Abstract:
:We describe a novel procedure for combining immunocytochemistry with in situ hybridisation. In contrast to previously published procedures, the technique involves immunofluorescence followed by in situ hybridization and is particularly suitable for antigens which are labile or sensitive to in situ hybridization processing. We have evaluated the technique using 5-hydroxytryptamine (5-HT, serotonin) immunofluorescence and neuropeptide in situ hybridization employing 35S-labelled oligonucleotide probes. Successful double labelling was obtained and showed that galanin messenger RNA (mRNA) is expressed by 5-HT immunoreactive cells in the dorsal raphe nucleus of the rat. In contrast, somatostatin mRNA in the same region is expressed by a separate non-serotonergic cell population. Double-labelled preparations produced using this technique can be conveniently viewed using epipolarised combined with epifluorescent illumination. Careful analysis of procedural variables revealed that it is not possible to carry out high-sensitivity 5-HT immunocytochemistry following in situ hybridization. The immunostaining is much poorer on slide-mounted sections than on free-floating sections, and 5-HT appears to be lost during the in situ hybridization steps of dehydration/delipidation and incubation in hybridization buffer. The procedure we describe avoids these problems but with a slight loss of in situ hybridization sensitivity.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Priestley JV,Wotherspoon G,Savery D,Averill S,Rattray Mdoi
10.1016/s0165-0270(05)80011-1subject
Has Abstractpub_date
1993-06-01 00:00:00pages
99-110issue
1-2eissn
0165-0270issn
1872-678Xpii
S0165-0270(05)80011-1journal_volume
48pub_type
杂志文章abstract:BACKGROUND:Human neurons (hNT neurons), obtained from the NTera2/D1 precursor cell line, are highly valued by many neuroscientists as isolation of adult human primary neuronal cells continues to elude us. hNT neurons are generated by differentiation of the NT2 precursors for a period of 4 weeks followed by 2 weeks of m...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2014.02.004
更新日期:2014-04-30 00:00:00
abstract::Vertex analysis defines dichotomous networks in terms of the relative frequencies of pendant (Vp) and nodal (Vd) vertices. Va, Vb and Vc comprise the 3 classes of Vd which connect 2, 1 and 0 Vp respectively. The vertex ratio (VR) of Va/Vb may be used to define topology since Va and Vc approach equality in large networ...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(86)90118-4
更新日期:1986-10-01 00:00:00
abstract::The cytochemical localization of acetylcholinesterase activity at motor endplates is commonly employed prior to electron microscopic examination. Methods which involve the use of thiocholine esters often lead to a loss of resolution due to the spread to surrounding tissues of an extremely dense reaction product. A met...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(82)90031-0
更新日期:1982-11-01 00:00:00
abstract:BACKGROUND:The local field potential (LFP) is usually calculated from current sources arising from transmembrane currents, in particular in asymmetric cellular morphologies such as pyramidal neurons. NEW METHOD:Here, we adopt a different point of view and relate the spiking of neurons to the LFP through efferent synap...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2020.108871
更新日期:2020-10-01 00:00:00
abstract::Electrophysiological recording provides important evidence for positive identification of many neurons in gastropods. We describe a technique which combines intracellular recording and injection of a persistent, non-fluorescent dye (Fast Green) with subsequent histofluorescence treatment using a modification of the wh...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(85)90036-6
更新日期:1985-08-01 00:00:00
abstract::The amplitude of the maximal direct motor response (Mmax) elicited by supramaximal peripheral nerve stimulation can vary with time and with changes in muscle length. We sought to investigate the variability in the amplitude of Mmax in the human wrist flexors (flexor carpi radialis, FCR) at a constant joint position du...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2005.03.016
更新日期:2005-12-15 00:00:00
abstract::Some basic methodological issues concerning the use of the fluorescent tracers Fast blue (FB) and Diamidino yellow (DY) were studied using the projections of the red nucleus to the nucleus interpositus anterior (NIA) of the cerebellum of the cat. On standard Nissl-stained sections, it was possible to delineate 4 disti...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(87)90100-2
更新日期:1987-09-01 00:00:00
abstract::An important characterization of neural spiking is the ratio of the variance to the mean of the spike counts in a set of intervals--the Fano factor. For a Poisson process, the theoretical Fano factor is exactly one. For simulated or experimental neural data, the sample Fano factor is never exactly one, but often appea...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2010.04.012
更新日期:2010-06-30 00:00:00
abstract::Superpositioning of multiple 3-dimensional digital volumes is essential to extending neuroimaging capabilities from single-subject analysis to group evaluation. In this study, 6 rat heads were cryoplaned and digital images of the specimen blockface were organized into data volumes. The midline plane of each brain guid...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(93)90007-e
更新日期:1993-11-01 00:00:00
abstract:BACKGROUND:Neural coding of sound information is often studied through frequency tuning curve (FTC), spectro-temporal receptive field (STRF), post-stimulus time histogram (PSTH), and other methods such as rate functions. These methods, despite providing a robust characterization of auditory responses in their specific ...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2020.108682
更新日期:2020-05-15 00:00:00
abstract::A simple method which allows easy quantification of neuronal and glial sub-populations in primary neuronal tissue culture is described. The method takes advantage of the fact that the staining of neuronal and glia nuclei with Hoechst 33258 is distinctive and characteristic of the cell type. Double-staining experiments...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(94)00207-w
更新日期:1995-07-01 00:00:00
abstract::We investigated the usefulness of YFP-H transgenic mice [Neuron 28 (2000) 41] which express yellow fluorescent protein (YFP) in a restricted subset of neurons to study Wallerian degeneration in the PNS. Quantification of YFP positive axons and myelin basic protein (MBP) immunocytochemistry revealed that YFP was random...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2003.10.016
更新日期:2004-03-15 00:00:00
abstract:BACKGROUND:Image segmentation is often imperfect, particularly in complex image sets such z-stack micrographs of slice cultures and there is a need for sufficient details of parameters used in quantitative image analysis to allow independent repeatability and appraisal. NEW METHOD:For the first time, we have criticall...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2017.12.002
更新日期:2018-02-01 00:00:00
abstract::We developed a high speed voice coil based whisker stimulator that delivers precise deflections of a single whisker or group of whiskers in a repeatable manner. The device is miniature, quiet, and inexpensive to build. Multiple stimulators fit together for independent stimulation of four or more whiskers. The system c...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2010.12.008
更新日期:2011-03-15 00:00:00
abstract::The cost of electronic equipment can be a critical barrier to including neurophysiology exercises in biology teaching programs. We describe the construction of a simple and inexpensive AC preamplifier with performance comparable to that of commercial products. The amplifier consists of two integrated circuits in five ...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/s0165-0270(01)00328-4
更新日期:2001-03-30 00:00:00
abstract:BACKGROUND:In clinical practice, small myelinated sensory fibers, Aδ-fibers, conveying mainly pain and temperature sensations, cannot be examined with available nerve conduction study techniques. Currently, these fibers can only be examined with experimental or very specialized and not commonly available nerve conducti...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2020.109045
更新日期:2020-12-24 00:00:00
abstract::Recent studies highlighted the disagreement between the typical number of neurons observed with extracellular recordings and the ones to be expected based on anatomical and physiological considerations. This disagreement has been mainly attributed to the presence of sparsely firing neurons. However, it is also possibl...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2012.07.010
更新日期:2012-10-15 00:00:00
abstract::A combination of two procedures, embedding of specimens and horseradish peroxidase (HRP) histochemistry on mounted serial sections, is proposed for three-dimensional reconstruction of neuronal projections. To clarify the intrinsic organization of the peripheral nervous system, this simple method is more adequate than ...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(86)90130-5
更新日期:1986-09-01 00:00:00
abstract::This report describes a digitally controlled system for use in auditory physiology in which signal generation and data acquisition are under the control of a single microcomputer. The system is designed to generate the complex waveforms required in neuroethological studies, and is able to generate these signals at fre...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(91)90136-n
更新日期:1991-01-01 00:00:00
abstract:BACKGROUND:Transcranial magnetic stimulation (TMS) can be paired with functional magnetic resonance imaging (fMRI) in concurrent TMS-fMRI experiments. These multimodal experiments enable causal probing of network architecture in the human brain which can complement alternative network mapping approaches. Critically, me...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2019.108400
更新日期:2019-11-01 00:00:00
abstract::We present a numerical method which provides the ability to analyze digitized microscope images of retinal explants and quantify neurite outgrowth. Few parameters are required as input and limited user interaction is necessary to process an entire experiment of images. This eliminates fatigue related errors and user-r...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/s0165-0270(03)00017-7
更新日期:2003-04-15 00:00:00
abstract:BACKGROUND:Intracranial electrical recordings (iEEG) and brain stimulation (iEBS) are invaluable human neuroscience methodologies. However, the value of such data is often unrealized as many laboratories lack tools for localizing electrodes relative to anatomy. To remedy this, we have developed a MATLAB toolbox for int...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2017.01.022
更新日期:2017-04-01 00:00:00
abstract::In this paper we present an efficient method to generate realistic simulations of extracellular recordings. The method uses a hybrid and computationally simple approach, where the features of the background noise arise naturally from its biophysical process of generation. The generated data resemble the characteristic...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2009.08.017
更新日期:2009-11-15 00:00:00
abstract::An inexpensive system for quantitative electroencephalogram (EEG) analysis over long time periods is described. This system continuously analyses EEG for intensity versus frequency content, and saves averaged 'data reduced' intensity spectra to disk at regular intervals. The method had adequate frequency resolution fo...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(90)90066-o
更新日期:1990-04-01 00:00:00
abstract::A method for automatically analyzing cortical signals recorded with voltage-sensitive dyes and a photodiode array is described. First, a forward-backward non-linear filtering technique is used to eliminate the background noise and preserve the fast transients of the signals. Then the filtered signals are deconvoluted ...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(95)00105-0
更新日期:1995-12-01 00:00:00
abstract:BACKGROUND:Local shape complexity can be biologically meaningful as a marker of disease, trauma, or change in brain structure over time. Fractal dimensionality (FD) is currently the dominant measure of local shape complexity used in neuroimaging but its limitations are not well understood. NEW METHOD:Elliptical Fourie...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2019.05.009
更新日期:2019-07-15 00:00:00
abstract::Skeletal muscle is a convenient model system for studying basic questions on the neural regulation of synaptogenesis and of many properties of sarcolemma and contractile apparatus. The study of the neural signals involved in a particular regulation and of the mediating intracellular pathways, requires the chronic appl...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/s0165-0270(00)00352-6
更新日期:2001-04-30 00:00:00
abstract::A method is described for producing electrolytically sharpened and lacquer-insulated steel microelectrodes and for their attachment to a multibarrel micropipet assembly by light-curing bonding compound. This electrode complex is specially designed for recordings from deep brain structures. It permits the analysis of t...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(88)90007-6
更新日期:1988-04-01 00:00:00
abstract::The present study evaluated high-speed chronoamperometry as a method for measuring the clearance of serotonin (5-HT) from extracellular space in vivo. Male Sprague-Dawley rats were anaesthetized and a Nafion-coated, carbon fiber electrode, attached to a multibarrel pipette, was lowered into the subgranular layer of th...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/s0165-0270(97)00144-1
更新日期:1997-12-30 00:00:00
abstract:BACKGROUND:It has previously been reported that EEG sigma (10-15 Hz) activity during sleep exhibits infraslow oscillations (ISO) with a period of 50 s. However, a detailed analysis of the ISO of individually identified sleep spindles is not available. NEW METHOD:We investigated basic properties of ISO during baseline ...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2018.12.002
更新日期:2019-03-15 00:00:00