作者要感谢Andrea Thomson，Jon Hash，Hope Woods和Autumn Anthony为QST处理狗，Masataka Enomoto帮助筛选狗，以及Sam Chiu为建立热热QST协议所做的贡献。

所有程序均由北卡罗来纳州立大学机构动物护理和使用委员会批准。
1. 房间设置和学习科目适应
<ol><li>在专用空间中进行 QST，该空间有足够的空间供 QST 操作员、处理员和任何大小的狗舒适地移动。尽量减少潜在的听觉和视觉干扰，并使用白噪声机器阻挡环境声音。</li>
	<li>在地板上放置一个大的瑜伽垫或类似的衬垫，以确保狗在测试期间在侧卧时感到舒适。</li>
	<li>让狗至少 10 分钟自由探索和适应房间，并与 QST 操作员和处理者相处融洽。在客房内随意提供淡水，并偶尔提供食物奖励。</li>
	<li>通过抛硬币随机化测试站点（左侧或右侧）。在第三和第四跖骨背表面之间的空间周围夹一个大约 2 x 4 cm 的皮毛部分，中间位于跗跖关节和跖趾关节之间。在腕骨上方靠近臂前腕关节的外侧臂前夹一个大约 1 x 2 cm 的毛皮切片。</li>
</ol>2. 电子冯弗雷麻醉仪
<ol><li>仪器设置<ol><li>轻轻地将刚性 0.9 mm 冯弗雷尖端施加到称重传感器上，并确保称重传感器牢固地拧入手机中。通过M0通道将电源线从手机连接到录音设备（图1A，B）。</li>
		<li>打开记录设备并按下 MAX 按钮，以便设备将记录并显示狗对施加的刺激做出反应时达到的最大力（图1C）。</li>
		<li>按下 CLR 按钮将仪器归零。</li>
	</ol></li>
	<li>数据采集<ol><li>将狗放在侧面卧位以测量阈值。 注意:将狗放置在右侧卧位以测量左肢的阈值，或放置在左侧卧位以测量右肢的阈值。如果狗在给予口头提示时不愿意躺在侧卧中，QST 操作员和处理员可以手动将狗放在侧卧位。</li>
		<li>根据需要应用最小到中度的约束，以保持狗在侧卧和相对静止。 注意:处理程序执行此步骤。</li>
		<li>一旦狗平静和放松并且被测试的肢体至少伸展 70%，就应用刺激。为被测试的肢体提供温和的手动支撑，使肢体远离地板，并提供稳定的支撑以施加力，同时不阻止狗抽出肢体。 注意:QST 运算符执行此步骤。</li>
		<li>垂直于被测区域的皮肤应用冯弗雷尖端。如果狗表现出反射运动（例如，爪子抽搐或在用力前肢体退出），因为冯弗雷尖端在皮肤上的感觉，让狗在重新应用冯弗雷尖端之前再次放松肢体。当皮肤不引起反射运动时，通过应用冯弗雷尖端进行测量。</li>
		<li>用冯弗雷尖端（~20 g/sec）施加稳定增加的力，直到狗撤回肢体、发声、转身看刺激，或表现出其他表明对刺激的有意识感知的动作或行为反应。当狗抽出肢体或达到最大力时，移除刺激。 注意:不要超过 1，000 克的力。</li>
		<li>记录记录设备上显示的最大力。 注意:如果达到 1，000 g 力的安全截止值，则记录 1，000 g 作为感官阈值，并且注意在安全截止之前没有响应。</li>
		<li>重复测量总共五次试验，每次测量之间留出1分钟（试验间隔）。通过按 CLR 按钮在每个步骤之间将仪器归零。<ol><li>允许狗在试验间隔期间保持侧卧，如果它们保持相对平静和放松，没有或很少的约束。否则，让狗在 QST 房间内坐下、站立或移动以保持舒适度。在随后的测量之前，将狗放回侧面卧位。</li>
		</ol></li>
		<li>记录 0-5 的可行性分数，以指示收集数据的难易程度。 注意:可行性分数如下:0 = 没有问题，1 = 轻度难度，2 = 中等难度，3 = 显着难度，4 = 极端困难，5 = 不可能。用于分配可行性分数的量规如 表1所示。</li>
		<li>让狗休息 5 分钟，然后开始用钝探头的压力测藻力计进行测量。</li>
	</ol></li>
</ol>3. 钝探头压力测数计
<ol><li>仪器设置<ol><li>确保将小钝探头牢固地拧入设备（图2A）。</li>
		<li>打开录音设备，并在屏幕上出现提示时按 MAX 按钮继续。按 UNIT 按钮，直到设备在屏幕顶部显示为克 （g）（图 2B）。</li>
		<li>按 ZERO 按钮将仪器归零。</li>
	</ol></li>
	<li>数据采集<ol><li>将狗放在侧面卧位以测量阈值。 注意:将狗放置在右侧卧位以测量左肢的阈值，或放置在左侧卧位以测量右肢的阈值。如果狗在给予口头提示时不愿意躺在侧卧中，QST 操作员和处理员可以手动将狗放在侧卧位。</li>
		<li>根据需要应用最小到中度的约束，以保持狗在侧卧和相对静止。 注意:处理程序执行此步骤。</li>
		<li>一旦狗平静和放松并且被测试的肢体伸展约 70%，就应用刺激。为被测试的肢体提供温和的手动支撑，使肢体远离地板，并提供稳定的支撑以施加力，同时不阻止狗抽出肢体。 注意:QST 运算符执行此步骤。</li>
		<li>垂直于被测区域的皮肤应用钝探针（图2C）。如果狗在皮肤上感受到钝探针后表现出反射运动（例如，爪子抽搐或在用力前肢体撤回），请在重新应用钝探针之前让狗再次放松肢体。当将钝探头施加到皮肤上不引起反射运动时进行测量。</li>
		<li>用探针施加稳定增加的力（~20 g / s），直到狗撤回肢体，发声，转身看刺激，或表现出其他表明对刺激的有意识感知的运动或行为反应。当狗抽出肢体或达到最大力时，移除刺激。 注意:不要超过 2，500 克的力。</li>
		<li>记录记录设备上显示的最大力。 注意:如果达到 2，500 g 力的安全截止值，则记录 2，500 g 作为感官阈值，并注意在安全截止之前没有响应。</li>
		<li>重复测量总共五次试验，每次测量之间留出1分钟（试验间隔）。按 ZERO 按钮，在每一步之间将仪器归零。<ol><li>允许狗在试验间隔期间保持横向卧位，如果它们保持相对平静和放松，没有或很少的约束。否则，让狗在 QST 房间内坐下、站立或移动以保持舒适度。在随后的测量之前，将狗放回侧面卧位。</li>
		</ol></li>
		<li>记录 0-5 的可行性分数，以指示收集数据的难易程度。 注意:可行性分数如下:0 = 没有问题，1 = 轻度难度，2 = 中等难度，3 = 显着难度，4 = 极端困难，5 = 不可能。</li>
		<li>让狗休息 5 分钟，然后开始使用热探头进行测量。</li>
	</ol></li>
</ol>4. 热探头
<ol><li>仪器设置<ol><li>通过USB电缆 将 热感分析仪连接到计算机，并确保16 x 16 mm温度计连接到分析仪。打开分析器。</li>
		<li>打开计算机上的热感分析仪软件，然后从启动菜单中选择TSA II分析仪。在分析器自检的弹出警告上单击 确定 。确保在自检期间，恒温器未连接到研究对象。</li>
		<li>在" 测试 "选项卡（右上角）的"选择患者"提示（屏幕左侧）下，双击列表中的姓名来 选择相应的患者 。<ol><li>要创建新患者，请单击"测试"选项卡右侧的"患者"选项卡。单击左下角的新患者图标并填写患者详细信息（部门、患者姓名、ID、性别和出生日期）。</li>
		</ol></li>
		<li>在"测试"选项卡的"选择程序"提示下，双击列表中的程序来选择相应的程序。<ol><li>要创建新程序，请单击"患者"选项卡右侧的"程序"选项卡。单击左下角的"新建程序"图标并填写程序详细信息。 注意:对于此协议，程序详细信息在表 2 中给出。不需要在"测试"选项卡的"选择正文站点"提示下选择正文站点。</li>
		</ol></li>
		<li>选择适当的患者和程序后，单击"测试"选项卡下的"转到测试"提示。单击左下角的"预测试"按钮，将分析仪校准到指定的程序。</li>
		<li>预测试完成后，"预测试"按钮将替换为"开始"按钮，并显示测试窗口:按"开始"按钮开始测试（图 3A）。</li>
		<li>松开热度仪电缆，确保热度仪易于取用。</li>
	</ol></li>
	<li>数据采集<ol><li>将狗放在侧面卧位以测量热潜伏期。 注意:将狗放置在右侧卧位以测量左肢的阈值，或放置在左侧卧位以测量右肢的阈值。如果狗在给予口头提示时不愿意躺在侧卧中，QST 操作员和处理员可以手动将狗放在侧卧位。</li>
		<li>根据需要应用最小到中度的约束，以保持狗在侧卧和相对静止。 注意:处理程序执行此步骤。</li>
		<li>一旦狗平静和放松并且被测试的肢体伸展约 70%，就应用刺激。为被测试的肢体提供温和的手动支撑，使肢体远离地板，同时不阻止狗抽出肢体。此外，用手支撑肢体握住并操作秒表。 注意:QST 运算符执行此步骤。</li>
		<li>将热化物涂在被测区域的皮肤上（图3B）。如果狗表现出反射运动（例如，在加热之前爪子抽搐或肢体退出），因为皮肤上的热德感觉，在重新涂抹热化物之前让狗再次放松肢体。当将热敷物应用于皮肤不引起反射运动时进行测量。</li>
		<li>单击"测试"选项卡左下角的"开始"按钮以开始测试。 注意:QST 操作员向处理程序发出信号以开始测试（例如，通过点头），QST 操作员同时启动秒表。</li>
		<li>当狗抽出肢体、发声、转身看刺激或表现出其他表明对刺激的有意识感知的运动或行为反应时，或者在停止秒表的同时达到最大潜伏期时，取下热物。 注意:QST 运算符执行此步骤。不要超过20秒的热化物应用或49°C的最高温度。</li>
		<li>记录提款延迟。如果达到20 s的热化物应用安全截止，记录20 s作为感官潜伏期，并注意在安全截止之前没有响应。</li>
		<li>重复测量总共五次试验，每次测量之间留出1分钟（试验间隔）。单击" 结束 测试"按钮，然后单击每次测量之间的 "预测试 "按钮，停止加热温度计并重新校准温度计，为下一次应用做准备。 注意:处理程序执行此步骤。<ol><li>允许狗在试验间隔期间保持横向卧位，如果它们保持相对平静和放松，没有或很少的约束。否则，让狗在 QST 房间内坐下、站立或移动以保持舒适度。在随后的测量之前，将狗放回侧面卧位。</li>
		</ol></li>
		<li>记录 0-5 的可行性分数，以指示收集数据的难易程度。 注意:可行性分数如下:0 = 没有问题，1 = 轻度难度，2 = 中等难度，3 = 显着难度，4 = 极端困难，5 = 不可能。</li>
	</ol></li>
</ol>

<ol>
	<li>Hunt, J., Knazovicky, D., Lascelles, B. D. X., Murrell, J. <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+dogs+with+painful+disease:+A+window+to+pain+mechanisms.">Quantitative sensory testing in dogs with painful disease: A window to pain mechanisms.</a> The Veterinary Journal. 243, 33-41 (2019).</li><li>Purves, D., 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=Cutaneous+and+subcutaneous+somatic+sensory+receptors.">Cutaneous and subcutaneous somatic sensory receptors.</a> Neuroscience, 2nd edition. , (2001).</li><li>Gorney, A. 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=Mechanical+and+thermal+sensory+testing+in+normal+chondrodystrophic+dogs+and+dogs+with+spinal+cord+injury+caused+by+thoracolumbar+intervertebral+disc+herniations.">Mechanical and thermal sensory testing in normal chondrodystrophic dogs and dogs with spinal cord injury caused by thoracolumbar intervertebral disc herniations.</a> Journal of Veterinary Internal Medicine. 30 (2), 627-635 (2016).</li><li>Song, R. B., 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=von+Frey+anesthesiometry+to+assess+sensory+impairment+after+acute+spinal+cord+injury+caused+by+thoracolumbar+intervertebral+disc+extrusion+in+dogs.">von Frey anesthesiometry to assess sensory impairment after acute spinal cord injury caused by thoracolumbar intervertebral disc extrusion in dogs.</a> The Veterinary Journal. 209, 144-149 (2016).</li><li>Moore, S. A., Hettlich, B. F., Waln, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+an+electronic+von+Frey+device+for+evaluation+of+sensory+threshold+in+neurologically+normal+dogs+and+those+with+acute+spinal+cord+injury.">The use of an electronic von Frey device for evaluation of sensory threshold in neurologically normal dogs and those with acute spinal cord injury.</a> The Veterinary Journal. 197 (2), 216-219 (2013).</li><li>Sanchis-Mora, S., 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=Pregabalin+for+the+treatment+of+syringomyelia-associated+neuropathic+pain+in+dogs:+A+randomized,+placebo-controlled,+double-masked+clinical+trial.">Pregabalin for the treatment of syringomyelia-associated neuropathic pain in dogs: A randomized, placebo-controlled, double-masked clinical trial.</a> The Veterinary Journal. 250, 55-62 (2019).</li><li>Brydges, N. 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=Clinical+assessments+of+increased+sensory+sensitivity+in+dogs+with+cranial+cruciate+ligament+rupture.">Clinical assessments of increased sensory sensitivity in dogs with cranial cruciate ligament rupture.</a> The Veterinary Journal. 193 (2), 546-550 (2012).</li><li>Williams, M. D., 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=Feasibility+and+repeatability+of+thermal+quantitative+sensory+testing+in+normal+dogs+and+dogs+with+hind+limb+osteoarthritis-associated+pain.">Feasibility and repeatability of thermal quantitative sensory testing in normal dogs and dogs with hind limb osteoarthritis-associated pain.</a> The Veterinary Journal. 199, 63-67 (2014).</li><li>Freire, M., Knazovicky, D., Case, B., Thomson, A., Lascelles, B. D. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+thermal+and+mechanical+quantitative+sensory+testing+in+client-owned+dogs+with+chronic+naturally+occurring+pain+and+normal+dogs.">Comparison of thermal and mechanical quantitative sensory testing in client-owned dogs with chronic naturally occurring pain and normal dogs.</a> The Veterinary Journal. 210, 95-97 (2016).</li><li>Knazovicky, D., 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=Widespread+somatosensory+sensitivity+in+naturally+occurring+canine+model+of+osteoarthritis.">Widespread somatosensory sensitivity in naturally occurring canine model of osteoarthritis.</a> Pain. 157 (6), 1325-1332 (2016).</li><li>Lascelles, B. D. X., Cripps, P. J., Jones, A., Waterman, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-operative+central+hypersensitivity+and+pain:+The+pre-emptive+value+of+pethidine+for+ovariohysterectomy.">Post-operative central hypersensitivity and pain: The pre-emptive value of pethidine for ovariohysterectomy.</a> Pain. 73 (3), 461-471 (1997).</li><li>Slingsby, L. S., Waterman-Pearson, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+post-operative+analgesic+effects+of+ketamine+after+canine+ovariohysterectomy+-+a+comparison+between+pre-+or+post-operative+administration.">The post-operative analgesic effects of ketamine after canine ovariohysterectomy - a comparison between pre- or post-operative administration.</a> Research in Veterinary Medicine. 69 (2), 147-152 (2000).</li><li>Sammarco, J. L., 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=Post-operative+analgesia+for+stifle+surgery:+A+comparison+of+intra-articular+bupivacaine,+morphine,+or+saline.">Post-operative analgesia for stifle surgery: A comparison of intra-articular bupivacaine, morphine, or saline.</a> Veterinary Surgery. 25 (1), 59-69 (1996).</li><li>Tomas, A., Marcellin-Little, D. J., Roe, S. C., Motsinger-Reif, A., Lascelles, B. D. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+mechanical+thresholds+and+limb+use+in+dogs+with+coxofemoral+joint+OA-associated+pain+and+the+modulating+effects+of+pain+alleviation+from+total+hip+replacement+on+mechanical+thresholds.">Relationship between mechanical thresholds and limb use in dogs with coxofemoral joint OA-associated pain and the modulating effects of pain alleviation from total hip replacement on mechanical thresholds.</a> Veterinary Surgery. 43 (5), 542-548 (2014).</li><li>Briley, J. D., Williams, M. D., Freire, M., Griffith, E. H., Lascelles, B. D. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feasibility+and+repeatability+of+cold+and+mechanical+quantitative+sensory+testing+in+normal+dogs.">Feasibility and repeatability of cold and mechanical quantitative sensory testing in normal dogs.</a> The Veterinary Journal. 199 (2), 246-250 (2014).</li><li>Knazovicky, D., 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=Replicate+effects+and+test-retest+reliability+of+quantitative+sensory+threshold+testing+in+dogs+with+and+without+chronic+pain.">Replicate effects and test-retest reliability of quantitative sensory threshold testing in dogs with and without chronic pain.</a> Veterinary Anesthesia and Analgesia. 44 (3), 615-624 (2017).</li><li>Sanchis-Mora, S., 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=Development+and+initial+validation+of+a+sensory+threshold+examination+protocol+(STEP)+for+phenotyping+canine+pain+syndromes.">Development and initial validation of a sensory threshold examination protocol (STEP) for phenotyping canine pain syndromes.</a> Veterinary Anesthesia and Analgesia. 44 (3), 600-614 (2017).</li><li>Backonja, 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=Value+of+quantitative+sensory+testing+in+neurological+and+pain+disorders:+NeuPSIG+consensus.">Value of quantitative sensory testing in neurological and pain disorders: NeuPSIG consensus.</a> Pain. 154 (9), 1807-1819 (2013).</li><li>Wylde, V., Palmer, S., Learmonth, I. D., Dieppe, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Somatosensory+abnormalities+in+knee+OA.">Somatosensory abnormalities in knee OA.</a> Rheumatology. 51 (3), 535-543 (2012).</li></ol>

作者没有利益冲突需要披露。

对于获取反映狗的感官阈值的准确数据至关重要，狗在每次测量中都尽可能平静、放松和适当的位置。之前的一项研究指出，测试环境内外因素的约束或分心的躁动会影响狗对 QST 刺激的反应16。如果狗因卧位或约束而变得激动或分心，则应在进行测量之前给狗时间安定下来;没有很快安顿下来的狗应该从测试过程中短暂休息一下。过度焦虑或因测试程序而感到压力的狗可能会表?...

定量感官测试（QST）评估外部应用刺激引起的反应;它用于评估人类和动物的体感系统的功能1。点状压力或深层压力形式的机械刺激作为斜坡刺激应用。感觉阈值被确定为唤起心理物理反应的力量1.热或冷热刺激可用作斜坡刺激或固定强度刺激。感觉阈值确定为对刺激有反应的温度或反应的潜伏期。使用电子冯弗雷麻醉仪或冯弗雷毛丝测量点状压力感觉阈值，使用手持式压力测高仪测量深压力，使用各种接触式热计确定热感觉阈值。
QST提供有关外周和中枢感觉通路功能的信息，可用于评估各种疾病过程中这些感觉通路的改变（可塑性），特别是那些引起慢性疼痛的疾病过程1。迈斯纳微粒检测点状压力，当刺激强度为有害强度时，感觉由非有害水平的 Aβ 传入纤维和 Aδ 传入纤维传递。深压力由帕西尼亚小体检测并由C传入纤维传递，有害热量由鲁菲尼小体检测并由Aδ和C传入纤维传递，有害寒冷由Krause小体检测并由C传入纤维1，2传递。QST可用于检测这些受体和途径的抑制（敏感性降低，感觉减退）和促进（敏感性增加，感觉过敏）。在狗中，QST 已被用于评估继发于急性脊髓损伤3，4，5、小脑扁桃体扁桃体下疝样畸形和脊髓空洞症6、颅交叉韧带断裂5，7 和骨关节炎 （OA） 的感觉阈值改变8，9，10。此外，一些研究使用QST来评估某些镇痛药6，11，12，13和外科手术提供的疼痛缓解14。这些研究为狗的疼痛感机制提供了重要的见解，例如手术后外周和中枢致敏的证据以及导致慢性疼痛状态的疾病，如颅十字韧带断裂和OA。这些信息可以帮助改善狗疼痛的检测和治疗。
狗的机械和热热 QST 验证研究表明，随着时间的推移，正常狗和患有 OA8、9、15、16 慢性疼痛的狗的 QST 结果具有良好的可行性、可重复性和可靠性。然而，一些研究发现冷热和偶尔的冯弗雷QST1，15，17的重复性和可靠性较差。这些研究使用了不同的设备和方法，但提供了证据表明机械和热热QST是测量狗感觉阈值的准确，半定量方法。然而，注意精确的细节，包括测量的设置，对于优化狗的QST至关重要，因此需要标准化的QST协议。Sanchis-Mora等人详细介绍了机械和冷热QST的感觉阈值检查协议（STEP），但狗对冷热QST或研究中使用的最高克力冯弗雷灯丝没有反应遇到了困难17。以下协议提供了狗的机械和热热QST的标准方法;该协议可以评估正常狗或具有影响体感系统的各种疾病过程的狗的感觉阈值。标准化方案的开发可以允许比较研究的结果和数据荟萃分析，以提高QST在兽医学中的效用。

<table><tbody><tr><td>Electronic von Frey anesthesiometer</td><td>IITC Life Science Inc.</td><td>Item # 23931</td><td>Custom made with a 1000g max force load cell</td></tr><tr><td>Medoc Main Station software</td><td>Medoc</td><td>(supplied with TSA-II)</td><td></td></tr><tr><td>SMALGO: SMall Animal ALGOmeter</td><td>Bioseb</td><td>Model VETALGO</td><td></td></tr><tr><td>TSA-II NeuroSensory Analyzer</td><td>Medoc</td><td>DC 00072 TSA-II</td><td>No longer manufactured - new model is TSA-2 with same probes and same function</td></tr></tbody></table>

assessment of sensory thresholds in dogs using mechanical and hot thermal quantitative sensory testing

定量感觉测试 （QST） 用于通过评估对施加的机械和热刺激的反应来评估狗的体感系统功能。QST 用于确定正常狗的感觉阈值，并评估由各种疾病状态引起的外周和中枢感觉通路的改变，包括骨关节炎、脊髓损伤和颅交叉韧带断裂。机械感觉阈值由电子冯弗雷麻醉仪和压力测高仪测量。它们被确定为狗表现出指示有意识刺激感知的反应的力。热热感官阈值是对接触式热化物施加的固定或斜坡温度刺激做出响应的延迟。
遵循一致的方案进行 QST 并注意测试环境、程序和个体研究对象的细节对于获得准确的狗 QST 结果至关重要。尚未详细描述狗QST数据标准化收集的协议。QST 应在安静、无干扰的环境中进行，对狗、QST 操作员和训导员来说都是舒适的。确保狗在每次测量中保持冷静、放松和正确的位置有助于对刺激产生可靠、一致的反应，并使测试过程更易于管理。QST操作员和处理者应该熟悉并熟悉处理狗和解释狗对潜在疼痛刺激的行为反应，以确定测试的终点，减轻压力，并在测试过程中保持安全。

已经进行了机械和热QST以检测研究和客户拥有的狗在各种临床条件下的感觉阈值，包括正常，健康的狗，患有慢性疼痛状况（如OA）的狗，患有急性脊髓损伤的狗，并评估术后疼痛和镇痛药的有效性。尽管在狗身上的QST方面的工作越来越多，但尚未为任何测试方式建立正常的值范围。然而，一些研究已经评估了狗的机械和热QST的可行性和可重复性，显示QST数据是狗感觉阈值的准确测量值<sup class="xr...

Watch this Scientific Journal Video about Assessment of Sensory Thresholds in Dogs Using Mechanical and Hot Thermal Quantitative Sensory Testing at JoVE.com

Assessment of Sensory Thresholds in Dogs Using Mechanical and Hot Thermal Quantitative Sensory Testing

这项工作描述了机械和热定量感官测试的标准协议，以评估狗的体感系统。使用电子冯弗雷麻醉仪、压力测藻计和热接触热敷物测量感觉阈值。

使用机械和热热定量感官测试评估狗的感官阈值

Quantitative sensory testing (QST) is used to evaluate the function of the somatosensory system in dogs by assessing the response to applied mechanical ...

assessment-sensory-thresholds-dogs-using-mechanical-hot-thermal

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

2.1K 次观看.  North Carolina State University. 这项工作描述了机械和热定量感官测试的标准协议，以评估狗的体感系统。使用电子冯弗雷麻醉仪、压力测藻计和热接触热敷物测量感觉阈值。

视频: Assessment of Sensory Thresholds in Dogs Using Mechanical and Hot Thermal Quantitative Sensory Testing

Measurement of Vibration Detection Threshold and Tactile Spatial Acuity in Human Subjects

Tests that allow the precise determination of psychophysical thresholds for vibration and grating orientation provide valuable information about mechanosensory function that are relevant for clinical diagnosis as well as for basic research. Here, we describe two psychophysical tests designed to determine the vibration detection threshold (automated system) and tactile spatial acuity (handheld device). Both procedures implement a two-interval forced-choice and a transformed-rule up and down experimental paradigm. These tests have been used to obtain mechanosensory profiles for individuals from distinct human cohorts such as twins or people with sensorineural deafness.

Here, we present protocols to determine vibration detection thresholds and tactile acuity using psychophysical methods in man.

振动检测阈值和触觉敏锐的空间在人类受试者的测量

Tests that allow the precise determination of psychophysical thresholds for vibration and grating orientation provide valuable information about ...

科研

行为学

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

在大鼠脊髓损伤后的急性和慢性触觉测试

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

医学

A Protocol of Manual Tests to Measure Sensation and Pain in Humans

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study demonstrates a quantitative sensory testing protocol using techniques to measure tactile sensation and pain threshold for pressure and heat using portable and easily accessed equipment. These techniques and equipment are ideal for new laboratories and clinics where cost is a concern or a limiting factor. We demonstrate measurement techniques for the following: cutaneous mechanical sensitivity on the arms and legs (von-Frey filaments), radiant and contact heat sensitivity (with both threshold and qualitative assessments using the Visual Analog Scale (VAS)), and mechanical pressure sensitivity (algometer, with both threshold and the VAS). The techniques and equipment described and demonstrated here can be easily purchased, stored, and transported by most clinics and research laboratories around the world. A limitation of this approach is a lack of automation or computer control. Thus, these processes can be more labor intensive in terms of personnel training and data recording than the more sophisticated equipment. We provide a set of reliability data for the demonstrated techniques. From our description, a new laboratory should be able to set up and run these tests and to develop their own internal reliability data.

The goal of this procedure is to demonstrate a battery of quantitative techniques for sensory and pain measurement in humans. The equipment and techniques described are commonly found in pain clinics or are easy to obtain.

手动测试的协议来衡量感觉和疼痛人类

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study ...

Determining heat and mechanical pain threshold in inflamed skin of human subjects

In a previous article in the Journal of Visualized Experiments we have demonstrated skin microdialysis techniques for the collection of tissue-specific nociceptive and inflammatory biochemicals in humans. In this article we will show pain-testing paradigms that are often used in tandem with microdialysis procedures. Combining pain tests with microdialysis provides the critical link between behavioral and biochemical data that allows identifying key biochemicals responsible for generating and propagating pain.
Two models of evoking pain in inflamed skin of human study participants are shown. The first model evokes pain with aid of heat stimuli. Heat evoked pain as described here is predominantly mediated by small, non-myelinated peripheral nociceptive nerve fibers (C-fibers). The second model evokes pain via punctuated pressure stimuli. Punctuated pressure evoked pain is predominantly mediated by small, myelinated peripheral nociceptive nerve fibers (A-delta fibers). The two models are mechanistically distinct and independently examine nociceptive processing by the two major peripheral nerve fiber populations involved in pain signaling.
Heat pain is evoked with aid of the TSA II, a commercially available thermo-sensory analyzer (Medoc Advanced Medical Systems, Durham, NC). Stimulus configuration and delivery is handled with aid of specific software. Thermodes vary in size and shape but in principle consist of a metal plate that can be heated or cooled at various rates and for different periods of time. Algorithms assessing heat-evoked pain are manifold. In the experiments shown here, study participants are asked to indicate at what point they start experiencing pain while the thermode in contact with skin is heated at a predetermined rate starting at a temperature that does not evoke pain. The thermode temperature at which a subject starts experiencing pain constitutes the heat pain threshold.
Mechanical pain is evoked with punctuated probes. Such probes are commercially available from several manufacturers (von Frey hairs). However, the accuracy of von Frey hairs has been criticized and many investigators use custom made punctuated pressure probes. In the experiments shown here eight custom-made punctuated probes of different weights are applied in consecutive order, a procedure called up-down algorithm, to identify perceptional deflection points, i.e., a change from feeling no pain to feeling pain or vice versa. The average weight causing a perceptional deflection constitutes the mechanical pain threshold.

Algorithms assessing heat and mechanical pain thresholds in experimentally inflamed skin of human study subjects are shown. The two pain testing paradigms independently examine nociceptive processing by the two major peripheral nerve fiber populations transmitting pain, i.e., non-myelinated C fibers and small myelinated A-delta fibers.

确定人类受试者皮肤发炎的热和机械痛阈

In a previous article in the Journal of Visualized Experiments we have demonstrated skin microdialysis techniques for the collection of tissue-specific ...

生物学

Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity to normally innocuous stimuli1. Hyperalgesia and allodynia have been studied in many different rodent models of diabetes mellitus2. However, as stated by B&ouml;lcskei et al, determination of "pain" in animal models is challenging due to its subjective nature3. Moreover, the traditional methods used to determine behavioral responses to noxious thermal stimuli usually lack reproducibility and pharmacological sensitivity3. For instance, by using the hot-plate method of Ankier4, flinch, withdrawal and/or licking of either hind- and/or fore-paws is quantified as reflex latencies at constant high thermal stimuli (52-55 &deg;C). However, animals that are hyperalgesic to thermal stimulus do not reproducibly show differences in reflex latencies using those supra-threshold temperatures3,5. As the recently described method of B&ouml;lcskei et al.6, the procedures described here allows for the rapid, sensitive and reproducible determination of thermal nociceptive thresholds (TNTs) in mice and rats. The method uses slowly increasing thermal stimulus applied mostly to the skin of mouse/rat plantar surface. The method is particularly sensitive to study anti-nociception during hyperalgesic states such as PDN. The procedures described bellow are based on the ones published in detail by Alm&aacute;si et al 5 and B&ouml;lcskei et al 3. The procedures described here have been approved the Laboratory Animal Care and Use Committee (LACUC), Wright State University.

Here, we describe a rapid reliable and simple procedure to determine the lowest temperature at which rats or mice show nocifensive behavior, i.e. the thermal nociceptive threshold (TNT). This method applies a slowly increasing thermal stimulus allowing precise and reproducible estimation of TNTs with minimum, if any, stress to the animals.

糖尿病大鼠的热痛觉阈值的快速测定

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity ...

神经科学

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain perception in rodents. Withdrawal thresholds can be measured first at baseline and then following drug, venom, injury, allergen,&#160;or otherwise evoked inflammation by applying an accurate force on very specific areas of the skin. An electronic von Frey apparatus allows precise assessment of mouse hind paw withdrawal thresholds that are not limited by the available filament sizes in contrast to classical von Frey measurements. The ease and rapidity of measurements allow for incorporation of assessment of tactile sensitivity outcomes in diverse models of rapid-onset inflammatory and neuropathic pain as multiple measurements can be taken within a short time period. Experimental measurements for individual rodent subjects can be internally controlled against individual baseline responses and exclusion criteria easily established to standardize baseline responses within and across experimental groups. Thus, measurements using an electronic von Frey apparatus represent a useful modification of the well-established classical von Frey filament-based assays for rodent mechanical allodynia that may also be applied to other nonhuman mammalian models.

Electronic von Frey measurements are an effective and improved alternative to classical von Frey filaments, as they allow rapid and precise measurement of changes in rodent mechanical withdrawal thresholds to continuously applied pressure stimuli before and after an inflammatory event.

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain ...

Back Mechanical Sensitivity Assessment in the Rat for Mechanistic Investigation of Chronic Back Pain

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, animal models are needed to examine the mechanisms and effectiveness of novel therapies from a translational perspective. Several rodent models of back pain are used in current investigations. Surprisingly, however, no standardized behavioral test was validated to assess mechanical sensitivity in back pain models. This is critical to confirm that animals with presumed back pain present local hypersensitivity to nociceptive stimuli, and to monitor sensitivity during interventions designed to relieve back pain. The objective of this study is to lay down a simple and accessible test to assess mechanical sensitivity in the back of rats. A test cage was fabricated specifically for this method; length x width x height: 50 x 20 x 7 cm, having a stainless-steel mesh on the top. This test cage allows the application of mechanical stimuli to the back. To perform the test, the back of the animal is shaved in the region of interest, and the test area is marked to repeat the test on different days, as needed. The mechanical threshold is determined with Von Frey filaments applied to the paraspinal muscles, utilizing the up-down method described previously. The positive responses include (1) muscle twitching, (2) arching (back extension), (3) rotation of the neck (4) scratching or licking the back, and (5) escaping. This behavioral test (Back Mechanical Sensitivity (BMS) test) is useful for mechanistic research with rodent models of back pain for the development of therapeutic interventions for the prevention and management of back pain.

To develop novel therapeutic interventions for the prevention and management of back pain, animal models are required to examine the mechanisms and effectiveness of these therapies from a translational perspective. The present protocol describes the BMS test, a standardized method to assess back mechanical sensitivity in the rat.

大鼠背部机械敏感性评估用于慢性背痛的机制研究

Low back pain is the leading cause of disability worldwide, with dramatic personal, economic, and social consequences. To develop novel therapeutics, ...

Assessment of Nerve Injury-Induced Mechanical Hypersensitivity in Rats Using an Orofacial Operant Pain Assay

Pain has sensory and affective components. Unlike traditional, reflex-based pain assays, operant pain assays can produce more clinically relevant results by addressing the cognitive and motivational aspects of pain in rodents. This paper presents a protocol for assessing mechanical hypersensitivity following chronic constriction injury of the infraorbital nerves (CCI-ION) in rats using an orofacial operant pain system. Before CCI-ION surgery, rats were trained in an orofacial pain assessment device (OPAD) to drink sweetened condensed milk while making facial contact with the metal spiked bars and lick-tube.
In this assay, rats can choose between receiving milk as a positive reinforcer or escaping an aversive mechanical stimulus that is produced by a vertical row of small pyramid-shaped spikes on each side of the reward access hole. Following 2 weeks of training in the OPAD and before the CCI-ION surgery, baseline mechanical sensitivity data were recorded for 5 days for each rat during a 10 min testing session. During a session, the operant system automatically records the number of reward bottle activations (licks) and facial contacts, contact duration, and latency to the first lick, among other measures.
Following baseline measurements, rats underwent either CCI-ION or sham surgery. In this protocol, mechanical hypersensitivity was quantified by measuring the number of licks, latency to the first lick, the number of contacts, and the ratio of licks to facial contacts (L/F). The data showed that CCI-ION resulted in a significant decrease in the number of licks and the L/F ratio and an increase in the latency to the first lick, indicating mechanical hypersensitivity. These data support the use of operant-based pain assays to assess mechanical pain sensitivity in preclinical pain research.

This protocol describes the assessment of mechanical hypersensitivity in a rat model of neuropathic orofacial pain using an operant-based orofacial pain assessment device.

使用口面部操作性疼痛测定法评估大鼠神经损伤诱导的机械超敏反应

Pain has sensory and affective components. Unlike traditional, reflex-based pain assays, operant pain assays can produce more clinically relevant results ...

Dynamic Quantitative Sensory Testing to Characterize Central Pain Processing

Central facilitation and modulation of incoming nociceptive signals play an important role in the perception of pain. Disruption in central pain processing is present in many chronic pain conditions and can influence responses to specific therapies. Thus, the ability to precisely describe the state of central pain processing has profound clinical significance in both prognosis and prediction. Because it is not practical to record neuronal firings directly in the human spinal cord, surrogate behavior tests become an important tool to assess the state of central pain processing. Dynamic QST is one such test, and can probe both the ascending facilitation and descending modulation of incoming nociceptive signals via TS and CPM, respectively. Due to the large between-individual variability in the sensitivity to noxious signals, standardized TS and CPM tests may not yield any meaningful data in up to 50% of the population due to floor or ceiling effects. We present methodologies to individualize TS and CPM so we can capture these measures in a broader range of individuals than previously possible. We have used these methods successfully in several studies at the lab, and data from one ongoing study will be presented to demonstrate feasibility and potential applications of the methods.

By assessing pain in response to repetitive or different types of standardized stimuli, dynamic quantitative sensory testing (QST) can reveal changes in the central processing of pain. We present methods to optimize and individualize two dynamic QST measures: temporal summation (TS) and conditioned pain modulation (CPM).

动态定量感觉测试表征中央疼痛处理

Central facilitation and modulation of incoming nociceptive signals play an important role in the perception of pain. Disruption in central pain ...

Establishing a Mouse Model of a Pure Small Fiber Neuropathy with the Ultrapotent Agonist of Transient Receptor Potential Vanilloid Type 1

Patients with diabetes mellitus (DM) or those experiencing the neurotoxic effects of chemotherapeutic agents may develop sensation disorders due to degeneration and injury of small-diameter sensory neurons, referred to as small fiber neuropathy. Present animal models of small fiber neuropathy affect both large- and small-diameter sensory fibers and thus create a neuropathology too complex to properly assess the effects of injured small-diameter sensory fibers. Therefore, it is necessary to develop an experimental model of pure small fiber neuropathy to adequately examine these issues. This protocol describes an experimental model of small fiber neuropathy specifically affecting small-diameter sensory nerves with resiniferatoxin (RTX), an ultrapotent agonist of transient receptor potential vanilloid type 1 (TRPV1), through a single dose of intraperitoneal injection, referred to as RTX neuropathy. This RTX neuropathy showed pathological manifestations and behavioral abnormalities that mimic the clinical characteristics of patients with small fiber neuropathy, including intraepidermal nerve fiber (IENF) degeneration, specifically injury in small-diameter neurons, and induction of thermal hypoalgesia and mechanical allodynia. This protocol tested three doses of RTX (200, 50, and 10 &#181;g/kg, respectively) and concluded that a critical dose of RTX (50 &#181;g/kg) is required for the development of typical small fiber neuropathy manifestations, and prepared a modified immunostaining procedure to investigate IENF degeneration and neuronal soma injury. The modified procedure is fast, systematic, and economic. Behavioral evaluation of neuropathic pain is critical to reveal the function of small-diameter sensory nerves. The evaluation of mechanical thresholds in experimental rodents is particularly challenging and this protocol describes a customized metal mesh that is suitable for this type of assessment in rodents. In summary, RTX neuropathy is a new and easily established experimental model to evaluate the molecular significance and intervention underlying neuropathic pain for the development of therapeutic agents.

This study establishes an experimental model of pure small fiber neuropathy with resiniferatoxin (RTX). A unique dose of RTX (50 &#181;g/kg) is optimal for developing a small fiber neuropathy model that mimics patient characteristics and could help investigate the nociceptive molecular significance underlying neuropathic pain.

利用瞬态受体电位辣椒1型 Ultrapotent 激动剂建立单纯小纤维神经病变的小鼠模型

Patients with diabetes mellitus (DM) or those experiencing the neurotoxic effects of chemotherapeutic agents may develop sensation disorders due to ...