| 1. |
- Miliotis, T., et al.
(författare)
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Development of silicon microstructures and thin-film MALDI target plates for automated proteomics sample identifications
- 2001
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Ingår i: Journal of Neuroscience Methods. - : Elsevier BV. - 1872-678X .- 0165-0270. ; 109:1, s. 41-46
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Tidskriftsartikel (refereegranskat)abstract
- Here we report on the development of a proteomic platform utilizing a piezoelectric flow-through dispensing unit made from silicon microstructures. The use of a novel surface coating, where matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI MS) targets were uniformly precoated with a thin film of matrix/nitrocellulose, made the sample preparation straightforward and enabled the enrichment and analysis of proteins at low levels in proteomics samples. We demonstrate this by analyzing excised spots in a biological sample originating from a human fetal fibroblast cell line that was subjected to 2D gel-electrophoresis. Furthermore, a sample deposition rate below 30 Hz results in an increased analyte density on the dispensed sample spot, rendering signal amplification. In general, the sensitivity for proteins and peptides can be enhanced 10-50 times compared to traditional MALDI sample preparation techniques. (C) 2001 Elsevier Science B.V. All rights reserved.
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| 2. |
- Bergquist, Jonas, et al.
(författare)
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Capillary electrophoresis with laser-induced fluorescence detection : a sensitive method for monitoring extracellular concentrations of amino acids in the periaqueductal grey matter.
- 1996
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Ingår i: Journal of Neuroscience Methods. - : Elsevier BV. - 0165-0270 .- 1872-678X. ; 65:1, s. 33-42
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Tidskriftsartikel (refereegranskat)abstract
- The use of capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) for the analysis of microdialysate samples from the periaqueductal grey matter (PAG) of freely moving rats is described. By employing 3-(4-carboxybenzoyl)-2-quinoline-carboxaldehyde (CBQCA) as a derivatization agent, we simultaneously monitored the concentrations of 8 amino acids (arginine, glutamine, valine, gamma-amino-n-butyric acid (GABA), alanine, glycine, glutamate, and aspartate), with nanomolar and subnanomolar detection limits. Two of the amino acids (GABA and glutamate) were analysed in parallel by conventional high-performance liquid chromatography (HPLC) in order to directly compare the two analytical methods. Other CE methods for analysis of microdialysate have been previously described, and this improved method offers greater sensitivity, ease of use, and the possibility to monitor several amino acids simultaneously. By using this technique together with an optimised form of microdialysis technique, the tiny sample consumption and the improved detection limits permit the detection of fast and transient transmitter changes.
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| 3. |
- Isaac, Giorgis, et al.
(författare)
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Analysis of phosphatidylcholine and sphingomyelin molecular species from brain extracts using capillary liquid chromatography electrospray ionization mass spectrometry
- 2003
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Ingår i: Journal of Neuroscience Methods. - 0165-0270 .- 1872-678X. ; 128:1-2, s. 111-119
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Tidskriftsartikel (refereegranskat)abstract
- One feature of complex lipids is that many subtypes of these molecules exist as a diverse mixture in a biological sample. Qualitative and quantitative analysis of these closely related molecules require sensitive and specific analytical methods to detect intact phospholipids (PL) and sphingomyelin (SM) species and to differentiate between them. Conventional analytical methods require laborious procedures including separation by column, argentation thin-layer chromatography or liquid chromatography (LC) after pre- or post-column derivatization. In the present work, a method based on reversed phase capillary LC coupled on-line to electrospray ionization mass spectrometry (LC/ESI/MS) has been developed to gather tools for lipidomic studies, i.e. the profiling of complex mixtures of lipids in small amounts of various cells and tissues. The LC/MS system used consisted of an LC pump in an isocratic elution, a reversed phase capillary column and a single quadrupole mass spectrometer operated in the positive ion mode. A successful separation of phosphatidylcholine (PC) and SM molecular species was obtained with a minimum detectable quantity (MDQ) in the low fmol range injected on column. The method was applied to human brain extracts. Furthermore, the extraction efficiencies of the traditional Folch method and pressurized fluid extraction (PFE) were compared using the human brain. It was found that the intensity of the PC and SM molecular species extracted by PFE is two times that of Folch.
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| 4. |
- Novikov, Lev N
(författare)
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Labeling of central projections of primary afferents in adult rats : a comparison between biotinylated dextran amine, neurobiotin and Phaseolus vulgaris-leucoagglutinin.
- 2001
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Ingår i: Journal of Neuroscience Methods. - 0165-0270 .- 1872-678X. ; 112:2, s. 145-154
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Tidskriftsartikel (refereegranskat)abstract
- The efficacy of anterograde labeling of the central projections of primary afferent fibers were compared between biotinylated dextran amine (BDA), neurobiotin (NB) and Phaseolus vulgaris-leucoagglutinin (PHA-L) after injections into the L5 or T13 dorsal root ganglia (DRGs) of adult rats. Excellent labeling was obtained with BDA, which visualized fibers with fine terminal boutons in the L5 and T13 spinal cord segments, Clarke's nucleus and the gracile nucleus. Rarely observed crossed projections to the gracile nucleus and L5 ventral horn of the contralateral side could also be distinguished. Even in the most successful experiments, however, BDA labeled only about one-third of the axons originating from the injected dorsal root ganglion. BDA was also efficient as transganglionic tracer after application to the transected sciatic nerve. NB produced no significant labeling of the L5 primary afferents, and was only moderately effective on the T13 level. PHA injections resulted in sparse terminal labeling of the T13 and L5 afferents. Thus, BDA is an effective tracer for long-range labeling of primary afferent projections in the spinal cord and brain stem. Since not all stem fibers become labeled, however, the method does not allow quantification of all axon branches and terminals arising from the injected DRGs.
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| 5. |
- Öhberg, Fredrik, 1969-, et al.
(författare)
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A neural network appoach to real-time spike discrimination during simultaneous recording from several multi-unit nerve filaments
- 1996
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Ingår i: Journal of Neuroscience Methods. - : Elsevier BV. - 0165-0270 .- 1872-678X. ; 64:2, s. 181-187
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Tidskriftsartikel (refereegranskat)abstract
- A multi-channel, real-time, unsupervised spike discriminator was developed in order to reconstruct single spike trains from several simultaneously recorded multi-unit nerve filaments. The program uses a Self Organising Map (SOM) algorithm for the classification of the spikes. In contrast to previous similar techniques, the described method is made for use on a PC, and the method may thus be implemented at relatively low cost. In order to test the accuracy of the program, a robustness test was performed, where noise with different RMS levels was superimposed on the spikes. Furthermore, the maximal classification rate was determined. The program is easy to use, since the only manual inputs needed are the voltage threshold for spike detection, and the number of units present in each recorded nerve filament.
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| 6. |
- Afzali, Maryam, et al.
(författare)
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The sensitivity of diffusion MRI to microstructural properties and experimental factors
- 2021
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Ingår i: Journal of Neuroscience Methods. - : ELSEVIER. - 0165-0270 .- 1872-678X. ; 347
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Forskningsöversikt (refereegranskat)abstract
- Diffusion MRI is a non-invasive technique to study brain microstructure. Differences in the microstructural properties of tissue, including size and anisotropy, can be represented in the signal if the appropriate method of acquisition is used. However, to depict the underlying properties, special care must be taken when designing the acquisition protocol as any changes in the procedure might impact on quantitative measurements. This work reviews state-of-the-art methods for studying brain microstructure using diffusion MRI and their sensitivity to microstructural differences and various experimental factors. Microstructural properties of the tissue at a micrometer scale can be linked to the diffusion signal at a millimeter-scale using modeling. In this paper, we first give an introduction to diffusion MRI and different encoding schemes. Then, signal representation-based methods and multi-compartment models are explained briefly. The sensitivity of the diffusion MRI signal to the microstructural components and the effects of curvedness of axonal trajectories on the diffusion signal are reviewed. Factors that impact on the quality (accuracy and precision) of derived metrics are then reviewed, including the impact of random noise, and variations in the acquisition parameters (i.e., number of sampled signals, b-value and number of acquisition shells). Finally, yet importantly, typical approaches to deal with experimental factors are depicted, including unbiased measures and harmonization. We conclude the review with some future directions and recommendations on this topic.
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| 7. |
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| 8. |
- Ahemaiti, Aikeremu, 1984, et al.
(författare)
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A multifunctional pipette for localized drug administration to brain slices
- 2013
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Ingår i: Journal of Neuroscience Methods. - : Elsevier BV. - 0165-0270 .- 1872-678X. ; 219:2, s. 292-296
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Tidskriftsartikel (refereegranskat)abstract
- We have developed a superfusion method utilizing an open-volume microfluidic device for administration of pharmacologically active substances to selected areas in brain slices with high spatio-temporal resolution. The method consists of a hydrodynamically confined flow of the active chemical compound, which locally stimulates neurons in brain slices, applied in conjunction with electrophysiological recording techniques to analyze the response. The microfluidic device, which is a novel free-standing multifunctional pipette, allows diverse superfusion experiments, such as testing the effects of different concentrations of drugs or drug candidates on neurons in different cell layers with high positional accuracy, affecting only a small number of cells. We demonstrate herein the use of the method with electrophysiological recordings of pyramidal cells in hippocampal and prefrontal cortex brain slices from rats, determine the dependence of electric responses on the distance of the superfusion device from the recording site, document a multifold gain in solution exchange time as compared to whole slice perfusion, and show that the device is able to store and deliver up to four solutions in a series. Localized solution delivery by means of open-volume microfluidic technology also reduces reagent consumption and tissue culture expenses significantly, while allowing more data to be collected from a single tissue slice, thus reducing the number of laboratory animals to be sacrificed for a study. (C) 2013 Elsevier B.V. All rights reserved.
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| 9. |
- Ahemaiti, Aikeremu, 1984, et al.
(författare)
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Spatial characterization of a multifunctional pipette for drug delivery in hippocampal brain slices
- 2015
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Ingår i: Journal of Neuroscience Methods. - : Elsevier BV. - 0165-0270 .- 1872-678X. ; 241, s. 132-136
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Tidskriftsartikel (refereegranskat)abstract
- Background: Among the various fluidic control technologies, microfluidic devices are becoming powerful tools for pharmacological studies using brain slices, since these devices overcome traditional limitations of conventional submerged slice chambers, leading to better spatiotemporal control over delivery of drugs to specific regions in the slices. However, microfluidic devices are not yet fully optimized for such studies. New method: We have recently developed a multifunctional pipette (MFP), a free standing hydrodynamically confined microfluidic device, which provides improved spatiotemporal control over drug delivery to biological tissues. Results: We demonstrate herein the ability of the MFP to selectively perfuse one dendritic layer in the CA1 region of hippocampus with CNQX, an AMPA receptor antagonist, while not affecting the other layers in this region. Our experiments also illustrate the essential role of hydrodynamic confinement in sharpening the spatial selectivity in brain slice experiments. Concentration-response measurements revealed that the ability of the MFP to control local drug concentration is comparable with that of whole slice perfusion, while in comparison the required amounts of active compounds can be reduced by several orders of magnitude. Comparison with existing method: The multifunctional pipette is applied with an angle, which, compared to other hydrodynamically confined microfluidic devices, provides more accessible space for other probing and imaging techniques. Conclusions: Using the MFP it will be possible to study selected regions of brain slices, integrated with various imaging and probing techniques, without affecting the other parts of the slices.
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| 10. |
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