A rapid cell counting method utilising acridine orange as a novel discriminating marker for both cultured astrocytes and microglia.

Abstract:

:Cell culture analyses of growth, morphology and apoptosis commonly require counting of different cell types stained with antibodies to discriminate between them. Previously, we reported the use of l-Leucine methyl ester (l-LME) to prepare purified cultures of type 1 astrocytes with minimal microglia, and staining by GFAP and CD antibodies, respectively. Here, we demonstrate a novel use of acridine orange (AO) for rapid discrimination between these cell types using fluorescence microscopy. AO accumulates in the lysosomes and also binds strongly to nuclear DNA and cytoplasmic/nucleolar RNA. Microglia may contain abundant lysosomes due to known roles in homeostasis and immune response. AO staining of lysosomes was tested at a range of concentrations, and 2.5 microg/mL was most suitable. In agreement with previous reports, microglia treated with AO showed very intense yellow, orange or red granular cytoplasmic staining of lysosomes. Microglia contain a substantially higher number of lysosomes than astrocytes, which have a variable amount. We measured the microglia population at 5.14+/-0.50% in mixed cultures. Thus, these results show AO is a novel discriminatory marker, as microglia were easily observed and counted in clumps on top of the monolayer of astrocytes, providing a rapid alternative to time-consuming and costly antibody-based assays.

journal_name

J Neurosci Methods

authors

Lovelace MD,Cahill DM

doi

10.1016/j.jneumeth.2007.06.009

subject

Has Abstract

pub_date

2007-09-30 00:00:00

pages

223-9

issue

2

eissn

0165-0270

issn

1872-678X

pii

S0165-0270(07)00277-4

journal_volume

165

pub_type

杂志文章
  • Formalin test in mice, a useful technique for evaluating mild analgesics.

    abstract::A modification of the formalin test appropriate for testing of mice is described. Formalin 1 or 5% was injected into the dorsal surface of a hindpaw, and the time the animal spent licking the paw was recorded. On the basis of the response pattern, two distinct periods of intensive licking activity were identified; an ...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/0165-0270(85)90116-5

    authors: Hunskaar S,Fasmer OB,Hole K

    更新日期:1985-06-01 00:00:00

  • Consistent and reproducible slice selection in rodent brain using a novel stereotaxic device for MRI.

    abstract::Typically small animal radiological images are obtained after placing the animal in the center of the imaging device using beds or platforms, and then adjusting the position after obtaining a scout image. Such a process does not permit the reproducible visualization of the same anatomical plane with repeated examinati...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2004.01.003

    authors: Fricke ST,Vink R,Chiodo C,Cernak I,Ileva L,Faden AI

    更新日期:2004-06-15 00:00:00

  • Frequency-based approach to the study of semantic brain networks connectivity.

    abstract::The interactions among cerebral regions involved in semantic word generations are explored through connectivity analysis based on fMRI data through multivariate autoregressive model (MVAR). Connections among the pars triangularis of the left inferior frontal gyrus (L45), the lateral fusiform girus (LFG) and the left m...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2012.10.005

    authors: Bianchi AM,Marchetta E,Tana MG,Tettamanti M,Rizzo G

    更新日期:2013-01-30 00:00:00

  • Evaluation of neuronal numerical density by Dirichlet tessellation.

    abstract::The technique that we describe aims at evaluating the numerical density of cells in highly heterogeneous regions, e.g., nuclei, layers or columns of neurones. Rather than counting the number of neuronal sections ('profiles') in a reference frame, we evaluated the 'free area' which lies around each profile. The X and Y...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/0165-0270(94)90025-6

    authors: Duyckaerts C,Godefroy G,Hauw JJ

    更新日期:1994-01-01 00:00:00

  • Reliability of the microdialysis pump CMA 107 under hyperbaric conditions.

    abstract:OBJECTIVE:Microdialysis measurements of extracellular substances under hyperbaric conditions were manifold used in several investigations. However, to our knowledge there is no analysis, which verified the applicability of microdialysis pumps under hyperbaric conditions. Thus, a goal of this study was to investigate th...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2007.05.001

    authors: Henze D,Köthe L,Scharf A,Clausen T

    更新日期:2007-08-30 00:00:00

  • SWDreader: a wavelet-based algorithm using spectral phase to characterize spike-wave morphological variation in genetic models of absence epilepsy.

    abstract:BACKGROUND:Spike-wave discharges (SWD) found in neuroelectrical recordings are pathognomonic to absence epilepsy. The characteristic spike-wave morphology of the spike-wave complex (SWC) constituents of SWDs can be mathematically described by a subset of possible spectral power and phase values. Morlet wavelet transfor...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2014.12.016

    authors: Richard CD,Tanenbaum A,Audit B,Arneodo A,Khalil A,Frankel WN

    更新日期:2015-03-15 00:00:00

  • Advanced correlation grid: Analysis and visualisation of functional connectivity among multiple spike trains.

    abstract:BACKGROUND:This study analyses multiple spike trains (MST) data, defines its functional connectivity and subsequently visualises an accurate diagram of connections. This is a challenging problem. For example, it is difficult to distinguish the common input and the direct functional connection of two spike trains. NEW ...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2017.05.016

    authors: Masud MS,Borisyuk R,Stuart L

    更新日期:2017-07-15 00:00:00

  • Generation of a neurodegenerative disease mouse model using lentiviral vectors carrying an enhanced synapsin I promoter.

    abstract:BACKGROUND:Certain inherited progressive neurodegenerative disorders, such as spinocerebellar ataxia (SCA), affect neurons in large areas of the central nervous system (CNS). The selective expression of disease-causing and therapeutic genes in susceptible regions and cell types is critical for the generation of animal ...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2013.12.004

    authors: Matsuzaki Y,Oue M,Hirai H

    更新日期:2014-02-15 00:00:00

  • RCA-I lectin histochemistry after trypsinisation enables the identification of microglial cells in thin paraffin sections of the mouse brain.

    abstract::A biotinylated lectin from Ricinus communis (RCA-I) and avidin-biotin-horseradish peroxidase complex (ABC) were used to identify microglial cells in 3-microns-thick sections of formalin-fixed paraffin-embedded brains of adult mice required for quantitative cell kinetic studies. In 3-microns-thick sections of the mouse...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/0165-0270(93)90034-o

    authors: Hauke C,Korr H

    更新日期:1993-12-01 00:00:00

  • Realistic simulation of extracellular recordings.

    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

    authors: Martinez J,Pedreira C,Ison MJ,Quian Quiroga R

    更新日期:2009-11-15 00:00:00

  • Chromatin immunoprecipitation and gene expression analysis of neuronal subtypes after fluorescence activated cell sorting.

    abstract:BACKGROUND:With advances in cell capture, gene expression can now be studied in neuronal subtypes and single cells; however, studying epigenetic mechanisms that underlie these changes presents challenges. Moreover, chromatin immunoprecipitation (ChIP) protocols optimized for low cell number do not adequately address te...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2016.02.006

    authors: Finegersh A,Homanics GE

    更新日期:2016-04-01 00:00:00

  • Neuromuscular electrical stimulation of the hindlimb muscles for movement therapy in a rodent model.

    abstract::Neuromuscular electrical stimulation (NMES) can provide functional movements in people after central nervous system injury. The neuroplastic effects of long-term NMES-induced repetitive limb movement are not well understood. A rodent model of neurotrauma in which NMES can be implemented may be effective for such inves...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2008.09.015

    authors: Ichihara K,Venkatasubramanian G,Abbas JJ,Jung R

    更新日期:2009-01-30 00:00:00

  • Performance variation in motor imagery brain-computer interface: a brief review.

    abstract::Brain-computer interface (BCI) technology has attracted significant attention over recent decades, and has made remarkable progress. However, BCI still faces a critical hurdle, in that performance varies greatly across and even within subjects, an obstacle that degrades the reliability of BCI systems. Understanding th...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章,评审

    doi:10.1016/j.jneumeth.2015.01.033

    authors: Ahn M,Jun SC

    更新日期:2015-03-30 00:00:00

  • Identification and investigation of mitochondria lacking cytochrome c oxidase activity in axons.

    abstract::Mitochondrial defects have been implicated in the degeneration of axons in a number of CNS disorders, including multiple sclerosis. Uniquely, mitochondria harbor the only non-nuclear DNA (mitochondrial DNA or mtDNA), which encodes functionally important subunits of the respiratory chain. The pattern of mitochondrial r...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2010.07.022

    authors: Zambonin J,Engeham S,Campbell GR,Ziabreva I,Beadle NE,Taylor RW,Mahad DJ

    更新日期:2010-09-30 00:00:00

  • Automated detection and analysis of foraging behavior in Caenorhabditis elegans.

    abstract::Foraging is a rapid, side-to-side movement of the nose generated by Caenorhabditis elegans as it explores its environment. In this paper, we present an automated method to detect and analyze foraging behavior of C. elegans in a video sequence. Several morphological image-processing methods are used to locate the preci...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2008.01.027

    authors: Huang KM,Cosman P,Schafer WR

    更新日期:2008-06-15 00:00:00

  • Linear inverse filtering improves spatial separation of nonlinear brain dynamics: a simulation study.

    abstract::We examined topographic variations in nonlinear measures based on scalp voltages, which were generated by two simulated current dipoles each placed in a different hemisphere of a spherical volume conductor (three-shell model). Dipole dynamics were that of a three-torus and the x-component of the Lorenz-system and scal...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/s0165-0270(00)00188-6

    authors: Fell J,Hauk O,Hinrichs H

    更新日期:2000-05-15 00:00:00

  • A perfused in vitro preparation of hypothalamus for electrophysiological studies on neurosecretory neurons.

    abstract::A technique for the preparation and recording of mammalian supraoptic nucleus neurosecretory neurons maintained in vitro through perfusion of the anterior cerebral artery is described. A slice of basal diencephalon measuring approximately 8 X 8 X 2 mm that includes the pituitary stalk and medial preoptic-anterior peri...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/0165-0270(83)90002-x

    authors: Bourque CW,Renaud LP

    更新日期:1983-03-01 00:00:00

  • Increased throughput assays of locomotor dysfunction in Drosophila larvae.

    abstract::Larval locomotion is a sensitive readout of a range of nervous system deficits in Drosophila, and has been utilised to quantify modulation of the disease phenotype in models of human disease. Single larvae are typically analysed in series using manual quantification of parameters such as contraction rate, or grouped t...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2011.08.037

    authors: Sinadinos C,Cowan CM,Wyttenbach A,Mudher A

    更新日期:2012-01-30 00:00:00

  • Novel model of orthotopic U-87 MG glioblastoma resection in athymic nude mice.

    abstract::In vitro and in vivo models of experimental glioma are useful tools to gain a better understanding of glioblastoma (GBM) and to investigate novel treatment strategies. However, the majority of preclinical models focus on treating solid intracranial tumours, despite surgical resection being the mainstay in the standard...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2017.04.019

    authors: Bianco J,Bastiancich C,Joudiou N,Gallez B,des Rieux A,Danhier F

    更新日期:2017-06-01 00:00:00

  • The use of whole-mount preparations of nerves labelled with axonally transported radioactive proteins to study regeneration.

    abstract::A method for studying the process of regeneration in the frog sciatic nerve is presented. The outgrowth of sensory axons was determined by assaying the distribution of axonally transported radioactive proteins by fluorography of whole-mount nerve preparations. After labelling, the nerves were fixed at their in vivo le...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/0165-0270(86)90004-x

    authors: Edström A,Sjöberg J,Kanje M

    更新日期:1986-03-01 00:00:00

  • Functional magnetic resonance imaging (FMRI) of the human brain.

    abstract::Functional magnetic resonance imaging (FMRI) can provide detailed images of human brain that reflect localized changes in cerebral blood flow and oxygenation induced by sensory, motor, or cognitive tasks. This review presents methods for gradient-recalled echo-planar functional magnetic resonance imaging (FMRI). Also ...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章,评审

    doi:10.1016/0165-0270(94)90191-0

    authors: DeYoe EA,Bandettini P,Neitz J,Miller D,Winans P

    更新日期:1994-10-01 00:00:00

  • Three dimensional rendering of auditory neuronal responses: A novel illustration of receptive field across frequency, intensity & time domains.

    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

    authors: Qureshi F,Yan J

    更新日期:2020-05-15 00:00:00

  • A new video/computer method to measure the amount of overall movement in experimental animals (two-dimensional object-difference method).

    abstract::Evaluation of the amount of overall animal movement is important for investigations of motor control mechanisms in the central nervous system. We describe a new method to quantify overall free movements of an animal without any markers using a video camera and a personal computer equipped with a video-capture board. T...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/s0165-0270(99)00082-5

    authors: Hashimoto T,Izawa Y,Yokoyama H,Kato T,Moriizumi T

    更新日期:1999-09-15 00:00:00

  • Glass-plastic push-pull cannula system for regional perfusion of sites in the brain.

    abstract::A new non-metallic push-pull cannula device is described for the localized perfusion of brain sites in the rat or other animal. Comprised entirely of glass and plastic materials, the cannula is intended for usage in an experiment in which the presence in tissue of metal tubing for delivery of perfusate is methodologic...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/0165-0270(83)90129-2

    authors: Myers RD,Hepler JR

    更新日期:1983-10-01 00:00:00

  • Cryopreserved rat cortical cells develop functional neuronal networks on microelectrode arrays.

    abstract::Neurons growing on microelectrode arrays (MEAs) are promising tools to investigate principal neuronal network mechanisms and network responses to pharmaceutical substances. However, broad application of these tools, e.g. in pharmaceutical substance screening, requires neuronal cells that provide stable activity on MEA...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/s0165-0270(03)00186-9

    authors: Otto F,Görtz P,Fleischer W,Siebler M

    更新日期:2003-09-30 00:00:00

  • High-performance liquid chromatography/tandem mass spectrometric assay for the simultaneous measurement of dopamine, norepinephrine, 5-hydroxytryptamine and cocaine in biological samples.

    abstract::A rapid liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed for the measurement of dopamine (DA), 5-hydroxytryptamine (5HT) and norepinephrine (NE) in brain microdialysates. The assay has also been utilised for the simultaneous measurement of these neurotransmitters and cocaine in brain...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2004.03.021

    authors: Hows ME,Lacroix L,Heidbreder C,Organ AJ,Shah AJ

    更新日期:2004-09-30 00:00:00

  • Cerebellar neurotransmission in attention-deficit/hyperactivity disorder: does dopamine neurotransmission occur in the cerebellar vermis?

    abstract::Children and adolescents with attention-deficit/hyperactivity disorder (ADHD) have smaller cerebellar volumes, particularly in the posterior-inferior cerebellar vermis (lobules VIII-X). Functional activation of the human cerebellar vermis following stimulant administration has also been repeatedly demonstrated. There ...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2005.09.019

    authors: Glaser PE,Surgener SP,Grondin R,Gash CR,Palmer M,Castellanos FX,Gerhardt GA

    更新日期:2006-02-15 00:00:00

  • Long-term in vitro turtle preparation for the study of spinal organization.

    abstract::A method by which muscles, nerves, and a section of spinal cord from a turtle can be stably maintained in vitro for many days has been developed. Tests of the viability of central and neuromuscular synapses, and the histochemical properties of muscle fibers have indicated that the preparation is viable and shows littl...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/0165-0270(89)90029-0

    authors: Lennard PR,Boss V,Shirley KA

    更新日期:1989-08-01 00:00:00

  • Finite element analysis of thermal laser skin stimulation for a finer characterization of the nociceptive system.

    abstract::Thermal laser stimulation of the skin is an efficient exploratory tool to characterize the nociceptive system. In the present study, finite element simulations are done to calculate the intra-cutaneous spatio-temporal temperature profiles following the delivery of such laser stimuli. The proposed computer-aided modeli...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/j.jneumeth.2013.11.010

    authors: Marchandise E,Mouraux A,Plaghki L,Henrotte F

    更新日期:2014-02-15 00:00:00

  • An improved invertebrate synaptosomal preparation with cholinergic properties.

    abstract::Our laboratory has previously characterized a high affinity choline uptake system (HAChUS) in Limulus tissues and synaptosomes. We report here on the characterization of the HAChUS in synaptosomes prepared selectively from central nervous system tissues shown to be enriched for presumed cholinergic functions; namely t...

    journal_title:Journal of neuroscience methods

    pub_type: 杂志文章

    doi:10.1016/0165-0270(91)90166-w

    authors: Torrence-Campbell C,Gates H,Effiong U,Sweet S,Townsel JG

    更新日期:1991-07-01 00:00:00