الكتاب يود أن يعترف هارتلي كارولين وسلاتر ريبيككة (قسم طب الأطفال، جامعة أكسفورد، المملكة المتحدة) لاستعراض الحرجة لدينا ورقة ووالتر ماجيرل (قسم من العصبية، ومركز للطب الحيوي والتكنولوجيا الطبية مانهايم (كبتم)، جامعة هايدلبرغ، ألمانيا) لدعم لنا بالمعدات التقنية والمعرفة.

تمت الموافقة على الدراسة المختصة أخلاقيات اللجنة من شمال غرب سويسرا (أكنز عام 2015-079) وتم الحصول على موافقة الوالدين خطيا مستنيرة للمشارك قبل أخذ القياس.
1-إعداد
<ol><li>تأكد من أن تتم تسوية الطفل. ينبغي أن يكون هادئة ومستقرة، دون مص الاقتراحات أثناء التسجيل بسبب حركة المصنوعات اليدوية. يمكن أن يكون الطفل نائماً9.</li>
	<li>قياس محيط الرأس للوليد مع شريط قياس لتحديد حجم الغطاء EEG.</li>
	<li>تحديد موقف القطب النشط: تشيكوسلوفاكيا/نقطة ذروة الموقف بوضع علامة النقطة المتوسطة بين ناسيون وإينيون ونقطة الوسط بين برياوريكال اليسرى واليمنى بقلم ماركر جلد. لتحديد المواقع الأقطاب انظر أيضا الشكل 1.</li>
	<li>تحديد مواقف القطب الأرضي: جبهته اليمنى (Fp2)، ومسرى المرجع: الخشاء اليسرى (A1).</li>
	<li>تنظيف المواقع قطب كهربائي (تشيكوسلوفاكيا و Fp2 و A1) مع مسحه القطن باستخدام مطهر. في حالة كثير من الشعر، فصل الشعر لتصور فروة الرأس.</li>
	<li>بلطف فرك مواقع القطب مع التخطيط الدماغي الاستعداد اللصق، استخدام مسحه القطن، إلى انخفاض المقاومة.</li>
	<li>وضع سقف التخطيط الدماغي مع أقطاب كهربائية تعلق على رأسه للوليد.</li>
	<li>حقن موصلة EEG جل أقطاب كهربائية باستخدام حقنه بإبرة بلاستيكية قصيرة لتحسين الاتصال بين الأقطاب وفروة الرأس.</li>
	<li>ضبط الأقطاب حتى المقاومة أدناه 50 kΩ كما ورد في توصيات الشركة المصنعة وفي المبادئ التوجيهية المنشورة12.</li>
	<li>ضع كاميرا لتسجيل تعابير الوجه للوليد.</li>
	<li>قم بتوصيل المجس tip مسطحة-الجهاز الاتصال المشغل، والذي هو ثابت لجهاز تسجيل EEG. عندما يصل المسبار مسطحة-تلميح إلى القوة الاسمية على الجلد، تولد إشارة الزناد الزناد اتصال الجهاز. يتم إرسال هذه الإشارات إلى الكمبيوتر، ووضع علامات EEG تسجيل علامة على الزناد.</li>
</ol>2-قياس
<ol><li>حدد اسم دراسة لتخزين البيانات.</li>
	<li>
إضافة عوامل تصفية تمرير النطاق على الإنترنت:
	<ol>
<li>عرض عوامل التصفية: تصفية قطع منخفضة: التردد: 1 هرتز تصفية قطع عالية: التردد: هرتز 70 تصفية الشق: 50 هرتز معدل العينات: 2000 هرتز</li>
	</ol>
</li>
	<li>بدء التخطيط الدماغي وتسجيل الفيديو.</li>
	<li>عقد اليد اليمنى للوليد في وضع أفقي. يمكن أيضا تطبيق الحافز في موقع مختلف، مثل القدم، مما سيؤدي إلى زمن أطول من أي من جهة، يرجى الاطلاع لتفاصيل 3.7.</li>
	<li>خلفية سجل أول نشاط التخطيط الدماغي. بينما يتم عقد اليد للوليد، تعليم تسجيل EEG يدوياً لتسجيل فترات حيث يتم تطبيق لا محفزات وهو يستريح الرضيع.</li>
	<li>إجراء المبلغ المطلوب من المحفزات المجس tip مسطحة-وفقا لتصميم الدراسة على اليد اليمنى للوليد. في لدينا التصميم التجريبي، قم بتعيين عدد المحفزات مسطحة-نصيحة المسبار الساعة 50. كن حذراً استخدام المسبار نصيحة مسطحة عمودياً إلى اليد للوليد حتى لا ينحني التلميح وهو تطبيق القوة الصحيح. أداء المحفزات مسبار نصيحة مسطحة باستخدام الحد أدنى من الفاصل الزمني بين التحفيز (ISI) ق 2 إلى 36 لتجنب الجمع.</li>
	<li>إيقاف كافة التسجيلات.</li>
	<li>الوثيقة تفاصيل الإعداد التجريبية.</li>
</ol>3-بيانات التحليل
<ol><li>تصفية البيانات التخطيط الدماغي الخام دون اتصال باستخدام مرشح تمرير عال في 1 هرتز ومرشح تمرير منخفض في 30 هرتز.</li>
	<li>شريحة البيانات في الحقبات من 1,500 مرض التصلب العصبي المتعدد (500 مللي قبل إلى ms 1,000 بعد ظهور التحفيز).</li>
	<li>إجراء تصحيح خط أساس للتحفيز قبل الفاصل الزمني.</li>
	<li>يدوياً رفض الحقبات EEG المحتوية على المصنوعات اليدوية، مثل التحف الحركة والضوضاء، بعد الفحص البصري. الرجاء انظر أيضا مخطط البرنامج التعليمي: https://sccn.ucsd.edu/wiki/Chapter_01:_Rejecting_Artifacts</li>
	<li>متوسط الحقبات EEG (للاستجابة الخلفية والتحفيز بشكل منفصل).</li>
	<li>ودي تصفية البيانات مع رجفان الحد أقصى من ± 50 مللي ثانية في وقت النافذة 0-1,000 ms بعد بداية التحفيز. وهذا يسمح لاختفاء الفروق بين الرضع. القيام بهذا للردود الخلفية والتحفيز بشكل منفصل.</li>
	<li>مشروع قالب نشاط الدماغ أثارت الضارة، التي تم تعريفها في dataset مستقلة وقد وصفت في11 إلى البيانات للتأكد من حجم الاستجابة الضارة أثارت داخل كل محاكمة الأفراد بالتفصيل في مكان آخر. وصف القالب الموجي مميزة لاستجابة ضارة أثارت ويعكس ضخامة كمية هذا الرد آثار ضارة داخل المحاكمة الفردية. ملاحظة: الإطار الزمني للمشروع القالب يعتمد على عمر الرضيع ووضع الحوافز. إذا تطبق المحفزات على اليد ثم الإطار الزمني لمصلحة ms 200-500 وظيفة التحفيز11. إذا طبقت المحفزات في القدم ثم الإطار الزمني لمصلحة ms 400-700 وظيفة التحفيز11. علما أنه تم التحقق من هذا القالب فقط حاليا للرضع بين من الحمل 34-43 أسبوعا. الأساليب التفصيلية المرتبطة باستخدام هذا القالب، ومناقشة للقيود، انظر هارتلي et al. عام 201711.</li>
	<li>تحقق من أن حجم نشاط الدماغ المؤذية أثارت تحسب باستخدام القالب هو أعلى بكثير عقب الحافز، مقارنة مع خلفية التخطيط الدماغي.</li>
	<li>تحليل التعبير الوجه السلوكية فيما بعد (كاميرا مرتبط بالتخطيط الدماغي تسجيل) باستخدام "نظام الترميز الوجه الوليد"2،13.</li>
</ol>

<ol>
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الكتاب ليس لها علاقة بالكشف عن.

النهج الذي قدم هنا يبين كيف الضارة أثارت الدماغ النشاط في حديثي الولادة يمكن أن يقاس بطريقة موضوعية باستخدام تسجيل EEG والمنبهات مسبار مسطحة-تلميح لتطبيق الحوافز الضارة التجريبية. يمكن استخدام هذا الأسلوب في ظروف سريرية مختلفة للكشف عن nociception، مثلاً في الأشخاص غير اللفظي مثل حديثي ال?...

الألم تجربة حسية وعاطفية غير سارة المرتبطة بتلف الأنسجة الفعلية أو المحتملة، أو وصف فيما يتعلق بمثل هذه الأضرار1. عدم القدرة على التواصل لفظياً لا ينفي إمكانية أن فرد يواجه الألم ولكن يجعلها صعبة للغاية لتقييم العلاج تخفيف الألم، على سبيل المثال في الأطفال حديثي الولادة2. وتستخدم العديد من المؤشرات السلوكية والفسيولوجية لتقييم الألم في المرضى غير اللفظي. وقد وضعت جداول مختلفة على مر السنين، والاختيار يعتمد على النوع من التحفيز والعمر الحملي والبيئة التي هي الولدان مطعم3،،من45. هذه أدوات تقييم الألم أما الاعتماد على تفسير التصنيفات أو وهم يطالبون بالمؤشرات الفيزيولوجية الثانوية.
في هذا الفيديو، ونحن نقدم وسيلة لتقييم nociception مع التسجيلات الكهربية الأمثل للاستخدام في الأطفال حديثي الولادة. ويعرف Nociception العصبية عملية ترميز المنبهات الضارة. وهكذا، كوانتيتاتينج nociception أسلوب أنيقة وموضوعية لتحديد المدخلات العصبية في شخص غير اللفظي. وعلاوة على ذلك، يرتبط بنشاط القشرية الكشف عنها بواسطة المخ (EEG) مع شدة الأحداث الضارة5،6.
الطريقة المعروضة هنا يجمع بين تسجيل EEG المنبهات الضارة التي تنتجها حفز ميكانيكية مع تحقيق نصيحة مسطحة، وتسمى أيضا وخز7، الذي ليس هو كسر الجلد ولا يسبب الكرب السلوكية6. فقد ثبت، وأن يلي nociception نقط التحفيز هو الغالب بوساطة Aδ-الألياف، هل لا حاجة جلد كسر الآفة8 وضخامة nociceptive حدة المحتملة لا يعتمد على الدولة9من النوم. التحقيق جيدا قبل أيضا الآباء والأمهات عند تطبيق هذا الأسلوب في إعداد دراسة مع الولدان. التحقيق يرتبط إلكترونيا إلى نظام تسجيل EEG تمكين تسجيل EEG تكون دقة معلم عند التحقيق بالاتصال الجلد. وهذا إلى حد كبير يبسط عملية تأمين الوقت وهو المقدمة المنطقية لجميع التحاليل اللاحقة في التخطيط الدماغي. من أجل تقليل وقت الإعداد لتسجيل EEG الرضع استخدمنا نظام تنسيب كهربائي 10/20 دولية معدلة حيث خفضنا عدد الأقطاب للحد الأدنى من المتطلبات من ثلاثة أقطاب (الشكل 1). واستخدمت مسرى تشيكوسلوفاكيا ذروة المركزية، حيث يكون نشاط الدماغ المؤذية أثارت القصوى9،،من1011، جنبا إلى جنب مع مرجع واحد وواحدة أرض القطب.

<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

الألم تجربة حسية وعاطفية غير سارة. في المرضى غير اللفظي، ومن الصعب جداً قياس الألم، حتى مع أدوات تقييم الألم. هذه الأدوات هي ذاتية أو تحديد المؤشرات الفيزيولوجية الثانوية لها أيضا بعض القيود لا سيما عند استكشاف فعالية التسكين. كما تجهيز القشرية ضروري لإدراك الألم، قد توفر تدابير نشاط الدماغ نهجاً مفيداً لتقييم الألم عند الرضع. هنا نقدم طريقة لتقييم nociception مع تسجيلات نشاط الدماغ الكهربية الأمثل للاستخدام في الأطفال حديثي الولادة. لإنتاج موحدة للغاية واستنساخه من المنبهات الضارة طبقنا التحفيز الميكانيكية مع تحقيق نصيحة مسطحة، مثلاً، المزعجة، التي ليست كسر الجلد ولا يسبب الكرب السلوكية. يسمح المؤذية أثارت إمكانية القياس الموضوعي من nociception في المرضى غير اللفظي. يمكن استخدام هذا الأسلوب في الأطفال حديثي الولادة في أقرب وقت 34 أسبوعا عمر الحمل. وعلاوة على ذلك، فإنه يمكن تطبيقها في حالات مختلفة مثل قياس فعالية العقاقير مسكن أو مخدر.

ويمثل الشكل 1 تمثيل البيانية القطب تحديد المواقع باستخدام النظام المعدل الدولي موضع القطب 10/20. ويبين الشكل 2 نشاط التخطيط الدماغي سجلت قبل فترة وجيزة، وبعد تطبيق الحوافز الضارة واحد واحد استخدام مجس نصيحة مسطحة مع 32 mN القوة، حدث التحفيز كم...

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

قياس نشاط الدماغ المؤذية أثارت في حديثي الولادة باستخدام مجس نصيحة مسطحة بالإضافة إلى المخ الكهربية

وتحد قياس الألم في المرضى غير اللفظي. في هذه الدراسة قمنا بضم تسجيل EEG مع تحفيز استخدام مجس مسطحة-تلميح للكشف عن نشاط المخ آثار ضارة بطريقة موضوعية.

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 Views.  University of Basel Children's Hospital. وتحد قياس الألم في المرضى غير اللفظي. في هذه الدراسة قمنا بضم تسجيل EEG مع تحفيز استخدام مجس مسطحة-تلميح للكشف عن نشاط المخ آثار ضارة بطريقة موضوعية.

Video: 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.

تجزئة والقياس من وحدات تخزين الدهون في الفئران نماذج السمنة عن طريق الأشعة السينية التصوير المقطعي

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

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.

فوق الجافية قياس الضغط داخل الجمجمة في الفئران باستخدام محول ضغط الألياف الضوئية

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.

قياس وعلاج الحول في قمع

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.

حالة من الفن الجمجمة التصوير بالموجات فوق الصوتية في حديثي الولادة

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.

الجزيئية التحقيق الأمثل لتحديد معدل الوفيات خلية في الكائن الضوئي (<em&gt; Microcystis الزنجارية</em&gt;) باستخدام التدفق الخلوي

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.

عبر الجلد الأوعية الدقيقة التصوير في الخدج حديثي الولادة

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.

باستخدام Saccadometry مع ديب تحفيز الدماغ لدراسة العادية والمرضية وظيفة الدماغ

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.

المعادن التصوير في أنسجة المخ عن طريق الاستئصال بالليزر - إلى جانب بالحث البلازما - الطيف الكتلي (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.

الكمي المطلق للAβ<sub&gt; 1-42</sub&gt; في السائل الدماغي الشوكي عن طريق مرجع إجراء قياس كتلة الطيف

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.

القياس المتكرر التنفسي آخر العضلات والتهوية في نماذج الماوس من مرض عصبي عضلي

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