Die Autoren möchten Caroline Hartley und Rebeccah Slater (Kinderklinik, University of Oxford, UK) bedanken für die kritische Überprüfung unserer Papier- und Walter Magerl (Abteilung für Neurophysiologie, Zentrum für Biomedizin und Medizintechnik Mannheim (CBTM), Universität Heidelberg, Deutschland) für die Unterstützung mit Technik und wissen.

Die Studie wurde von der zuständigen Ethik Komitee der Nordwestschweiz (EKNZ 2015-079) genehmigt und fundierte schriftliche Einwilligung der Eltern für die Teilnehmer wurde vor der Messung erreicht.
1. Vorbereitung
<ol><li>Stellen Sie sicher, dass das Kind ausgeglichen wird. Es sollte ruhig und ließ sich, ohne saugen Bewegungen während der Aufnahme wegen Bewegungsartefakte. Das Baby kann schlafen9sein.</li>
	<li>Messen Sie das Neugeborene Kopfumfang mit einem Maßband um die Größe des EEG Kappe zu definieren.</li>
	<li>Identifizieren Sie die aktive Elektrodenposition: Cz / Vertexposition durch markieren den Mittelpunkt zwischen Nasion und Inion und dem mittleren Punkt zwischen der linken und rechten Preaurical Punkt mit einem Haut-Marker-Bleistift. Für die Positionierung der Elektroden siehe auch Abbildung 1.</li>
	<li>Identifizieren Sie die Positionen der Masseelektrode: rechten Stirn (Fp2) und die Referenzelektrode: linke Warzenfortsatzes (A1).</li>
	<li>Reinigen Sie die Elektrode Websites (Cz, Fp2 und A1) mit Desinfektionsmittel mit einem Wattestäbchen. Trennen Sie bei viele Haare die Haare um die Kopfhaut zu visualisieren.</li>
	<li>Schrubben Sie vorsichtig die Elektrode Websites mit EEG Mattierungspaste, mit einem Wattestäbchen auf um die Impedanz zu senken.</li>
	<li>Setzen Sie die EEG-Kappe mit auf das Neugeborene Kopf angebrachten Elektroden.</li>
	<li>Injizieren Sie leitendes EEG-Gel in die Elektroden mit einer Spritze mit einer kurzen Nadel aus Kunststoff, um den Kontakt zwischen den Elektroden und der Kopfhaut zu optimieren.</li>
	<li>Passen Sie die Elektroden, bis die Impedanz unter 50 kΩ ist, wie in den Empfehlungen des Herstellers und veröffentlichten Leitlinien12beschrieben.</li>
	<li>Stellen Sie eine Kamera um das Neugeborene Mimik zu erfassen.</li>
	<li>Verbinden Sie die Flat-Tip-Sonde mit Kontakt Trigger-Gerät, die EEG-Aufnahme-Gerät befestigt ist. Wenn die Wohnung-Tip Sonde Nennkraft auf der Haut erreicht, erzeugt ein Triggersignal Kontakt Trigger-Gerät. Dieses Signal wird an den Computer, das EEG-Aufzeichnung mit einem Trigger-Mark-tagging.</li>
</ol>2. Messung
<ol><li>Wählen Sie einen Studiennamen zum Speichern der Daten.</li>
	<li>
Online-Band Pass Filter hinzufügen:
	<ol>
<li>Display-Filter: Niedrige cutoff-Filter: Frequenz: 1 Hz Hohen cutoff-Filter: Frequenz: 70 Hz Notch-Filter: 50 Hz Abtastrate: 2000 Hz</li>
	</ol>
</li>
	<li>Starten Sie das EEG und video-Aufzeichnung.</li>
	<li>Halten Sie das Neugeborene Rechte Hand in einer horizontalen Position. Der Reiz auch auf einer anderen Website angewendet werden kann, z.B. den Fuß, was zu längeren Latenz als von Hand führt, Einzelheiten finden Sie 3,7.</li>
	<li>Erste Platte Hintergrund EEG-Aktivität. Während das Neugeborene Hand gehalten wird, kommentieren Sie die EEG-Aufzeichnung manuell, um Zeiträume zu erfassen, wo keine Reize gelten und das Kind ruht.</li>
	<li>Führen Sie die erforderliche Menge an Flat-Tip Sonde Reize nach das Studiendesign auf das Neugeborene Rechte Hand. Inmitten unserer experimentellen Design, das die Anzahl der Flat-Tip Sonde Reize um 50. Achten Sie darauf, die Flat-Tip Sonde senkrecht auf das Neugeborene Hand verwenden, so dass die Spitze nicht beugen und die richtige Kraft aufgebracht. Führen Sie die Flat-Tip Sonde Reize mit einem Minimum an einem 2 bis 3 s inter-Stimulus-Intervall (ISI)6 , um Summierung zu vermeiden.</li>
	<li>Die Aufnahmen zu stoppen.</li>
	<li>Die experimentelle Einstellungsdetails zu dokumentieren.</li>
</ol>(3) Datenanalyse
<ol><li>Filtern Sie die Rohdaten EEG offline mit einem Hochpass-Filter bei 1 Hz und einem Tiefpass-Filter bei 30 Hz.</li>
	<li>Segmentieren Sie die Daten in Epochen der 1.500 ms (500 ms vor 1.000 MS nach dem Stimulus Onset).</li>
	<li>Führen Sie eine Basislinienkorrektur, das Pre-Stimulus-Intervall.</li>
	<li>Manuell ablehnen der EEG-Epochen mit Artefakten wie Bewegungsartefakte und Lärm, nach Sichtprüfung. Siehe hierzu auch den Tutorial Umriss: Https://sccn.ucsd.edu/wiki/Chapter_01:_Rejecting_Artifacts</li>
	<li>Durchschnitt der EEG-Epochen (für den Hintergrund und Stimulus Reaktion separat).</li>
	<li>Holzig-Filter die Daten mit einer maximalen Jitter von ± 50 ms im Zeitfenster 0 - 1000 ms post Reiz auftreten. Dies ermöglicht eine Latenz Unterschiede zwischen den Kindern. Dies gilt für den Hintergrund und Reiz Antworten separat.</li>
	<li>Projekt-Vorlage schädliche-evozierten Hirnaktivität, die in einem unabhängigen Dataset definiert wurde und wurde in beschrieben ausführlich an anderer Stelle11 auf die Daten, die das Ausmaß der schädlichen-evozierten Reaktion innerhalb jeder einzelnen Prüfung zu ermitteln. Die Vorlage beschreibt eine charakteristische Wellenform der schädlichen-evozierten Reaktion und das Ausmaß spiegelt die Menge dieser schädlichen-evozierten Reaktion innerhalb der einzelnen Studie. Hinweis: Das Zeitfenster, um die Vorlage zu projizieren ist abhängig vom Alter des Säuglings und die Impulse Position. Wenn Reize auf die Hand angewendet werden, dann ist das Zeitfenster von Interesse Posten 200-500 ms Impuls11. Wenn die Reize auf den Fuß angewendet werden, dann ist das Zeitfenster von Interesse post 400-700 ms Impuls11. Beachten Sie, dass diese Vorlage nur derzeit für Kleinkinder zwischen 34-43 Wochen Schwangerschaft bestätigt wurde. Für detaillierte Methoden verbunden mit der Verwendung dieser Vorlage und eine Diskussion über die Grenzen bitte siehe Hartley Et Al. 201711.</li>
	<li>Überprüfen Sie, dass das Ausmaß der Aktivität des Gehirns schädliche-evozierten, dass mithilfe der Vorlage deutlich den Reiz berechnet, im Vergleich mit im Hintergrund EEG verfolgt.</li>
	<li>Analysieren des Verhalten Gesichtsausdruck danach (Kamera ist verknüpft mit dem EEG Aufzeichnung) mit der neonatalen Gesichts Coding System2,13.</li>
</ol>

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V., Johnston, C. C., Craig, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+facial+and+cry+responses+to+invasive+and+non-invasive+procedures.">Neonatal facial and cry responses to invasive and non-invasive procedures.</a> Pain. 42 (3), 295-305 (1990).</li><li>van den Broeke, E. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterizing+pinprick-evoked+brain+potentials+before+and+after+experimentally+induced+secondary+hyperalgesia.">Characterizing pinprick-evoked brain potentials before and after experimentally induced secondary hyperalgesia.</a> J Neurophysiol. 114 (5), 2672-2681 (2015).</li><li>Verriotis, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+activity+evoked+by+inoculation+needle+prick+in+infants+up+to+one-year+old.">Cortical activity evoked by inoculation needle prick in infants up to one-year old.</a> Pain. 156 (2), 222-230 (2015).</li><li>Rolke, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+sensory+testing+in+the+German+Research+Network+on+Neuropathic+Pain+(DFNS):+standardized+protocol+and+reference+values.">Quantitative sensory testing in the German Research Network on Neuropathic Pain (DFNS): standardized protocol and reference values.</a> Pain. 123 (3), 231-243 (2006).</li><li>Moultrie, F., Slater, R., Hartley, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+the+treatment+of+infant+pain.">Improving the treatment of infant pain.</a> Curr Opin Support Palliat Care. 11 (2), 112-117 (2017).</li><li>Hu, L., Zhang, Z. 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Die Autoren haben nichts preisgeben.

Der hier vorgestellte Ansatz zeigt wie schädliche hervorgerufen Gehirn Aktivität beim Neugeborenen in objektiver Weise mit EEG-Aufzeichnung und Flat-Tip Sonde Stimulatoren experimentelle Schmerzreize anwenden gemessen werden. Diese Technik kann in verschiedenen klinischen Situationen verwendet werden, Nozizeption, z. B. in non-verbale Personen wie Neugeborene zu erkennen. Die vollständige Studie kann innerhalb von 15 Minuten, einschließlich platzieren das Baby, Festlegung, Ausarbeitung und Befestigung der El...

Schmerz ist eine unangenehme sensorische und emotionale Erfahrung verbunden mit tatsächlicher oder potenzieller Gewebeschädigung oder in Bezug auf solche Schäden1beschrieben. Die Unfähigkeit, verbal zu kommunizieren, negiert nicht die Möglichkeit, dass ein Individuum Schmerzen erlebt aber es sehr schwierig macht zu beurteilen, schmerzlindernde Behandlung, zum Beispiel bei Neugeborenen2. Verschiedene Verhaltens- und physiologischen Indikatoren sind verwendet, um Schmerzen bei non-verbale Patienten beurteilen. Im Laufe der Jahre wurden verschiedene Skalen entwickelt, die Wahl hängt von der Art des Reizes, gestational Alter und das Umfeld, in dem der Neugeborenen Embeded3,4,5. Diese Schmerzen Bewertungsinstrumente entweder auf die Rater Auslegung verlassen oder sie fordern sekundären physiologischen Indikatoren.
In diesem Video präsentieren wir Ihnen eine Methode zur Bestimmung der Nozizeption elektrophysiologische Aufnahmen optimiert für den Einsatz bei Neugeborenen. Nozizeption ist definiert als die neuronalen Prozess der Codierung Schmerzreize. So ist die Quantifizierung der Nozizeption eine elegante und objektive Methode zur Bestimmung des neuronalen Eingangs in eine non-verbale Person. Darüber hinaus ist kortikale Aktivität erkannt durch Elektroenzephalographie (EEG) korreliert mit der Intensität der schädlichen Ereignisse5,6.
Die hier vorgestellte Methode kombiniert mit Schmerzreize produziert durch eine mechanische Stimulation mit einer flachen Spitze Sonde, auch genannt eine Pinprick7, das ist nicht Haut-Abbau und verursacht keine Verhaltensstörungen not6EEG-Aufzeichnung. Es hat sich gezeigt, dass Nozizeption folgenden Stimulation interpunktieren ist vorwiegend vermittelt durch Aδ-Fasern, ist nicht notwendig eine Haut brechen Läsion8 und das Ausmaß der nozizeptiven-spezifischen potenziellen nicht abhängig von ist Zustand9schlafen. Die Sonde ist von Eltern auch gut angenommen, bei der Anwendung dieser Methode in einer Studie Umgebung mit Neugeborenen. Die Sonde ist elektronisch mit der EEG-Aufnahme-System ermöglicht die EEG-Aufzeichnung genau gekennzeichnet werden, wenn die Sonde Kontakt mit der Haut verbunden. Dies vereinfacht den Prozess der Zeit sperren und ist die Voraussetzung für alle nachfolgenden EEG-Analysen. Um die Vorbereitungszeit der Säugling EEG-Aufzeichnung zu minimieren haben wir einen veränderten internationalen 10/20 Platzierung Elektrodensystem wo haben wir die Anzahl der Elektroden auf die Mindestanforderung von drei Elektroden (Abbildung 1) reduziert. Die zentrale Stützpunkt Cz Elektrode, wo die schädliche-evozierten Hirnaktivität maximal9,10,11 ist, wurde zusammen mit einem Verweis und eine Masseelektrode eingesetzt.

<table>
	<tbody>
		<tr>
			<td>Easycap</td>
			<td>EASYCAP GmbH</td>
			<td>AC-32-C</td>
			<td>EEG caps for infants sizes 34 and 36</td>
		</tr>
		<tr>
			<td>actiCAP</td>
			<td>Brain Products GmbH</td>
			<td>BP-04243-SIG</td>
			<td>active electrodes</td>
		</tr>
		<tr>
			<td>ImpBox</td>
			<td>Brain Products GmbH</td>
			<td></td>
			<td>impedance measurement</td>
		</tr>
		<tr>
			<td>V-Amp</td>
			<td>Brain Products GmbH</td>
			<td></td>
			<td>EEG recording device</td>
		</tr>
		<tr>
			<td>Contact trigger for pinprick stimulation</td>
			<td>MRC Systems GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PinPrick stimulator set</td>
			<td>MRC Systems GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>EEG prepping paste</td>
			<td>USB Pharmacy</td>
			<td></td>
			<td>contains sodium chloride, pumice stone, propylene glycol</td>
		</tr>
		<tr>
			<td>SuperVisc</td>
			<td>EASYCAP GmbH</td>
			<td></td>
			<td>Electrolyte-Gel for active electrodes</td>
		</tr>
		<tr>
			<td>Brain Vision Recorder</td>
			<td>Brain Products GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Brain Vision Analyzer</td>
			<td>Brain Products GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MATLAB using EEGLAB
			
			
			
			
			
			</td>
			<td>Swartz Center for Computational Neuroscience, University of California San Diego</td>
			<td></td>
			<td>For EEG processing, including averaging of all EEG epochs
			
			
			
			</td>
		</tr>
	</tbody>
</table>

electrophysiological measurement of noxious-evoked brain activity in neonates using a flat-tip probe coupled to electroencephalography

Schmerz ist eine unangenehme sensorische und emotionale Erfahrung. Bei non-verbale Patienten ist es sehr schwierig, Schmerzen, auch mit Schmerzen-Bewertungs-Tools zu messen. Diese Werkzeuge sind subjektiv oder sekundären physiologischen Indikatoren, die auch gewisse Einschränkungen haben, besonders, wenn die Wirksamkeit der Analgesie erkunden zu bestimmen. Da kortikalen Verarbeitung für Schmerzwahrnehmung unverzichtbar ist, können Gehirn Aktivität Maßnahmen eine sinnvolle Vorgehensweise zur Beurteilung von Schmerzen bei Säuglingen. Hier präsentieren wir eine Methode zur Bestimmung der Nozizeption mit elektrophysiologischen Gehirn Tätigkeit Aufnahmen optimiert für den Einsatz bei Neugeborenen. Um hoch standardisierte und reproduzierbare Schmerzreize erzeugen wir mechanische Stimulation mit einer Flat-Tip-Sonde, z.B.PinPrick, die ist nicht Haut-Abbau und verursacht keine Verhaltensstörungen not angewendet. Das schädliche evozierte Potential ermöglicht die objektive Messung der Nozizeption in non-verbale Patienten. Diese Methode kann bei Neugeborenen bereits 34 Wochen gestational Alter verwendet werden. Darüber hinaus kann es in verschiedenen Situationen wie die Messung der Wirksamkeit von analgetische oder betäubende Medikamente angewendet werden.

Abbildung 1 zeigt die schematische Darstellung der Positionierung mithilfe der veränderten internationalen 10/20 Platzierung Elektrodensystem Elektrode. Abbildung 2 zeigt die EEG-Aktivität aufgezeichnet kurz vor und nach Anwendung einer einzigen schädliche Stimulus mit einer Flat-Tip Sonde mit 32 mN Kraft Stimulation aufgetreten wie beschrieben in das Protokoll, das Neugeborene Rechte Hand (Abbildung 2A</s...

Watch this Scientific Journal Video about Electrophysiological Measurement of Noxious-evoked Brain Activity in Neonates Using a Flat-tip Probe Coupled to Electroencephalography at JoVE.com

Electrophysiological Measurement of Noxious-evoked Brain Activity in Neonates Using a Flat-tip Probe Coupled to Electroencephalography

Elektrophysiologische Messungen der schädlichen-evozierten Hirnaktivität beim Neugeborenen mit einem Flat-Tip Sonde gekoppelt, Elektroenzephalographie

Schmerzen bei non-verbale Patienten zu messen, ist eine Herausforderung. In dieser Studie verbinden wir EEG-Aufzeichnung mit der Stimulation mit einer Flat-Tip-Sonde, um schädliche-evozierten Hirnaktivität in objektiver Weise zu erkennen.

Pain is an unpleasant sensory and emotional experience. In non-verbal patients, it is very difficult to measure pain, even with pain assessment tools. ...

electrophysiological-measurement-noxious-evoked-brain-activity

Research

JoVE Journal

Medicine

6.5K Aufrufe.  University of Basel Children's Hospital. Schmerzen bei non-verbale Patienten zu messen, ist eine Herausforderung. In dieser Studie verbinden wir EEG-Aufzeichnung mit der Stimulation mit einer Flat-Tip-Sonde, um schädliche-evozierten Hirnaktivität in objektiver Weise zu erkennen.

Serie: Electrophysiological Measurement of Noxious-evoked Brain Activity in Neonates Using a Flat-tip Probe Coupled to Electroencephalography

Segmentation and Measurement of Fat Volumes in Murine Obesity Models Using X-ray Computed Tomography

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are associated with obesity, though the precise underlying mechanisms that contribute to the disease are currently being delineated.2,3 Several small animal models of obesity have been developed and are employed in a variety of studies.4 A critical component to these experiments involves the collection of regional and/or total animal fat content data under varied conditions.
Traditional experimental methods available for measuring fat content in small animal models of obesity include invasive (e.g. ex vivo measurement of fat deposits) and non-invasive (e.g. Dual Energy X-ray Absorptiometry (DEXA), or Magnetic Resonance (MR)) protocols, each of which presents relative trade-offs. Current invasive methods for measuring fat content may provide details for organ and region specific fat distribution, but sacrificing the subjects will preclude longitudinal assessments. Conversely, current non-invasive strategies provide limited details for organ and region specific fat distribution, but enable valuable longitudinal assessment. With the advent of dedicated small animal X-ray computed tomography (CT) systems and customized analytical procedures, both organ and region specific analysis of fat distribution and longitudinal profiling may be possible. Recent reports have validated the use of CT for in vivo longitudinal imaging of adiposity in living mice.5,6 Here we provide a modified method that allows for fat/total volume measurement, analysis and visualization utilizing the Carestream Molecular Imaging Albira CT system in conjunction with PMOD and Volview software packages.

Fat content analysis is routinely conducted in studies utilizing murine obesity models. Emerging methods in small animal CT imaging and analysis are providing for longitudinal detail rich fat content analysis. Here we detail step by step procedures for performing small animal CT imaging, analysis, and visualization.

Segmentierung und Messung von FAT-Volumes in Murine Obesity Modellen mithilfe von Röntgen-Computertomographie

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are ...

Forschung

Medizin

Epidural Intracranial Pressure Measurement in Rats Using a Fiber-optic Pressure Transducer

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and cardiac arrest. This elevation may result in further neurological injury, in the form of transtentorial herniation1,2,3,4, midbrain compression, neurological deficit or increased cerebral infarct2,4. Current therapies are often inadequate to control elevated ICP in the clinical setting5,6,7 . Thus there is a need for accurate methods of ICP measurement in animal models to further our understanding of the basic mechanisms and to develop new treatments for elevated ICP.
In both the clinical and experimental setting ICP cannot be estimated without direct measurement. Several methods of ICP catheter insertion currently exist. Of these the intraventricular catheter has become the clinical 'gold standard' of ICP measurement in humans8. This method involves the partial removal of skull and the instrumentation of the catheter through brain tissue. Consequently, intraventricular catheters have an infection rate of 6-11%9. For this reason, subdural and epidural cannulations have become the preferred methods in animal models of ischemic injury. 
Various ICP measurement techniques have been adapted for animal models, and of these, fluid-filled telemetry catheters10 and solid state catheters are the most frequently used11,12,13,14,15. The fluid-filled systems are prone to developing air bubbles in the line, resulting in false ICP readings. Solid state probes avoid this problem (Figure 1). An additional problem is fitting catheters under the skull or into the ventricles without causing any brain injury that might alter the experimental outcomes. Therefore, we have developed a method that places an ICP catheter contiguous with the epidural space, but avoids the need to insert it between skull and brain. 
An optic fibre pressure catheter (420LP, SAMBA Sensors, Sweden) was used to measure ICP at the epidural location because the location of the pressure sensor (at the very tip of the catheter) was found to produce a high fidelity ICP signal in this model. There are other manufacturers of similar optic fibre technologies13 that may be used with our methodology. Alternative solid state catheters, which have the pressure sensor located at the side of the catheter tip, would not be appropriate for this model as the signal would be dampened by the presence of the monitoring screw. 
Here, we present a relatively simple and accurate method to measure ICP. This method can be used across a wide range of ICP related animal models.

A novel technique to record the pressures within the skull is described. The minimally invasive method uses a fibre-optic pressure sensing system to accurately measure intracranial pressure (ICP) in anaesthetized rats without causing significant brain trauma. The technique may be used in a wide range of experimental models.

Epidurale Intrakranielle Druckmessung in Ratten mit einem LWL-Druckaufnehmer

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and ...

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

Die Messung und Behandlung von Unterdrückung in Amblyopie

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

State of the Art Cranial Ultrasound Imaging in Neonates

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a patient is unstable. In addition it is radiation-free and allows serial imaging. CUS possibilities have steadily expanded. However, in many neonatal intensive care units, these possibilities are not optimally used. We present a comprehensive approach for neonatal CUS, focusing on optimal settings, different probes, multiple acoustic windows and Doppler techniques. This approach is suited for both routine clinical practice and research purposes. In a live demonstration, we show how this technique is performed in the neonatal intensive care unit. Using optimal settings and probes allows for better imaging quality and improves the diagnostic value of CUS in experienced hands. Traditionally, images are obtained through the anterior fontanel. Use of supplemental acoustic windows (lambdoid, mastoid, and lateral fontanels) improves detection of brain injury. Adding Doppler studies allows screening of patency of large intracranial arteries and veins. Flow velocities and indices can be obtained. Doppler CUS offers the possibility of detecting cerebral sinovenous thrombosis at an early stage, creating a window for therapeutic intervention prior to thrombosis-induced tissue damage. Equipment, data storage and safety aspects are also addressed.

Cranial ultrasound (CUS) is a valuable tool for brain imaging in critically ill neonates. This video shows a comprehensive approach for neonatal (Doppler) CUS for both clinical and research purposes, including a bedside demonstration of the technique.

State of the Art Hirn Ultraschall-Bildgebung bei Neugeborenen

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a ...

Molecular Probe Optimization to Determine Cell Mortality in a Photosynthetic Organism (Microcystis aeruginosa) Using Flow Cytometry

Microbial subpopulations in field and laboratory studies have been shown to display high heterogeneity in morphological and physiological parameters. Determining the real time state of a microbial cell goes beyond live or dead categories, as microbes can exist in a dormant state, whereby cell division and metabolic activities are reduced. Given the need for detection and quantification of microbes, flow cytometry (FCM) with molecular probes provides a rapid and accurate method to help determine overall population viability. By using SYTOX Green and SYTOX Orange in the model cyanobacteria Microcystis aeruginosa to detect membrane integrity, we develop a transferable method for rapid indication of single cell mortality. The molecular probes used within this journal will be referred to as green or orange nucleic acid probes respectively (although there are other products with similar excitation and emission wavelengths that have a comparable modes of action, we specifically refer to the fore mentioned probes). Protocols using molecular probes vary between species, differing principally in concentration and incubation times. Following this protocol set out on M.aeruginosa the green nucleic acid probe was optimized at concentrations of 0.5 &#181;M after 30 min of incubation and the orange nucleic acid probe at 1 &#181;M after 10 min. In both probes concentrations less than the stated optimal led to an under reporting of cells with membrane damage. Conversely, 5 &#181;M concentrations and higher in both probes exhibited a type of non-specific staining, whereby &#39;live&#39; cells produced a target fluorescence, leading to an over representation of &#39;non-viable&#39; cell numbers. The positive controls (heat-killed) provided testable dead biomass, although the appropriateness of control generation remains subject to debate. By demonstrating a logical sequence of steps for optimizing the green and orange nucleic acid probes we demonstrate how to create a protocol that can be used to analyse cyanobacterial physiological state effectively.

Molecular Probe Optimization to Determine Cell Mortality in a Photosynthetic Organism Microcystis aeruginosa Using Flow Cytometry

Microbial populations contain substantial cell heterogeneity, which can dictate overall behavior. Molecular probe analysis through flow cytometry can determine physiological states of cells, however its application varies between species. This study provides a protocol to accurately determine cell mortality within a cyanobacterium population, without underestimating or recording false positive results.

Molecular Probe-Optimierung, um Zellmortalität in einem photosynthetische Organismen Bestimmen (<em&gt; Microcystis aeruginosa</em&gt;) Mittels Durchflusszytometrie

Microbial subpopulations in field and laboratory studies have been shown to display high heterogeneity in morphological and physiological parameters. ...

Transcutaneous Microcirculatory Imaging in Preterm Neonates

Microcirculatory imaging (MI) is a relatively new research tool mainly used in the intensive care setting. MI provides a clear view of the smallest capillaries, arterioles and venules. The magnifying effect visualizes the flow pattern of erythrocytes through these vessels.
It's non-invasive character makes it suitable to apply in (preterm) neonates, even in cardiorespiratory unstable patients. In adults and children, MI is mainly performed sublingually, but this is not possible in preterm infants as these cannot cooperate and the size of the probe is problematic. In preterm infants, MI is therefore performed transcutaneously. Their thin skin makes it possible to obtain high quality images of peripheral microcirculation. 
In this manuscript we will demonstrate the method of transcutaneous MI in preterm infants. We will focus on the different techniques and provide tips to optimize image quality. The highlights of software settings, safety and offline analysis are also addressed.

Microcirculatory imaging (MI) is used to monitor peripheral perfusion in critically ill or preterm neonates. This manuscript and video demonstrates the optimal approach for obtaining high-quality images.

Transkutane Mikrozirkulatorische Imaging in Frühgeborenen

Microcirculatory imaging (MI) is a relatively new research tool mainly used in the intensive care setting. MI provides a clear view of the smallest ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including Parkinson's and Huntington's can disrupt it. People with Parkinson's disease, for example, tend to have increased saccadic latencies. Consequently, the quantitative measurement of saccadic eye movements has received considerable attention as a potential biomarker for neurodegenerative conditions. A lot more can be learned about the brain in both health and disease by observing what happens to eye movements when the function of specific brain areas is perturbed. Deep brain stimulation is a surgical intervention used for the management of a range of neurological conditions including Parkinson's disease, in which stimulating electrodes are placed in specific brain areas including several sites in the basal ganglia. Eye movement measurements can then be made with the stimulator systems both off and on and the results compared. With suitable experimental design, this approach can be used to study the pathophysiology of the disease being treated, the mechanism by which DBS exerts it beneficial effects, and even aspects of normal neurophysiology.

This paper describes the use of quantitative measurement of eye movements in conjunction with stimulation of focal areas of the deep brain in order to study physiology, pathophysiology, and the mechanisms of deep brain stimulation.

Mit Saccadometry mit Deep Brain Stimulation Normale und Pathologische Funktion des Gehirns zu studieren

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...

Imaging Metals in Brain Tissue by Laser Ablation - Inductively Coupled Plasma - Mass Spectrometry (LA-ICP-MS)

Metals are found ubiquitously throughout an organism, with their biological role dictated by both their chemical reactivity and abundance within a specific anatomical region. Within the brain, metals have a highly compartmentalized distribution, depending on the primary function they play within the central nervous system. Imaging the spatial distribution of metals has provided unique insight into the biochemical architecture of the brain, allowing direct correlation between neuroanatomical regions and their known function with regard to metal-dependent processes. In addition, several age-related neurological disorders feature disrupted metal homeostasis, which is often confined to small regions of the brain that are otherwise difficult to analyze. Here, we describe a comprehensive method for quantitatively imaging metals in the mouse brain, using laser ablation - inductively coupled plasma - mass spectrometry (LA-ICP-MS) and specially designed image processing software. Focusing on iron, copper and zinc, which are three of the most abundant and disease-relevant metals within the brain, we describe the essential steps in sample preparation, analysis, quantitative measurements and image processing to produce maps of metal distribution within the low micrometer resolution range. This technique, applicable to any cut tissue section, is capable of demonstrating the highly variable distribution of metals within an organ or system, and can be used to identify changes in metal homeostasis and absolute levels within fine anatomical structures.

Imaging Metals in Brain Tissue by Laser Ablation - Inductively Coupled Plasma - Mass Spectrometry LA-ICP-MS

Quantitatively mapping metals in tissue by laser ablation - inductively coupled plasma - mass spectrometry (LA-ICP-MS) is a sensitive analytical technique that can provide new insight into how metals participate in normal function and disease processes. Here, we describe a protocol for quantitatively imaging metals in thin sections of mouse neurological tissue.

Imaging Metals im Hirngewebe durch Laserablation - Inductively Coupled Plasma - Mass Spectrometry (LA-ICP-MS)

Metals are found ubiquitously throughout an organism, with their biological role dictated by both their chemical reactivity and abundance within a ...

Absolute Quantification of A&#946;1-42 in CSF Using a Mass Spectrometric Reference Measurement Procedure

Alzheimer's disease (AD) is the most common neurodegenerative disease among the elderly and accounts for 60-80% of all cases of dementia. Currently, the diagnosis of AD is based on cognitive tests and mental state exams, but the peptide amyloid-beta (A&#946;) in cerebrospinal fluid (CSF) is increasingly used in clinical trials and settings. As for most protein and peptide biomarkers, quantification is performed using antibody-based techniques, such as enzyme-linked immunosorbent assay (ELISA). However, intra- and inter-laboratory variability in these assays hamper its use as a diagnostic marker in clinical routine.
An antibody-independent Reference Measurement Procedure (RMP) was developed based on solid-phase extraction (SPE) and liquid chromatography (LC)-tandem mass spectrometry (MS/MS), where stable, isotope-labeled A&#946; peptides were used as internal standards, enabling absolute quantification. A high-resolution quadrupole-orbitrap hybrid instrument was used for the measurements. The method allows for the quantification of CSF A&#946;1-42 between 150-4,000 pg/mL.

A reference measurement procedure for the absolute quantification of A&#946;1-42 in human CSF based on solid-phase extraction and liquid chromatography tandem mass spectrometry is described.

Absolute Quantifizierung von Aß<sub&gt; 1-42</sub&gt; In CSF eine massenspektrometrische Referenzmessverfahren verwenden

Alzheimer's disease (AD) is the most common neurodegenerative disease among the elderly and accounts for 60-80% of all cases of dementia. Currently, the ...

Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated measurements over weeks or months of accessory respiratory muscle activity while simultaneously measuring ventilation in a non-anesthetized, freely behaving mouse. The technique includes the surgical implantation of a radio transmitter and the insertion of electrode leads into the scalene and trapezius muscles to measure the electromyogram activity of these inspiratory muscles. Ventilation is measured by whole-body plethysmography, and animal movement is assessed by video and is synchronized with electromyogram activity. Measurements of muscle activity and ventilation in a mouse model of amyotrophic lateral sclerosis are presented to show how this tool can be used to investigate how respiratory muscle activity changes over time and to assess the impact of muscle activity on ventilation. The described methods can easily be adapted to measure the activity of other muscles or to assess accessory respiratory muscle activity in additional mouse models of disease or injury.

This paper introduces a method for repeated measurements of ventilation and respiratory muscle activity in a freely behaving amyotrophic lateral sclerosis (ALS) mouse model throughout disease progression with whole-body plethysmography and electromyography via an implanted telemetry device.

Wiederholte Messung der Atemmuskelaktivität und Ventilation in Mausmodellen von neuromuskulären Erkrankungen

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated ...