Bölüm 1: ayarlama ve cildin hazırlanması <ol><li> Kemer, EMG, EKG, deri iletkenlik, sıcaklık, toprak nefes aşağıdaki sensörleri; </li><li> Cilt hafifçe ovuşturarak aşağıdaki konumlarda bulunan bir alkol ile silin hazırlayın: EMG gözleri yukarıda alnına, iç EKG için her iki bilek, sol elin arka zemin için (kullanılan ekipman bağlı olarak değişebilir toprak elektrot yeri); </li><li> Aşağıdaki konumlarda cilt sensörleri takın: EMG EKG her bileğinizin iç kısmında, her bir gözün üzerinde alnına, beline kemer solunum, deri, sol elinin orta parmak iletkenlik, sol elin işaretçi üzerinde termometre, zemin sol elin arkasında; </li><li> Solunum + HRV için oturum seçin; </li><li> Impendence müşterinin hareket ve makul okuma üreten duyarlı sensörler tüm sensörler için kabul edilebilir olduğunu ve emin olun. </li></ol> Bölüm 2: Kayıt <ol><li> Istemci işlem açıklayın. </li><li> (0-2 dk) açık gözlerle sessizce oturmak için istemci bilgilendirin. Bir iki dakikalık temel kaydedin. </li><li> (2-4 dk), iki dakika sonunda, bir iki dakikalık bilişsel stres başlamak aritmetik problemleri, yavaş yavaş, kalem ve kağıt veya hesap makinesi (kullanmadan çözülecek zorluk artan aritmetik müşteri için özellikle zor ise, onlara sormak defalarca yani), 100 ile &quot;seri bullak&quot; ile başlayan yedi çıkarmak için. Istemci için aşağıdaki bilgileri verin: &quot;sonraki 2 dakika içinde, ben bazı matematik problemlerini çözmek için soran olacak. Lütfen mümkün olduğu kadar çabuk çalışmak ve &quot;Bana cevabı söyler. Müşterinin cevabın doğru veya yanlış olup olmadığı geribildirim sağlamak ve böylece hafif egzersiz stres indükleyici özelliklerini ortadan kaldırmak için minimum teşvik sağlamak (örn., &quot;en iyi&quot;). </li><li> (4-6 dk) ​​istemci bu değerlendirmenin parçası olduğunu bilmek ve onu sormak / 2 dakika boyunca açık gözlerle sessizce oturmak için izin verin. </li><li> (6-8 dk) fiziksel stres başlayın: yüksek sesle zararlı gürültü açmak (bebeklerin ağlama, araba kazaları, vs) ve ayar olmadan seslerini dinlemek için istemci sormak. </li><li> (8-10 dk) gürültü kapatın ve 2 dakika süreyle açık gözlerle sessizce oturmak için istemci isteyin. </li><li> (10-12 dakika) duygusal stres başlayın: çok iyi hatırlıyorum stresli bir olay hakkında konuşmak için istemci sormak, düşünme ve hissetme ne olayın ayrıntıları yer alıyor. Stres tepkisini azaltmak için çok az empatik tepki sağlayın. </li><li> (12-14 dakika) o / o bitmiş olsa bile, istemci hikayesi kesin ve açık gözlerle sessizce oturmak için istemci talimat. </li><li> Kaydı durdurmak, verileri kaydetmek ve istemci değerlendirme yapılır bildirin. </li></ol> Bölüm 3: Bilgilendirme ve tedavi planlaması <ol><li> Deneyimleri hakkında değerlendirme sırasında istemci isteyin. Hangi parçalar özellikle stresli ve neden? O / o herhangi bir fizyolojik değişiklikler oluyor hissediyorum mı? </li><li> Ek I&#39;de tarif edilen kriterlere göre Gözden veri </li><li> Yukarıdaki analiz temelinde ele alınması gereken fizyolojik parametrelere seçti. Başlangıçta sensör okuma normalden daha yüksek veya düşük ise, tedavi hedefi istemci stresli olmayan bir ortamda normal fizyolojik parametre getirmek yardımcı olmaktır. Stres sırasında sensörü okuma normalden daha yüksek veya düşük ise, tedavi hedefi müşteri streslerinin belirlemek ve stres altında fizyolojik işleyişinin normal düzeyde tutmak için mümkün yardımcı olmaktır. Aşağı süre boyunca sensör okuma kurtarma eksikliği gösterir, tedavi amacı istemci fizyolojik parametre hızlı bir stres sonrasında normal seviyeye getirmeye yardımcı olması için. Herhangi bir veya tüm bu hedeflere tedavi planı dahil edilebilir. </li></ol> Ek I Stres değerlendirme yorumlama kurallar (Tüm &quot;cesur&quot; yanıtları alanlarının ele alınması göstermek) <table border="0"><tbody><tr><td colspan="3"> EMG (normal okuma &lt;3 mikrovolt) </td></tr><tr><td> 1. EMG başlangıçta yükselmiş midir? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. EMG 3 streslerinin herhangi bir temel geçmiş yükselmiş midir? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri boyunca temel seviyesine EMG iyileşme var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td colspan="3"> Kalp hızı (normal aralık, dakikada 60-80 atım ) </td></tr><tr><td> 1. Başlangıçta yüksek kalp hızı mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Kalp hızı, 3 streslerinin herhangi bir temel geçmiş yükselmiş midir? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri sırasında kalp atım hızı bazal seviyeye kurtarma var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td colspan="3"> Nefes alıp verme hızı (normal oranı &lt;dakikada 12 nefes) </td></tr><tr><td> 1. Başlangıçta yüksek solunum hızı mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Herhangi bir stres için temel geçmiş nefes oranı yükselmiş? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri boyunca temel oranı nefes kurtarma var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td colspan="3"> Cilt iletkenlik </td></tr><tr><td> 1. Başlangıçta deri iletkenliği yüksek midir? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Deri iletkenlik, herhangi bir stres için temel geçmiş yükselmiş var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Başlangıca deri iletkenlik kurtarma molalarda var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td colspan="3"> Parmak sıcaklık (normal okuma&gt; 90F) </td></tr><tr><td> 1. Başlangıçta düşük parmak sıcaklığı mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Parmak sıcaklığında azalma, herhangi bir stres temel geçmiş mı? </td><td> y es _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri sırasında bazal parmak ısı geri kazanımı var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td colspan="3"> Kalp hızı değişkenliğinin (normları yaşa göre değişir) </td></tr><tr><td> 1. Başlangıçtaki kalp hızı değişkenliğinin düşük mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Kalp hızı değişkenliğinin azalması için temel geçmiş herhangi bir stres mi? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri sırasında kalp hızı değişkenliğinin iyileşme var mı? </td><td> evet _ </td><td> hayır _ </td></tr></tbody></table> Ek I Stres değerlendirme yorumu: <table border="0"><tbody><tr><td colspan="3"> EMG (normal okuma &lt;3 mikrovolt) </td></tr><tr><td> 1. EMG başlangıçta yükselmiş midir? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. EMG 3 streslerinin herhangi bir temel geçmiş yükselmiş midir? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri boyunca temel seviyesine EMG iyileşme var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> Kalp hızı (normal aralık, dakikada 60-80 atım ) </td></tr><tr><td> 1. Başlangıçta yüksek kalp hızı mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Kalp hızı, 3 streslerinin herhangi bir temel geçmiş yükselmiş midir? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri sırasında kalp atım hızı bazal seviyeye kurtarma var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> Nefes alıp verme hızı (normal oranı &lt;dakikada 12 nefes) </td></tr><tr><td> 1. Başlangıçta yüksek solunum hızı mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Herhangi bir stres için temel geçmiş nefes oranı yükselmiş? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri boyunca temel oranı nefes kurtarma var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> Cilt iletkenlik (normal okuma &lt;5 mikro ohm) </td></tr><tr><td> 1. Başlangıçta deri iletkenliği yüksek midir? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Deri iletkenlik, herhangi bir stres için temel geçmiş yükselmiş var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Başlangıca deri iletkenlik kurtarma molalarda var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> Parmak sıcaklık (normal okuma&gt; 90F) </td></tr><tr><td> 1. Başlangıçta düşük parmak sıcaklığı mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Parmak sıcaklığında azalma, herhangi bir stres temel geçmiş mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 3. Dinlenme süreleri sırasında bazal parmak ısı geri kazanımı var mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> Kalp hızı değişkenliğinin (normları yaşa göre değişir) </td></tr><tr><td> 1. Başlangıçtaki kalp hızı değişkenliğinin düşük mı? </td><td> evet _ </td><td> hayır _ </td></tr><tr><td> 2. Kalp hızı değişkenliğinin azalması için temel geçmiş herhangi bir stres mi? </td><td> evet _ </td><td> hayır _ </td></tr><tr> &lt;td&gt; 3. Dinlenme süreleri sırasında kalp hızı değişkenliğinin iyileşme var mı? </td><td> evet _ </td><td> hayır _ </td></tr></tbody></table> Tedavi planlaması: Bir okuma başlangıçta normalden daha yüksek veya daha düşük ise, tedavi amacı her koşulda normal fizyolojik parametre istemci getirmeye yardımcı olması için Bir okuma stres sırasında normalden daha yüksek veya daha düşük ise, tedavi amacı istemci streslerinin belirlemek ve stres altında fizyolojik işleyişinin normal düzeyde tutmak için mümkün yardımcı olmaktır Aşağı zaman bir okuma sırasında kurtarma eksikliği gösterir, tedavinin amacı, istemci fizyolojik parametre hızlı bir stres sonrasında normal seviyeye getirmeye yardımcı olması için.

<ol>
	<li>Arena, J., Schwartz, M. S., Schwartz, M. S., Andrasik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psychophysiological+assessment+and+biofeedback+baselines.">Psychophysiological assessment and biofeedback baselines.</a> A Practitioners Guide. , 128-158 (2005).</li><li>Schwartz, N. M., Schwartz, M. S., Schwartz, M. S., Andrasik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Definitons+of+biofeedback+and+applied+psychophysiology.">Definitons of biofeedback and applied psychophysiology.</a> A Practitioners Guide. , 27-42 (2005).</li></ol>

The authors have nothing to disclose.

Appendi stres değerlendirmesinde Tamamlanan psikofizyolojik bir tedavi planı oluşturulması için gerekli bilgi ile biofeedback uygulayıcısı sağlar. Bu değerlendirme, bireyin tepkisi stereotipi, ya da tüm stres bazı fizyolojik yanıtlar birliğini ve uyaran-yanıt özgüllüğü, ya da farklı türde stresör bireyin kendine has fizyolojik yanıtlar hem de hakkında bilgi sağlar. Bu bilgi ile, istemci tür fizyolojik yanıtı hangi durumda bekliyoruz ve buna göre onun / onun cevabı değiştirmek mümkün ne...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Physiolab I-330-C2</td>
<td>&nbsp;</td>
<td>J&J</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>PLUS 12 ch</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

psychophysiological stress assessment using biofeedback 

Son yarım yüzyıl içinde, biofeedback araştırma, insan zihninin, otonom sinir sisteminin işleyişini etkileyebilir ölçüde göstermiştir, önceden bilinçli kontrolü dışında olduğu düşünülmektedir. Insanlar kendi bedenleri sinyalleri gözlemlemek izin vererek, biofeedback, stres gibi fizyolojik ve psikolojik reaksiyonlar, daha fazla farkındalık geliştirmek için sağlar, ve bu reaksiyonlar değiştirmek öğrenmek için. Biofeedback uygulayıcıları insanların reaksiyonları hafif stresli olayların değerlendirilmesi ve biofeedback tabanlı bir tedavi planı formüle bu süreci kolaylaştırabilir. Stres değerlendirilmesi sırasında ilk uygulayıcısı fizyolojik okumaları için bir temel kayıtları, ve sonra, bilişsel, fiziksel ve duygusal stres gibi çeşitli hafif stres ile istemci sunuyor. Stresör Variety bir kişinin uyaran-yanıt özgüllük, ya da her kişinin niteliksel olarak farklı uyaranlara tepki olarak farklılıkları belirlemek amacıyla sunulmaktadır. Bu video, tedavi planlaması için biofeedback ve şimdiki genel kurallar kullanarak psikofizyolojik stres değerlendirme süreci gözler önüne serecektir.

Watch this Scientific Journal Video about Psychophysiological Stress Assessment Using Biofeedback at JoVE.com

Psychophysiological Stress Assessment Using Biofeedback

Bu video, biofeedback kullanarak stres kişinin psikofizyolojik reaksiyon değerlendirilmesinde bir yöntem anlatacağım. Ayrıca, tedavi planlaması için genel kurallar sunacak.

Biofeedback Kullanımı Psikofizyolojik Stres Değerlendirme

In the last half century, research in biofeedback has shown the extent to which the human mind can influence the functioning of the autonomic nervous ...

psychophysiological-stress-assessment-using-biofeedback

Research

JoVE Journal

Biology

Psychophysiological Stress Assessment Using Biofeedback 

13.2K Görüntüleme.  Cambridge Health Alliance. Bu video, biofeedback kullanarak stres kişinin psikofizyolojik reaksiyon değerlendirilmesinde bir yöntem anlatacağım. Ayrıca, tedavi planlaması için genel kurallar sunacak.

Video: Psychophysiological Stress Assessment Using Biofeedback

Monitoring Plant Hormones During Stress Responses

Plant hormones and related signaling compounds play an important role in the regulation of plant responses to various environmental stimuli and stresses. Among the most severe stresses are insect herbivory, pathogen infection, and drought stress. For each of these stresses a specific set of hormones and/or combinations thereof are known to fine-tune the responses, thereby ensuring the plant's survival. The major hormones involved in the regulation of these responses are jasmonic acid (JA), salicylic acid (SA), and abscisic acid (ABA). To better understand the role of individual hormones as well as their potential interaction during these responses it is necessary to monitor changes in their abundance in a temporal as well as in a spatial fashion. For the easy, sensitive, and reproducible quantification of these and other signaling compounds we developed a method based on vapor phase extraction and gas chromatography/mass spectrometry (GC/MS) analysis (1, 2, 3, 4). After extracting these compounds from the plant tissue by acidic aqueous 1-propanol mixed with dichloromethane the carboxylic acid-containing compounds are methylated, volatilized under heat, and collected on a polymeric absorbent. After elution into a sample vial the analytes are separated by gas chromatography and detected by chemical ionization mass spectrometry. The use of appropriate internal standards then allows for the simple quantification by relating the peak areas of analyte and internal standard.

A simple method is provided that allows for the rapid extraction and analysis of multiple plant hormones from small tissue samples. The procedure uses vapor phase extraction as the solemn purification step. Samples are analyzed by GC/MS with chemical ionization that produces mainly (M+1)+ ions.

Stres Cevaplarının sırasında Bitki Hormonları İzlenmesi

Plant hormones and related signaling compounds play an important role in the regulation of plant responses to various environmental stimuli and stresses. ...

Biyoloji

Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important treatment for aneurysms which attempts to exclude blood from flowing into the aneurysmal segment of the vessel while preserving blood flow in a normal fashion.  Improper clip placement may result in residual aneurysm with the potential for subsequent aneurysm rupture or partial or full occlusion of distal arteries resulting in cerebral infarction.  Here we describe the use of an ultrasonic flow probe to provide quantitative evaluation of arterial flow before and after microsurgical clip placement at the base of a middle cerebral artery aneurysm.  This information helps ensure adequate aneurysm reconstruction with preservation of normal distal blood flow.

Description of the surgical obliteration of a cerebral aneurysm utilizing an ultrasonic flow probe to assess arterial flow prior to and after aneurysm clip placement.

İntraoperatif Akış Değerlendirme Orta Serebral Anevrizma Mikrocerrahi Klip obliterasyonu

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important ...

A Quantitative Assessment of The Yeast Lipidome using Electrospray Ionization Mass Spectrometry

Lipids are one of the major classes of biomolecules and play important roles membrane dynamics, energy storage, and signalling1-4. The budding yeast Saccharomyces cerevisiae, a genetically and biochemically manipulable unicellular eukaryote with annotated genome and very simple lipidome, is a valuable model for studying biological functions of various lipid species in multicellular eukaryotes2,3,5. S. cerevisiae has 10 major classes of lipids with chain lengths mainly of 16 or 18 carbon atoms and either zero or one degree of unsaturation6,7. Existing methods for lipid identification and quantification - such as high performance liquid chromatography, thin-layer chromatography, fluorescence microscopy, and gas chromatography followed by MS - are well established but have low sensitivity, insufficiently separate various molecular forms of lipids, require lipid derivitization prior to analysis, or can be quite time consuming. Here we present a detailed description of our experimental approach to solve these inherent limitations by using survey-scan ESI/MS for the identification and quantification of the entire complement of lipids in yeast cells. The described method does not require chromatographic separation of complex lipid mixtures recovered from yeast cells, thereby greatly accelerating the process of data acquisition. This method enables lipid identification and quantification at the concentrations as low as g/ml and has been successfully applied to assessing lipidomes of whole yeast cells and their purified organelles. Lipids extraction from whole yeast cells for using this method of lipid analysis takes two to three hours. It takes only five to ten minutes to run each sample of extracted and dried lipids on a Q-TOF mass spectrometer equipped with a nano-electrospray source.

We describe a new quantitative lipidomics method for identifying numerous lipid species in yeast using survey-scan electrospray ionization mass spectrometry (ESI/MS). This method exceeds currently available methods for lipid identification and quantification in the ability to resolve various molecular forms of lipids, sensitivity, and speed.

Nicel Değerlendirme Elektrosprey İyonizasyon Kütle Spektrometresi kullanarak Maya Lipidome

Lipids are one of the major classes of biomolecules and play important roles membrane dynamics, energy storage, and signalling1-4. The budding yeast ...

The use of Biofeedback in Clinical Virtual Reality: The INTREPID Project

Generalized anxiety disorder (GAD) is a psychiatric disorder characterized by a constant and unspecific anxiety that interferes with daily-life activities. Its high prevalence in general population and the severe limitations it causes, point out the necessity to find new efficient strategies to treat it. Together with the cognitive-behavioral treatments, relaxation represents a useful approach for the treatment of GAD, but it has the limitation that it is hard to be learned. The INTREPID project is aimed to implement a new instrument to treat anxiety-related disorders and to test its clinical efficacy in reducing anxiety-related symptoms. The innovation of this approach is the combination of virtual reality and biofeedback, so that the first one is directly modified by the output of the second one. In this way, the patient is made aware of his or her reactions through the modification of some features of the VR environment in real time. Using mental exercises the patient learns to control these physiological parameters and using the feedback provided by the virtual environment is able to gauge his or her success. The supplemental use of portable devices, such as PDA or smart-phones, allows the patient to perform at home, individually and autonomously, the same exercises experienced in therapist's office. The goal is to anchor the learned protocol in a real life context, so enhancing the patients' ability to deal with their symptoms. The expected result is a better and faster learning of relaxation techniques, and thus an increased effectiveness of the treatment if compared with traditional clinical protocols.

The Project on Virtual Reality Intelligent Multi-sensor System for the Treatment of Anxiety-related Disorders (INTREPID) is aimed at developing a multi-sensor context-aware virtual reality system for the treatment of anxiety-related disorders.

Klinik Sanal Gerçeklik Biofeedback kullanımı: INTREPID Projesi

Generalized anxiety disorder (GAD) is a psychiatric disorder characterized by a constant and unspecific anxiety that interferes with daily-life ...

Assessment of GFP Expression and Viability Using the Tali Image-Based Cytometer

Single-cell and population information are commonly obtained either by flow cytometry or fluorescence microscopy. However, these two methods provide different information. Flow cytometry gives quantitative multi-parametric information about physical characteristics and staining or expression, but doesn't allow for visualization. Stand-alone fluorescence microscopy provides visual data, but doesn't allow for straightforward quantitative measurements1.
Image-based cytometry bridges the gap between these two methods, enabling the quick visualization and simultaneous quantitative analysis of thousands of cells in heterogeneous populations2. Here, we present a method for performing cell viability and green fluorescent protein (GFP) expression assays using the Tali Image-Based Cytometer3. The Tali instrument is a 3-channel (bright field, green fluorescence, red fluorescence) benchtop assay platform that offers several advantages over flow cytometry and fluorescence microscopy. The Tali cytometer is less expensive, takes up less bench space, requires less maintenance, and the work flow has been simplified so that the operation and analysis is much simpler and quicker. The Tali cytometer is capable of performing a range of suspension cell-based assays, including GFP and red fluorescent protein (RFP) expression, apoptosis4-6 and cell viability analysis with propidium iodide (PI)7-11.
Here, we demonstrate the use of the Tali instrument in performing a cell viability assay in cells expressing GFP. GFP-transduced cells are stained using the Tali Viability Kit - Dead Cell Red. The cells are then pipetted into a Tali Cellular Analysis Slide and loaded into the cytometer. Bright field, red fluorescence and green fluorescence images are captured and analyzed using assay specific algorithms. Histograms are then generated to display cell size, PI fluorescence intensity, and GFP fluorescence intensity. These parameters can then be thresholded to home in on a specific cell population.
A side-by side comparison of the Tali Image-Based Cytometer and traditional flow cytometry demonstrates that the two methods provide comparable data regarding cell viability and protein expression. However, the Tali instrument provides additional visual information about the cell population that cannot be obtained using a flow cytometer.

This protocol describes how to perform cell viability and fluorescence expression assays using the Tali Image-Based Cytometer.

Tali Görüntü Tabanlı Sitometre kullanarak GFP İfade ve Canlılık Değerlendirilmesi

Single-cell and population information are commonly obtained either by flow cytometry or fluorescence microscopy. However, these two methods provide ...

Analysis of Oxidative Stress in Zebrafish Embryos

High levels of reactive oxygen species (ROS) may cause a change of cellular redox state towards oxidative stress condition. This situation causes oxidation of molecules (lipid, DNA, protein) and leads to cell death. Oxidative stress also impacts the progression of several pathological conditions such as diabetes, retinopathies, neurodegeneration, and cancer. Thus, it is important to define tools to investigate oxidative stress conditions not only at the level of single cells but also in the context of whole organisms. Here, we consider the zebrafish embryo as a useful in vivo system to perform such studies and present a protocol to measure in vivo oxidative stress. Taking advantage of fluorescent ROS probes and zebrafish transgenic fluorescent lines, we develop two different methods to measure oxidative stress in vivo: i) a &#8220;whole embryo ROS-detection method&#8221; for qualitative measurement of oxidative stress and ii) a &#8220;single-cell&#160;ROS detection method&#8221; for quantitative measurements of oxidative stress. Herein, we demonstrate the efficacy of these procedures by increasing oxidative stress in tissues by oxidant agents and physiological or genetic methods. This protocol is amenable for forward genetic screens and it will help address cause-effect relationships of ROS in animal models of oxidative stress-related pathologies such as neurological disorders and cancer.

Here we report a protocol to measure oxidative stress in living zebrafish embryos. This procedure allows reactive oxygen species (ROS) detection in both whole embryo tissues and single-cell populations. This protocol will accomplish both qualitative and quantitative analyses.

Zebra balığı Embriyolar Oksidatif Stres Analizi

High levels of reactive oxygen species (ROS) may cause a change of cellular redox state towards oxidative stress condition. This situation causes ...

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of central nervous system injuries in vivo, possibly by reducing oxidative stress. However, effects of R/NIR-LT on oxidative stress have been shown to vary depending on wavelength or intensity of irradiation. Studies comparing treatment parameters are lacking, due to absence of commercially available devices that deliver multiple wavelengths or intensities, suitable for high through-put in vitro optimization studies. This protocol describes a technique for delivery of light at a range of wavelengths and intensities to optimize therapeutic doses required for a given injury model. We hypothesized that a method of delivering light, in which wavelength and intensity parameters could easily be altered, could facilitate determination of an optimal dose of R/NIR-LT for reducing reactive oxygen species (ROS) in vitro.
Non-coherent Xenon light was filtered through narrow-band interference filters to deliver varying wavelengths (center wavelengths of 440, 550, 670 and 810nm) and fluences (8.5 x 10-3 to 3.8 x 10-1 J/cm2) of light to cultured cells. Light output from the apparatus was calibrated to emit therapeutically relevant, equal quantal doses of light at each wavelength. Reactive species were detected in glutamate stressed cells treated with the light, using DCFH-DA and H2O2 sensitive fluorescent dyes.&#160;
We successfully delivered light at a range of physiologically and therapeutically relevant wavelengths and intensities, to cultured cells exposed to glutamate as a model of CNS injury. While the fluences of R/NIR-LT used in the current study did not exert an effect on ROS generated by the cultured cells, the method of light delivery is applicable to other systems including isolated mitochondria or more physiologically relevant organotypic slice culture models, and could be used to assess effects on a range of outcome measures of oxidative metabolism.

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

Non-coherent Xenon light was passed through narrow-band interference and neutral density filters to deliver light of varying wavelength and intensity to cultured cells. This protocol was used to assess the effects of red/near-infrared light therapy on production of reactive species in vitro: no effects were observed using the tested parameters.

Dalga boyundaki ve Oksidatif Stres Kırmızı / yakın-kızılötesi ışık Terapisi şiddetler bir Range Etkilerinin Değerlendirilmesi için Yöntemi<em&gt; İn Vitro</em

Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of ...

A Protocol for Using Gene Set Enrichment Analysis to Identify the Appropriate Animal Model for Translational Research

Recent studies that compared transcriptomic datasets of human diseases with datasets from mouse models using traditional gene-to-gene comparison techniques resulted in contradictory conclusions regarding the relevance of animal models for translational research. A major reason for the discrepancies between different gene expression analyses is the arbitrary filtering of differentially expressed genes. Furthermore, the comparison of single genes between different species and platforms often is limited by technical variance, leading to misinterpretation of the con/discordance between data from human and animal models. Thus, standardized approaches for systematic data analysis are needed. To overcome subjective gene filtering and ineffective gene-to-gene comparisons, we recently demonstrated that gene set enrichment analysis (GSEA) has the potential to avoid these problems. Therefore, we developed a standardized protocol for the use of GSEA to distinguish between appropriate and inappropriate animal models for translational research. This protocol is not suitable to predict how to design new model systems a-priori, as it requires existing experimental omics data. However, the protocol describes how to interpret existing data in a standardized manner in order to select the most suitable animal model, thus avoiding unnecessary animal experiments and misleading translational studies.

We provide a standardized protocol for the use of gene set enrichment analysis of transcriptomic data to identify an ideal mouse model for translational research. 
This protocol can be used with DNA microarray and RNA sequencing data and can further be extended to other omics data if data are available.

Gen kullanmak için bir protokol translasyonel araştırma için uygun hayvan modeli belirlemek için zenginleştirme çözümlemesi ayarla

Recent studies that compared transcriptomic datasets of human diseases with datasets from mouse models using traditional gene-to-gene comparison ...

Fish Sperm Assessment Using Software and Cooling Devices

For gamete quality evaluation, there are innovative, rapid, and quantitative techniques that can provide useful data for aquaculture. Computerized systems for sperm analysis were developed to measure several parameters and one of the most commonly measured is the sperm motility.
Initially, this computer technology was designed for mammalian species, although it can also be used for fish sperm analysis. Fish have specific features that can affect sperm assessment such as a short motility time after activation and, in some cases, adaptation to lower temperatures. Thus, it is necessary to modify both software and hardware components to make motility analysis more efficient for fish sperm analysis. For mammalian sperm, the heating plate is used to maintain optimal temperatures of spermatozoa. However, for some fish species, it is advantageous to use a lower temperature to prolong the duration of motility, since the sperm remain active for less than 2 min. Therefore, cooling devices are necessary to refrigerate samples at constant temperature over the time of analysis, including on the optical microscope. This protocol describes the analysis of fish sperm motility using software for sperm analysis and new cooling devices to optimize the results.

The present protocol describes a procedure of fish sperm assessment using computer-assisted sperm analysis and cooling devices. The software gives a rapid, accurate and quantitative analysis of fish sperm quality based on spermatozoa motility, which can be a useful tool in aquaculture to improve reproduction success.

Yazılımı kullanarak ve soğutma aygıtlarının balık Sperm değerlendirmesi

For gamete quality evaluation, there are innovative, rapid, and quantitative techniques that can provide useful data for aquaculture. Computerized systems ...

An Air-liquid Interface Bronchial Epithelial Model for Realistic, Repeated Inhalation Exposure to Airborne Particles for Toxicity Testing

For toxicity testing of airborne particles, air-liquid interface (ALI) exposure systems have been developed for in vitro tests in order to mimic realistic exposure conditions. This puts specific demands on the cell culture models. Many cell types are negatively affected by exposure to air (e.g., drying out) and only remain viable for a few days. This limits the exposure conditions that can be used in these models: usually relatively high concentrations are applied as a cloud (i.e., droplets containing particles, which settle down rapidly) within a short period of time. Such experimental conditions do not reflect realistic long-term exposure to low concentrations of particles. To overcome these limitations the use of a human bronchial epithelial cell line, Calu-3 was investigated. These cells can be cultured at ALI conditions for several weeks while retaining a healthy morphology and a stable monolayer with tight junctions. In addition, this bronchial model is suitable for testing the effects of repeated exposures to low, realistic concentrations of airborne particles using an ALI exposure system. This system uses a continuous airflow in contrast to other ALI exposure systems that use a single nebulization producing a cloud. Therefore, the continuous flow system is suitable for repeated and prolonged exposure to airborne particles while continuously monitoring the particle characteristics, exposure concentration, and delivered dose. Taken together, this bronchial model, in combination with the continuous flow exposure system, is able to mimic realistic, repeated inhalation exposure conditions that can be used for toxicity testing.

Described is the cell culture and exposure method of an in vitro bronchial model for realistic, repeated inhalation exposure to particles for toxicity testing.

Toksisite Testi için Havadaki Parçacıklara Gerçekçi, Tekrarlanan Soluma Maruziyeti için Hava Sıvısı Arayüzü Bronşiyal Epitel Modeli

For toxicity testing of airborne particles, air-liquid interface (ALI) exposure systems have been developed for in vitro tests in order to mimic realistic ...