICAL RECORDINGS; LIGHT-MICROSCOPE; 3 DIMENSIONS;
Keywords:
video microscopy; fluorescence microscopy; optical recording; voltage-sensitive dye; deconvolution; digital confocal microscopy;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
53
Recensione:
Indirizzi per estratti:
Indirizzo: Cinelli, AR SUNY Hlth Sci Ctr, Dept Anat & Cell Biol, Brooklyn, NY 11203 USA SUNY Hlth Sci Ctr Brooklyn NY USA 11203 rooklyn, NY 11203 USA
Citazione:
A.R. Cinelli, "High-definition mapping of neural activity using voltage-sensitive dyes", METHODS, 21(4), 2000, pp. 349-372

Abstract

The distribution of patterns of activity in different brain structures hasbeen related to the encoding and processing of sensory information. Consequently, it is important to be able to image the distribution of these patterns to understand basic brain functions. The spatial resolution of voltage-sensitive dye (VSD) methods has recently been enhanced considerably by the use of video imaging techniques. The main factor that now hampers the resolution of VSD patterns is the inherent limitation of the optical systems. Unfortunately, the intrinsic characteristics of VSD images impose important limitations that restrict the use of general deconvolution techniques. To overcomes this problem, in this study an image restoration procedure has beenimplemented that takes into consideration the limiting characteristics of VSD signals. This technique is based on applying a set of imaging processing steps. First, the signal-to-noise (S/N) ratio of the images was improved to avoid an increase in the noise levels during the deconvolution procedures. For this purpose, a new filter technique was implemented that yielded better results than other methods currently used in optical imaging. Second, focal plane images were deconvolved using a modification of the well-known nearest-neighbor deconvolution algorithm. But to reduce the light exposure of the preparation and simplify image acquisition procedures, adjacent image planes were modeled according to the in-focus image planes and the empirical point spread function (PSF) profiles. Third, resulting focal plane responses were processed to reduce the contribution of optical responses that originate in distant image planes. This method was found to be satisfactory under simulated and real experimental conditions. By comparing the restoredand unprocessed images, it was clearly demonstrated that this method can effectively remove the out-of-focus artifacts and produce focal plane imagesof better quality. Evaluations of the tissue optical properties allowed assessment of the maximum practical optical section thickness using this deconvolution technique in the optical system tested. Determination of the three-dimensional PSF permitted the correct application of deconvolution algorithms and the removal of the contaminating light arising from adjacent as well as distant optical planes. The implementation of this deconvolution approach in salamander olfactory bulb allowed the detailed study of the laminardistribution of voltage-sensitive changes across the bulb layer. It is concluded that (1) this deconvolution procedure is well suited to deconvolved low-contrast images and offers important advantages over other alternatives; (2) this method can be properly used only when the tissue optical properties are first determined; (3) high levels of light scattering in the tissuereduce the optical section capabilities of this technique as well as otherdeconvolution procedures; and (4) use of the highest numerical aperture inthe objectives is advisable because this improves not only the light-collecting efficiency to detect poor-contrast images, but also the spatial frequency differences between adjacent image planes. Under this condition it is possible to overcome some of the limitations imposed by the light scattering/birefringence of the tissue. (C) 2000 Academic Press.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 31/03/20 alle ore 08:29:52