该项目由工程和物理科学研究理事会 EPSRC (EP/K503745/1)、国家卫生研究所 (NIHR) 生物医学研究中心 (BRC272/HI/JG/101440) 和伦敦大学学院的变革制造商提供资金。

该议定书遵循伦敦大学学院人类研究伦理委员会的指导方针。
警告: 本协议包含电气危害和烧伤危险 (烙铁);请在尝试此协议之前读取两个内容。此协议包括将设备连接到皮肤。确保在任何时间, 在皮肤和电源之间有一个路径。不要接触烙铁的元素。用镊子或夹子将电线加热。使用时保持清洁海绵湿润。在不使用时, 始终将烙铁退回其支架。不要把它放在工作台上。关闭设备并在不使用时拔下插头。
1. 组装基本组件
注意:图 3提供了协议步骤的高级概述。
<ol><li>要建立仿生的响片 MK I, 将生物信号传感器屏蔽插入到微控制器中, 并将肌电信号的缆线螺钉插入屏蔽的 E、M 和接地螺钉端子 (请参见图 4)。继续执行步骤1.6。</li>
	<li>
要构建 MK II, 请在 + 的肌肉传感器中放置一行三头针,-和 SIG 孔 (请参阅上面的图 5) 和下面的焊料。
	<ol>
<li>弯曲的头针90°与一对 plyers 一半的针脚。这安置别针在正确位置为案件。</li>
		<li>如果使用诱拐食指肌肉作为输入, 继续步骤 1.3, 如果不移动到步骤1.4。</li>
	</ol>
</li>
	<li>
从肌肉传感器中取出短的黑色参考线。
	<ol>
<li>用线切割器切断三肌电图电缆, 从手腕到手的后部。用铁丝剥线剥去三肌电信号的末端。</li>
		<li>将黑线的剥离端放入 R 孔, 将蓝线插入 E 孔, 并将红色导线插入肌肉传感器的 M 孔 (图 5)。在肌肉传感器的底部焊接电线。移到步骤1.5。</li>
	</ol>
</li>
	<li>
将两个电极垫夹入肌肉传感器的下侧, 并将一个电极插入黑色参考线的连接器中。
	<ol>
<li>用电极垫将肌肉传感器粘在选定的肌肉上, 并将黑色参考电极放置在适当的位置。</li>
	</ol>
</li>
	<li>
将8单核多线程线切割成长度和带末端: 5 短 (7 厘米) 电线运行从单片机到控制板 (红色, 黑色, 绿色, 白色和蓝色) 和3更长 (约12厘米, 但依赖于手腕尺寸) 电线 (红色, 黑色和绿色)从手腕的一端跑到另一侧。 注: 如果将肌肉传感器放置在不同的肌肉上, 请确保较长的电线将从肌肉传感器站点运行到无线微控制器站点。<ol>
<li>将导线放入微控制器中准备焊接: 2 条红线进入3V 孔, 2 条黑线进入接地孔, 长青线进入 A0 孔, 短蓝线成2孔, 长白线进入3孔, 并将短的绿线插入到5孔中。在微控制器的底部焊接电线。</li>
		<li>将3条长导线的另一端焊至3头插座, 顺序为: 红色、黑色、绿色。请参见图 5。如果不使用诱拐食指肌肉继续到步骤2。</li>
	</ol>
</li>
	<li>
将肌电信号传感器垫放在手上, 如图 6所示, 其中两个电极位于食指肌的一端, 一个肌电传感器垫位于手的背部中部。
	<ol>
<li>将电极垫夹入肌肉传感器电缆的接头端 (推合)。蓝色和红色电极夹在肌肉上方, 黑色电极夹在手的背面。</li>
	</ol>
</li>
</ol>2. 测试肌电信号输出
<ol><li>从参考部分的链接14之后, 下载生物信号屏蔽库。解压它并将其放在集成开发 (IDE) 库文件夹中 (通常在文档/Arduino/库中找到)。继续执行步骤2.3。如果生成 MK II, 请继续执行步骤2.2。</li>
	<li>按照15的说明, 将微控制器主板添加到 IDE 中。</li>
	<li>下载 "ThresholdTest" 的 mk i 或 "BLEThresholdTest" 和 "BluefruitConfig", 用于 mk II 并在 IDE 软件 (补充文件) 中打开。</li>
	<li>从电源上拔下笔记本电脑, 然后, 然后, 通过通用串行总线 (USB) 电缆将微控制器插入笔记本电脑。</li>
	<li>将相关的阈值测试版本上传到微控制器上, 然后打开串行监视器 (工具 > 串行监视器)。现在将显示肌电信号的输出。</li>
	<li>移动食指从一边到另一边并且移动手在附近, 不用移动食指。记下每个案例中显示的值。 注意: 当使用 MK II 确保电缆不移动, 因为它是非常敏感的噪音产生这种方式。</li>
	<li>选择一个值高于手移动时所看到的数值, 但在手指从一侧移到另一侧时所看到的下方。记下这个值。 注意: 该值是选定的, 以便该设备只会被激活的手指有目的的运动。这是阈值触发器数值, 将激活该设备的值。肌肉传感器有一个增益设置, 可以手动改变, 如果阈值是很难找到。电极可能需要更换。如果使用食指肌, 将增益设定为最小值作为起始点。增益设置是改变由电位器上的肌肉传感器标记的增益, 这可以改变一个小平头螺丝刀。</li>
</ol>3. 测试阈值
<ol><li>下载 "BoomTest" 的 mk i 或 "BLEBoomTest" 和 BluefruitConfig h 为 mk II, 并在 IDE 软件中打开它。</li>
	<li>通过将 "PLACE_YOUR_THRESHOLD_TRIGGER_VALUE_HERE" 替换为前面在步骤2.8 中确定的阈值触发器, 来编辑所提供的代码。这是 mk I 和 II. 代码47行代码的37行</li>
	<li>将 BoomTest 的正确版本上传到微控制器上, 然后打开串行监视器 (工具 > 串行监视器)。</li>
	<li>移动手 (不移动食指从一边到另一边);在串行输出上没有看到任何东西。</li>
	<li>将食指从一边移动到另一侧;"繁荣" 这个词出现了。 注意: 如果 "BOOM" 出现在错误的时间或根本不显示, 请检查连接并移回步骤2.7。</li>
</ol>4. 3D 打印 MK II 例
<ol><li>如果生成 MK II, 请为案例的所有5组件下载 stl 文件 (请参见图 7中的所有5部分)。按任何首选方法打印案例的各个部分。继续执行步骤5.2。如果建立 MK I, 继续5节。 注: 该案例已成功印出熔融沉积模拟16 (差分) 和光刻机打印机17。</li>
</ol>5. 焊接控制板
注意: 如果生成 MK II, 请继续执行步骤5.2。
<ol><li>
放置一排两个头引脚, 五10ω电阻器, 滑动开关和两个按钮开关的组成部分, 如图 8a;然后把它们焊在板子的底部。
	<ol>
<li>按照图 8a上的灰色线, 用工艺刀进行切片, 以打破带状板上的铜轨。这使得各个轨道在整个电路板上具有多个功能。</li>
		<li>切割7线 (黑色, 红色, 蓝色, 橙色, 白色, 褐色和黄色) 的正确长度与一个线切割机, 使他们将运行从前臂到上臂 (约30厘米)。切割7厘米的红线, 黑色线3厘米和橙色, 蓝色线的4厘米。</li>
		<li>用铁丝剥线剥去电线的两端。</li>
		<li>将导线放在控制板中, 按照图 9中所示的电路图进行布线;将电线焊在下面的地方。</li>
		<li>将长的红色和黑色的导线焊接到一对头针上, 然后再将其他长的导线焊接到一条标头针脚中, 顺序为: 蓝色、橙色、白色、褐色、黄色。</li>
		<li>将无线模块的5V 和地线针脚焊接到控制板上的接头针脚上。</li>
		<li>将短的橙色线焊接到无线通信模块的2针和短的蓝线以引脚3。</li>
	</ol>
</li>
	<li>
放置三10ω电阻器、滑动开关和两个按钮开关, 如图 10a 所示, 并将其焊接到主板底部。
	<ol>
<li>按照图 10A上的灰色线, 通过用工艺刀进行切片, 打破条板上的铜轨。这使得轨道在电路板上有多个功能。</li>
		<li>切断以前焊接到微控制器上的导线与一个线切割机, 使他们可以运行通过的微控制器外壳的中间层到控制板, 而不停止的情况下关闭 (图 10B)。</li>
		<li>按照电路图 (图 11) 将导线放在控制板上。把电线焊到位。继续执行步骤6.2。</li>
	</ol>
</li>
</ol>6. 组装和更新单片机
<ol><li>重新组装的仿生的响片, 连接头连接器从控制板电线到微控制器和生物信号盾 (5V 和地线上 MK I, 引脚22-30 在 MKII)。将电池连接到微控制器。请参见图 12。转到步骤6.3。</li>
	<li>重新组装仿生遥控器, 将接头从单片机连接到肌肉传感器 (绿色导线到 SIG)。请参见图 13。</li>
	<li>通过 USB 电缆将微控制器连接到笔记本电脑。</li>
	<li>下载 "BionicClicker" 或 "BLEBionicClicker 和 BluefruitConfig", 然后在 IDE 软件中打开它。</li>
	<li>编辑代码并将 "PLACE_YOUR_THRESHOLD_TRIGGER_VALUE_HERE" 替换为2.7 步中确定的阈值触发器 (mk II 的代码的59行中的 "第一行", 该代码中的83线, 用于第二部分)。 注意: 通过编辑代码的47 行, 可以更改 MK II 设备在通过无线连接时出现的名称。使用替代标题替换 "仿生遥控器 MK II"。</li>
	<li>卸下 USB 电缆以断开微控制器与便携式计算机的连接。</li>
</ol>7. 将设备连接到计算机
<ol><li>如果使用 MK I, 请按照制造商指南18的说明将无线模块与设备配对。如果使用 MK II, 则通过无线键盘与正在使用的计算机连接的过程无线连接到设备。</li>
</ol>8. 测试遥控器
<ol><li>
打开一些打字软件, 并输入一些文字, 如 ' Lorem ipsum 忧伤坐 amet '。这使得印刷机被视为测试是否发送和接收这些命令。 注意: 如果电池电量不足, 设备可能会发出不稳定的行为;总是使用新电池。<ol>
<li>按 "手动前进" 按钮可看到光标向前移动, 手动后退按钮可看到光标向后移动。抬起食指也向前移动。</li>
	</ol>
</li>
	<li>要用演示软件测试遥控器, 请抬起食指以进行幻灯片的进度。 注意: "覆盖开关" 打开和关闭 "肌电信号" 功能, "手动前进" 和 "后退" 按钮在两种情况下都会前进并后退幻灯片。</li>
</ol>9. 安装遥控器
注意: 如果生成 MK II 移动到步骤9.2。
<ol><li>
如果建立 MK I, 削减双面钩和线圈的材料与剪刀, 使其适合舒适的手腕周围。确保环面朝内, 不划伤手腕。
	<ol>
<li>切割双面钩和线圈的材料, 使其适合舒适地在上臂, 再次确保环面朝内。</li>
		<li>将双面钩和环条切至微控制器 (10 cm x 5 cm) 和控制板 (2.5 cm x 6.4 厘米) 的大小。切一条将紧紧围绕电池 (4 厘米 x 12 厘米)。</li>
		<li>使用胶枪, 胶水的线圈一侧的小条的底部的微控制器和底部的控制板。</li>
		<li>将控制板连接到腕带上。将微控制器和电池连接到上臂皮带上。</li>
		<li>插头的所有内容: 9 伏电池插头进入微控制器与 PP3 连接器。微控制器和电子卫生盾通过焊接的导线连接到控制板。 注: MK I 现已完成。</li>
	</ol>
</li>
	<li>
如果建立 MK II, 削减双面钩和循环材料35毫米宽, 足够长的时间包在手腕 (约22厘米较小的手腕)。
	<ol>
<li>滑动钩和循环材料通过剪辑在底部的情况下。确保环面朝内, 不划伤手腕。</li>
		<li>将焊接到微控制器上的导线插入到肌肉传感器上的公头针脚上, 并将电极插入肌电信号中, 并将其推入。 注: MK II 现已完成。请参见图 14。</li>
	</ol>
</li>
</ol>

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Arduino IDE Setup Available from: <a target="_blank" href="https://learn.adafruit.com/add-boards-arduino-v164/setup">https://learn.adafruit.com/add-boards-arduino-v164/setup</a> (2017)</li><li>. Ultimaker 2+ Available from: <a target="_blank" href="https://ultimaker.com/en/products/ultimaker-2-plus">https://ultimaker.com/en/products/ultimaker-2-plus</a> (2017)</li><li>. Form 2 Available from: <a target="_blank" href="https://formlabs.com/3d-printers/form-2/">https://formlabs.com/3d-printers/form-2/</a> (2017)</li><li>. Pairing to Bluefruit Available from: <a target="_blank" href="https://learn.adafruit.com/introducing-bluefruit-ez-key-diy-bluetooth-hid-keyboard/pairing-to-bluefruit">https://learn.adafruit.com/introducing-bluefruit-ez-key-diy-bluetooth-hid-keyboard/pairing-to-bluefruit</a> (2017)</li></ol>

作者没有什么可透露的。

MK II 的饱和度, 当用于诱拐食指是较少的问题比它5月1日出现。小心放置的电极和正确的增益设置停止这是一个问题时, 设备是作为一个响片。除非有兴趣准确记录诱拐者指数的活动, 否则这不可能是一个问题。在其他肌肉被确定后, 没有任何饱和被发现。假阴性与 MK II 是由于困难的选择适当的阈值时, 使用诱拐食指。与更大的肌肉区别在大小之间肌肉的 non-purposeful 活化作用和有目的紧张肌肉是更大的, 允许选择一个门限点进一步从假正面和假阴性点.在特别小的手食指肌可能是太小的电极被正确地安置 (虽然与更小的电极垫这可能可能被解决)。
由于多种原因, MK II 的电池寿命较长很有用。首先, MK I 装置在使用45分钟后开始发生不稳定的行为, 因此不能用于更长的演示。其次, multi-hour 电池寿命, MK II 可以被认为是一个有用的设备的控制输入, 并只增加了一个小的物理电池的大小, 它可以作为全天候监测设备。无线微控制器有6模拟输入和13数字输入;这意味着该设备可以接受来自多个肌肉传感器的信号, 在控制输入中创建具有更多自由度的设备。还应该指出, 肌肉传感器可以取代任何生物传感器与模拟输出创建一个设备, 使用其他生物信号作为输入。该设备的代码也可以很容易地修改, 以改变其功能。对设备的软件和硬件的更改允许对设备进行简单而多样的修改。
该设备目前的一个限制是, 不能以高数据速率以无线方式发送肌电信号输出, 因为这会使无线微控制器缓冲区过载。另一个限制是, 该技术使用诱拐指数作为输入, 并由于肌肉非常小, 在手的电极间距几乎重叠;如果某人有特别小的手, 它可能不可能正确地安置电极在这个肌肉。
当涉及到潜在研究项目的灵活性时, 该设备比较昂贵的设备有几个优点。这是低成本: 设备成本80美元和额外的肌电信号通道只花费35美元, 使其适合较小或学生项目。它易于自定义, 软件可以很容易地编辑, 并为其他硬件的输入改变。它有一个小尺寸, 所以一个人穿它不需要携带笨重或笨重的设备。它还会显示为其他设备的无线键盘, 因此可以方便地与任何兼容的无线设备集成。该设备已被纳入一个辅助设备, 将在不久的将来出版。
由于 MK II 的定制的规模和易用性, 它已经被考虑作为一个无线肌电信号模块和无线触发机制纳入几个研究项目。它也被用来作为一个实验课的基础上, 一个硕士学生的课程。我们想对该设备进行的主要改进是提高无线传输速率;目标是实现10赫兹, 这是否将通过硬件或软件来完成, 还有待确定。
协议中最关键的步骤是步骤2.6 和 2.7: 阈值触发器的选择。在步骤2.6 中, 需要特别注意肌电信号的移动, 因为它们可以充当天线和产生运动伪影;但是, 如果这些保持静止, 这不是一个问题。在步骤2.7 中, 如果所选值太高, 则会导致错误的负数。如果此值过低, 则会导致误报。在诱拐食指的情况下, 很难找到一个不会导致偶然的假阴性的值, 虽然与较大的肌肉, 这似乎不是一个问题。如果找到正确的值仍然是一个问题, 增益可以纠正通过设置它的最小值, 并增加它, 直到非和激活之间的一个很大的差异, 通过串行读出, 值保持低于点饱和.
总体上 mk II 是一个相当大的改进, 在 mk i 作为一个潜在的研究设备, 虽然因为 mk 我有一个更强的视觉冲击力, 它很可能仍然被用于未来的公众参与事件。

渴望康复工程和辅助技术中心调查在相关领域的不同领域适用和可转让的技术, 包括但不限于中风、肌肉萎缩症、截肢, 人口老龄化和专业技能培训。中心所涉及的研究领域之一是 neuroprosthesis。在控制 neuroprosthetic 武器的许多技术中, 肌电信号是控制系统的最常见的输入1,2,3,45。与其他控制系统7相比, 这在很大程度上是因为它的易用性和可承受能力。最近开发的 3 d 印刷假体, 如 Ada 手, 在使用这种类型的控制8,9,10时只能花费1000美元。然而, 在试图向公众展示这种系统时, 如果没有截肢的人的帮助, 就没有简单的办法。
为了提高这一领域的研究活动对公众的认识, 研制了一种仿生的响片演示装置。使用基于对象的演示是非常重要的, 因为它吸引了注意并加速了11所教授的主题的学习和理解。我们的设备不仅有助于教授肌电图的概念, 还能提高对现代技术发展的认识。此外, 它还鼓励年轻一代在科学、技术、工程和数学 (词干) 领域内选择研究。
仿生的选择 MK I 是使用现成的零件, 已经在实验室内使用。它包括一个微控制器, 一个生物信号盾12, 电极, 一个控制板, 一个无线通信板和一个 9 V 电池。该装置的功能是拾取的活动食指肌肉位于食指和拇指之间。它通过模仿键盘并在达到预选阈值时发送 "右按键" 来触发幻灯片更改。控制板允许手动发送 "右" 和 "左" 按键 (进度幻灯片和后退幻灯片), 如果在现场演示中出现问题, 也可以使用它来覆盖肌电信号输入。
作为医学物理和生物医学工程系公共参与活动的一部分, 我们向公众展示了仿生的遥控器。它得到了观众的热烈响应, 并产生了开始几个合作的兴趣。在最初的设备的成功之后设计了第二个版本的设备。
设计第二个设备的目的是制造比第一个设备更便宜、更不繁琐和更可定制的设备。该设备的目的是设计一些很容易被修改的学生项目, 并将其便宜地并入现有项目中。此设备的主要优势在于其他可用的1、2、3、4、5、6是其易用性、小尺寸和低成本。虽然仿生的响片装置可能没有其他研究装置的分辨率, 比如触发装置, 但它们已经足够好了。MK II 将是任何使用肌电信号阈值触发设备 (如假肢控制器或辅助设备) 的系统的理想基础。
该设计是基于一个无线功能的微控制器和肌肉传感器。它还包括一个3.7 伏150毫安的锂聚合物电池, 一个手动控制板和一个 3 d 打印的情况下。图 3显示了设计之间的差异的概览。MK II 设计具有与原始设备相同的基本功能, 但对于新的应用 (如无线肌电信号监控) 具有显著的潜在功能。

<table border="0" cellpadding="0" cellspacing="0" width="1034">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">For the Mark I</td>
			<td style="width:77px;"></td>
			<td style="width:308px;"></td>
			<td style="width:407px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Laptop</td>
			<td></td>
			<td></td>
			<td>Any laptop with USB</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">USB B cable</td>
			<td></td>
			<td></td>
			<td>From laptop to USB-B connection on Arduino</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Soldering Station</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Solder</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Hot glue gun</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Hot glue gun glue</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Items</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Small Single-Core Multi-thread Wires</td>
			<td></td>
			<td></td>
			<td>Black, Red, Yellow, Brown, Orange White, Blue,</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Arduino MEGA 2560</td>
			<td>Arduino</td>
			<td>Arduino MEGA 2560</td>
			<td>(Geniuno MEGA 2560 outside US)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">E-Health Shield v2.0</td>
			<td>Cooking Hacks</td>
			<td>e-Health Sensor Shield V2.0 for Arduino, Raspberry Pi and Intel Galileo [Biometric / Medical Applications]</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">EMG cables</td>
			<td>Cooking Hacks</td>
			<td>Electromyography Sensor (EMG) for e-Health Platform [Biometric / Medical Applications]</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">EMG Electrodes</td>
			<td>Sparkfun</td>
			<td>SEN-12969</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">9V battery</td>
			<td></td>
			<td></td>
			<td>Any</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Power cable</td>
			<td></td>
			<td></td>
			<td>PP3 9v connector with jack</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bluefruit EZ-KEY HID</td>
			<td>Adafruit</td>
			<td align="right">1535</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">strip board</td>
			<td>Amazon.co.uk</td>
			<td>Small Stripboard 25 X 64mm Pack of 3</td>
			<td>any similiar stripboard 2.54mm pitch 7x25</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">push button switch</td>
			<td></td>
			<td>COM-00097</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">slide switch</td>
			<td>amazon.com</td>
			<td>20 Pcs On/Off/On DPDT 2P2T 6 Pin Vertical DIP Slide Switch 9x4x3.5mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">resistors</td>
			<td></td>
			<td>COM-11508</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Double sided Velcro</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Break Away Headers - Straight</td>
			<td>Sparkfun</td>
			<td>PRT-00116</td>
			<td>2, 2 and 5 needed</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">For the Mark II</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Laptop</td>
			<td></td>
			<td></td>
			<td>Any laptop with USB connection</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">USB micro cable</td>
			<td></td>
			<td></td>
			<td>From laptop to USB micro (standard phone connector style)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Soldering Station</td>
			<td></td>
			<td></td>
			<td>Any</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Solder</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Items</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Small Single-Core Multi-thread Wires</td>
			<td></td>
			<td></td>
			<td>Black, Red, Green, White, Blue,</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Feather BLE 32U4</td>
			<td>Adafruit</td>
			<td align="right">2829</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">MyoWare</td>
			<td>Sparkfun</td>
			<td>SEN-13723</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">EMG cables</td>
			<td>Sparkfun</td>
			<td>CAB-12970</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">EMG electrodes</td>
			<td>Sparkfun</td>
			<td>SEN-12969</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">3.7 V LiPo</td>
			<td>Adafruit</td>
			<td align="right">1317</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Strip Board</td>
			<td>amazon.co.uk</td>
			<td>Small Stripboard 25 X 64mm Pack of 3</td>
			<td>2.54 pitch 7x9 rows</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Push Button switch</td>
			<td></td>
			<td>COM-00097</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">slide switch</td>
			<td>amazon.com</td>
			<td>20 Pcs On/Off/On DPDT 2P2T 6 Pin Vertical DIP Slide Switch 9x4x3.5mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">resistors</td>
			<td></td>
			<td>COM-11508</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">3D printed parts</td>
			<td></td>
			<td></td>
			<td>Can be 3D printed yourself or printed from a website</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Double sided Velcro</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Break Away Headers - Straight</td>
			<td>Sparkfun</td>
			<td>PRT-00116</td>
			<td>3 pins needed</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Female Headers</td>
			<td>sparkfun</td>
			<td>PRT-00115</td>
			<td>3 pins needed</td>
		</tr>
	</tbody>
</table>

the bionic clicker mark i &amp; ii

在这篇手稿中, 我们提出了两个 ' 仿生的 ' 系统, 第一个设计, 以演示肌电图 (肌电信号) 为基础的控制系统的教育目的和第二个研究目的。基于肌电信号的控制系统拾取肌肉激活产生的电信号, 并将这些信号作为控制器的输入。肌电控制器广泛应用于假肢控制四肢。
"标记 i" (MK i) "遥控器" 允许佩戴者通过提高其食指来改变演示文稿的幻灯片。它是围绕一个单片机和生物信号屏蔽建立的。它引起了公众和研究界的极大兴趣。
此处介绍的标记 ii (MK ii) 设备设计为更便宜、更时尚、更可定制的系统, 可方便地修改和直接传输肌电信号数据。它是使用无线功能的单片机和肌肉传感器构建的。

mk II 比 mk I 设备更实惠、可定制且不那么繁琐。整个 MK II 成本仅略高于生物信号盾 (75 美元)。该设备是显着较小的坐在手腕上, 而不是手臂和无线微控制器可能同时支持从6肌肉传感器的输入。mk I 设备的功能电池寿命仅在1小时以下, 使用9伏550毫安电池, 而 mk II 装置的功能电池寿命 (当用作遥控器时) 大约8小时使用3.7 伏150毫安电池;有关设备的比较, 请参见表 1 。
仿生的响片 MK II 可以有一个问题时, 使用的诱拐食指: 放大器可以饱和, 并采取超过一秒钟的放电 (参见图 15)。仔细放置电极并正确设置增益可以克服这个问题。这不发生与仿生的响片 MK I 或任何其他常用肌肉的肌电信号。
在校准设备以找到阈值触发价值的同时, 可以观察到许多不同的值。它们分成三范围: 当手静止时的值, 手移动时的值, 手指移动时的值。表 2显示每个范围中的记录值;为固定和手移动的范围, 最大值显示和为手指紧张的范围最小值显示。阈值被选择在手移动值的上方和手指的紧度值之下。接近手移动范围的值增加了假阳性的几率, 减少了假阴性的几率, 而接近手指拉紧范围的值则有相反的效果。
这两种装置都在测试假阴性和假阳性时, 紧张的诱拐指数肌肉。当设备未在肌肉拉紧时触发滑动变化时, 记录了假阴性, 如果在没有拉紧发生的情况下滑动发生变化, 则记录假阳性。这两种设备都没有出现假阳性的问题, 虽然 MK II 设备经历了偶尔的假阴性 (少于5% 的时间)。在操作的前45分钟内, MK I 设备不会出现误报或负片, 但假底片的数量会迅速增加, 直到50分钟到1小时 (参见表 3)。
结果表明, 该装置在其所述目标上取得了成功。表 1显示 mk II 更便宜, 而且比 mk i.表 2和表 3的灵活性更高, 显示该设备按预期的功能运行, 并且可以用作 EMG-based 触发器设备。图 15显示了在使用诱拐指数肌肉时可能出现的问题: 这不是大多数肌肉出现的问题, 可以通过改变增益来修正。虽然这些设备有一些问题, 但它们足以用于预期的用途。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55705/55705fig1.jpg"> 图 1:仿生遥控器 MK i 。这显示了仿生的响片 MK I 及其所有组件安装在左臂上。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55705/55705fig2.jpg"> 图 2:阻止设备图.每个框代表设备的一个单独的部分;每个框中的功能是该部分作为设备的一部分。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/55705/55705fig3.jpg"> 图 3: 生成设备的步骤.包含协议每一步骤的高级概述的流程图。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/55705/55705fig4.jpg"> 图 4: 初始 MK I 组件.单片机与生物信号屏蔽和电极电缆。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/55705/55705fig5.jpg"> 图 5:初始 MK II 程序集.单片机与肌肉传感器和焊接连接。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/55705/55705fig6.jpg"> 图 6: 电极放置.此图显示了使用食指时电极在手上的正确位置。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/55705/55705fig7.jpg"> 图 7: MK II 机箱部件.MK II 案件的部分准备在影印打印机打印。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/55705/55705fig8.jpg"> 图 8: MK I 控制电路.(a) 电路板从上面 (灰色标记的地方板有接触打破了在底部)。(b) 已完成电路板。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig8large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 9" class="xfigimg" src="/files/ftp_upload/55705/55705fig9.jpg"> 图 9: MK I 控制电路板电路图.用于 MK I 控制板的电路图, 显示电阻、开关和导线之间的连接。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig9large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 10" class="xfigimg" src="/files/ftp_upload/55705/55705fig10v2.jpg"> 图 10: MK I 控制电路.(a) 控制板从上面 (灰色标记的地方板有接触打破了在底部)。(b) 已完成电路板<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig10v2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 11" class="xfigimg" src="/files/ftp_upload/55705/55705fig11.jpg"> 图 11:MK II 控制板电路图.用于 MK I 控制板的电路图, 显示电阻、开关和导线之间的连接。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig11large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 12" class="xfigimg" src="/files/ftp_upload/55705/55705fig12.jpg"> 图 12: 组装的 MK i 。这显示了 MK I 装置的所有组件, 然后安装在手臂上。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig12large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 13" class="xfigimg" src="/files/ftp_upload/55705/55705fig13.jpg"> 图 13: 组装遥控器 MK II.(a) 将微控制器放在机箱的底部。(b) 将电池放在中间上并盖上盖子。(c) 将肌肉传感器置于其机箱中, 并将其置于盖子上。(d) 将微控制器连接到肌肉传感器, 并将电池连接到微控制器。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig13large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 14" class="xfigimg" src="/files/ftp_upload/55705/55705fig14.jpg"> 图 14: 完成的仿生遥控器 MK II.(a) 在钩子和环带上。(b) 在手腕上。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig14large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 15" class="xfigimg" src="/files/ftp_upload/55705/55705fig15.jpg"> 图 15:肌肉传感器的会.这个数字显示了当肌肉传感器饱和时会发生什么;高原是当肌肉激活太强的电流增益设置的设备。<a href="http://cloudflare.jove.com/files/ftp_upload/55705/55705fig15large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table fo:keep-together.within-page="1"><tbody>
<tr>
<td></td>
			<td>MK I</td>
			<td>MK II</td>
		</tr>
<tr>
<td>肌电传感器</td>
			<td>通用生物传感器</td>
			<td>专用肌肉传感器</td>
		</tr>
<tr>
<td>无线</td>
			<td>单独的无线模块</td>
			<td>在微控制器板上</td>
		</tr>
<tr>
<td>无线的肌电信号？</td>
			<td>不</td>
			<td>是的</td>
		</tr>
<tr>
<td>电池</td>
			<td>9 V PP3</td>
			<td>150麻将脂</td>
		</tr>
<tr>
<td>操作时间</td>
			<td>1 h</td>
			<td>8 h</td>
		</tr>
<tr>
<td>生成时间</td>
			<td>5 h</td>
			<td>4 h</td>
		</tr>
<tr>
<td>总成本</td>
			<td>150美元</td>
			<td>80美元</td>
		</tr>
<tr>
<td>误报 (%)</td>
			<td>0</td>
			<td>0</td>
		</tr>
<tr>
<td>假阴性 (%)</td>
			<td>0</td>
			<td>4。7</td>
		</tr>
</tbody></table>表 1: 设备的比较。此表比较了设备的几个方面, 从设计到功能。
<table fo:keep-together.within-page="1"><tbody>
<tr>
<td></td>
			<td>固定 (最大)</td>
			<td>手移动 (最大)</td>
			<td>手指绷紧 (最小)</td>
			<td>阈值</td>
		</tr>
<tr>
<td>MK I</td>
			<td>25</td>
			<td>35</td>
			<td>215</td>
			<td>200</td>
		</tr>
<tr>
<td>MK II</td>
			<td>40</td>
			<td>280</td>
			<td>460</td>
			<td>400</td>
		</tr>
</tbody></table>表 2: 校准结果.该表显示的值, 同时保持手静止, 移动的手和手指紧张, 以及阈值选择。
<table fo:keep-together.within-page="1"><tbody>
<tr>
<td></td>
			<td colspan="3">假阴性数 (每三十年代测试一次)</td>
			<td colspan="2">误报数 (自发激活)</td>
		</tr>
<tr>
<td></td>
			<td>前45分钟</td>
			<td>45 min-1 h</td>
			<td>1-8 h</td>
			<td>第一 h</td>
			<td>1-8 h</td>
		</tr>
<tr>
<td>MK I</td>
			<td>0</td>
			<td>35</td>
			<td>N/A</td>
			<td>0</td>
			<td>N/A</td>
		</tr>
<tr>
<td>MK II</td>
			<td>4</td>
			<td>1</td>
			<td>40</td>
			<td>0</td>
			<td>0</td>
		</tr>
</tbody></table>表 3: 设备测试。两个设备之间的假阳性和假阴性的比较。
mk i 和 mk II 的补充代码文件: <a href="https://www.jove.com/files/ftp_upload/55705/BionicClicker.ino" target="_blank">请点击这里下载 "BionicClicker"</a> <a href="https://www.jove.com/files/ftp_upload/55705/BLEBionicClicker.ino" target="_blank">请点击这里下载 "BLEBionicClicker"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/BLEBoomTest.ino" target="_blank">请点击这里下载 "BLEBoomTest"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/BLEThresholdTest.ino" target="_blank">请点击这里下载 "BLEThresholdTest"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/BoomTest.ino" target="_blank">请点击这里下载 "BoomTest"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/ThresholdTest.ino" target="_blank">请点击这里下载 "ThresholdTest"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/Feather-Featherbase.stl" target="_blank">请点击这里下载 "羽毛 Featherbase. stl"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/Feather-Feathermid.stl" target="_blank">请点击这里下载 "羽毛 Feathermid. stl"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/Feather-Feathertop.stl" target="_blank">请点击这里下载 "羽毛 Feathertop. stl"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/Myo-Myobase.stl" target="_blank">请点击这里下载 "Myobase. stl"。</a> <a href="https://www.jove.com/files/ftp_upload/55705/Myo-Myolid.stl" target="_blank">请点击这里下载 "Myolid. stl"。</a>

Watch this Scientific Journal Video about The Bionic Clicker Mark I & II at JoVE.com

The Bionic Clicker Mark I & II

创建了一个设备, 以演示 electromyography-based 控制的位置受众。在最初的设备成功后, 第二个设备的功能更加灵活, 以供演示和研究之用。该协议描述了两种设备的生成和校准过程。

仿生响片 I &amp; II

In this manuscript, we present two 'Bionic Clicker' systems, the first designed to demonstrate electromyography (EMG) based control systems for ...

Research

JoVE Journal

Bioengineering

The Bionic Clicker Mark I &amp; II

16.3K 次观看.  University College London. 创建了一个设备, 以演示 electromyography-based 控制的位置受众。在最初的设备成功后, 第二个设备的功能更加灵活, 以供演示和研究之用。该协议描述了两种设备的生成和校准过程。

视频: The Bionic Clicker Mark I & II

High-resolution Fiber-optic Microendoscopy for in situ Cellular Imaging

Many biological and clinical studies require the longitudinal study and analysis of morphology and function with cellular level resolution. Traditionally, multiple experiments are run in parallel, with individual samples removed from the study at sequential time points for evaluation by light microscopy. Several intravital techniques have been developed, with confocal, multiphoton, and second harmonic microscopy all demonstrating their ability to be used for imaging in situ 1. With these systems, however, the required infrastructure is complex and expensive, involving scanning laser systems and complex light sources. Here we present a protocol for the design and assembly of a high-resolution microendoscope which can be built in a day using off-the-shelf components for under US$5,000. The platform offers flexibility in terms of image resolution, field-of-view, and operating wavelength, and we describe how these parameters can be easily modified to meet the specific needs of the end user.
We and others have explored the use of the high-resolution microendoscope (HRME) in in vitro cell culture 2-5, in excised 6 and living animal tissues 2,5, and in human tissues in vivo 2,7. Users have reported the use of several different fluorescent contrast agents, including proflavine 2-4, benzoporphyrin-derivative monoacid ring A (BPD-MA) 5, and fluoroscein 6,7, all of which have received full, or investigational approval from the FDA for use in human subjects. High-resolution microendoscopy, in the form described here, may appeal to a wide range of researchers working in the basic and clinical sciences. The technique offers an effective and economical approach which complements traditional benchtop microscopy, by enabling the user to perform high-resolution, longitudinal imaging in situ.

High-resolution Fiber-optic Microendoscopy for in situ Cellular Imaging

In many biological and clinical situations it is advantageous to study cellular processes as they evolve in their native microenvironment. Here we describe the assembly and use of a low-cost fiber-optic microscope which can provide real time imaging in cell culture, animal studies, and clinical patient studies.

高分辨率光纤内窥镜为原位细胞成像

Many biological and clinical studies require the longitudinal study and analysis of morphology and function with cellular level resolution.  ...

科研

生物工程

Organotypic Collagen I Assay: A Malleable Platform to Assess Cell Behaviour in a 3-Dimensional Context

Cell migration is fundamental to many aspects of biology, including development, wound healing, the cellular responses of the immune system, and metastasis of tumor cells. Migration has been studied on glass coverslips in order to make cellular dynamics amenable to investigation by light microscopy. However, it has become clear that many aspects of cell migration depend on features of the local environment including its elasticity, protein composition, and pore size, which are not faithfully represented by rigid two dimensional substrates such as glass and plastic 1. Furthermore, interaction with other cell types, including stromal fibroblasts 2 and immune cells 3, has been shown to play a critical role in promoting the invasion of cancer cells. Investigation at the molecular level has increasingly shown that molecular dynamics, including response to drug treatment, of identical cells are significantly different when compared in vitro and in vivo 4. 
Ideally, it would be best to study cell migration in its naturally occurring context in living organisms, however this is not always possible. Intermediate tissue culture systems, such as cell derived matrix, matrigel, organotypic culture (described here) tissue explants, organoids, and xenografts, are therefore important experimental intermediates. These systems approximate certain aspects of an in vivo environment but are more amenable to experimental manipulation such as use of stably transfected cell lines, drug treatment regimes, long term and high-resolution imaging. Such intermediate systems are especially useful as proving grounds to validate probes and establish parameters required to image the dynamic response of cells and fluorescent reporters prior to undertaking imaging in vivo 5. As such, they can serve an important role in reducing the need for experiments on living animals.

A method is described for the preparation of a 3-dimensional matrix consisting of collagen type I and primary human fibroblasts. This organotypic gel serves as a useful substrate to assess invasive cell migration because it mimics basic features of tissue stroma and is amenable to many forms of microscopy.

器官Ⅰ型胶原的含量可延展的平台，以评估细胞行为的一个3维的情况下

Cell migration is fundamental to many aspects of biology, including development, wound healing, the cellular responses of the immune system, and ...

Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface

The micropipette adhesion assay was developed in 1998 to measure two-dimensional (2D) receptor-ligand binding kinetics1. The assay uses a human red blood cell (RBC) as adhesion sensor and presenting cell for one of the interacting molecules. It employs micromanipulation to bring the RBC into contact with another cell that expresses the other interacting molecule with precisely controlled area and time to enable bond formation. The adhesion event is detected as RBC elongation upon pulling the two cells apart. By controlling the density of the ligands immobilized on the RBC surface, the probability of adhesion is kept in mid-range between 0 and 1. The adhesion probability is estimated from the frequency of adhesion events in a sequence of repeated contact cycles between the two cells for a given contact time. Varying the contact time generates a binding curve. Fitting a probabilistic model for receptor-ligand reaction kinetics1 to the binding curve returns the 2D affinity and off-rate.
The assay has been validated using interactions of Fc&gamma; receptors with IgG Fc1-6, selectins with glycoconjugate ligands6-9, integrins with ligands10-13, homotypical cadherin binding14, T cell receptor and coreceptor with peptide-major histocompatibility complexes15-19.
The method has been used to quantify regulations of 2D kinetics by biophysical factors, such as the membrane microtopology5, membrane anchor2, molecular orientation and length6, carrier stiffness9, curvature20, and impingement force20, as well as biochemical factors, such as modulators of the cytoskeleton and membrane microenvironment where the interacting molecules reside and the surface organization of these molecules15,17,19.
The method has also been used to study the concurrent binding of dual receptor-ligand species3,4, and trimolecular interactions19 using a modified model21.
The major advantage of the method is that it allows study of receptors in their native membrane environment. The results could be very different from those obtained using purified receptors17. It also allows study of the receptor-ligand interactions in a sub-second timescale with temporal resolution well beyond the typical biochemical methods.
To illustrate the micropipette adhesion frequency method, we show kinetics measurement of intercellular adhesion molecule 1 (ICAM-1) functionalized on RBCs binding to integrin &alpha;L&beta;2 on neutrophils with dimeric E-selectin in the solution to activate &alpha;L&beta;2.

Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface

An adhesion frequency assay for measuring receptor-ligand interaction kinetics when both molecules are anchored on the surfaces of the interacting cells is described. This mechanically-based assay is exemplified using a micropipette-pressurized human red blood cell as adhesion sensor and integrin &alpha;L&beta;2 and intercellular adhesion molecule-1 as interacting receptors and ligands.

粘附频率含量为原位跨交界的分子相互作用，在细胞与细胞界面的动力学分析

The micropipette adhesion assay was developed in 1998 to measure two-dimensional (2D) receptor-ligand binding kinetics1. The assay uses a human red blood ...

Silk Film Culture System for in vitro Analysis and Biomaterial Design

Silk films are promising protein-based biomaterials that can be fabricated with high fidelity and economically within a research laboratory environment 1,2 . These materials are desirable because they possess highly controllable dimensional and material characteristics, are biocompatible and promote cell adhesion, can be modified through topographic patterning or by chemically altering the surface, and can be used as a depot for biologically active molecules for drug delivery related applications 3-8 . In addition, silk films are relatively straightforward to custom design, can be designed to dissolve within minutes or degrade over years in vitro or in vivo, and are produce with the added benefit of being transparent in nature and therefore highly suitable for imaging applications 9-13. The culture system methodology presented here represents a scalable approach for rapid assessments of cell-silk film surface interactions. Of particular interest is the use of surface patterned silk films to study differences in cell proliferation and responses of cells for alignment 12,14 . The seeded cultures were cultured on both micro-patterned and flat silk film substrates, and then assessed through time-lapse phase-contrast imaging, scanning electron microscopy, and biochemical assessment of metabolic activity and nucleic acid content. In summary, the silk film in vitro culture system offers a customizable experimental setup suitable to the study of cell-surface interactions on a biomaterial substrate, which can then be optimized and then translated to in vivo models. Observations using the culture system presented here are currently being used to aid in applications ranging from basic cell interactions to medical device design, and thus are relevant to a broad range of biomedical fields.

Silk Film Culture System for in vitro Analysis and Biomaterial Design

Silk films are a novel class of biomaterials readily customizable for an array of biomedical applications. The presented silk film culture system is highly adaptable to a variety of in vitro analyses. This system represents a biomaterial design platform offering in vitro optimization before direct translation to in vivo models.

丝绸电影文化系统在体外分析和生物材料设计

Silk films are promising protein-based biomaterials that can be fabricated with high fidelity and economically within a research laboratory environment ...

Cell Patterning on Photolithographically Defined 
Parylene-C: SiO2 Substrates

Cell patterning platforms support broad research goals, such as construction of predefined in vitro neuronal networks and the exploration of certain central aspects of cellular physiology. To easily combine cell patterning with Multi-Electrode Arrays (MEAs) and silicon-based &#8216;lab on a chip&#8217; technologies, a microfabrication-compatible protocol is required. We describe a method that utilizes deposition of the polymer parylene-C on SiO2&#160;wafers. Photolithography enables accurate and reliable patterning of parylene-C at micron-level resolution. Subsequent activation by immersion in fetal bovine serum (or another specific activation solution) results in a substrate in which cultured cells adhere to, or are repulsed by, parylene or SiO2 regions respectively. This technique has allowed patterning of a broad range of cell types (including primary murine hippocampal cells, HEK 293 cell line, human neuron-like teratocarcinoma cell line, primary murine cerebellar granule cells, and primary human glioma-derived stem-like cells). Interestingly, however, the platform is not universal; reflecting the importance of cell-specific adhesion molecules. This cell patterning process is cost effective, reliable, and importantly can be incorporated into standard microfabrication (chip manufacturing) protocols, paving the way for integration of microelectronic technology.

Cell Patterning on Photolithographically Defined 
Parylene-C: SiO2 Substrates

This protocol describes a microfabrication-compatible method for cell patterning on SiO2. A predefined parylene-C design is photolithographically printed on SiO2 wafers. Following incubation with serum (or other activation solution) cells adhere specifically to (and grow according to the conformity of) underlying parylene-C, whilst being repulsed by SiO2 regions.

在光刻技术定义的聚对二甲苯-C细胞图案:二氧化硅<sub&gt; 2</sub&gt;基板

Cell patterning platforms support broad research goals, such as construction of predefined in vitro neuronal networks and the exploration of certain ...

An Improved Method for the Preparation of Type I Collagen From Skin

Soluble type 1 collagen (COL1) is used extensively as an adhesive substrate for cell cultures and as a cellular scaffold for regenerative applications. Clinically, this protein is widely used for cosmetic surgery, dermal injections, bone grafting, and reconstructive surgery. The sources of COL1 for these procedures are commonly nonhuman, which increases the potential for inflammation and rejection as well as xenobiotic disease transmission. In view of this, a method to efficiently and quickly purify COL1 from limited quantities of autologously-derived tissues would circumvent many of these issues; however, standard isolation protocols are lengthy and often require large quantities of collagenous tissues. Here, we demonstrate an efficient COL1 extraction method that reduces the time needed to isolate and purify this protein from about 10 days to less than 3 hr. We chose the dermis as our tissue source because of its availability during many surgical procedures. This method uses traditional extraction buffers combined with forceful agitation and centrifugal filtration to obtain highly-pure, soluble COL1 from small amounts of corium. Briefly, dermal biopsies are washed thoroughly in ice-cold dH2O after removing fat, connective tissue, and hair. The skin samples are stripped of noncollagenous proteins and polysaccharides using 0.5 M sodium acetate and a high speed bench-top homogenizer. Collagen from residual solids is subsequently extracted with a 0.075 M sodium citrate buffer using the homogenizer. These extracts are purified using 100,000 MW cut-off centrifugal filters that yield COL1 preparations of comparable or superior quality to commercial products or those obtained using traditional procedures. We anticipate this method will facilitate the utilization of autologously-derived COL1 for a multitude of research and clinical applications.

Traditional procedures for the isolation of soluble type 1 collagen (COL1) require about 10 days from start to finish because of lengthy buffer incubations and laborious resuspensions of fibrils. Here, we describe a means to purify COL1 from small dermal biopsies in less than 3 hr.

对于I型胶原蛋白的制备皮肤方法的改进

Soluble type 1 collagen (COL1) is used extensively as an adhesive substrate for cell cultures and as a cellular scaffold for regenerative applications. ...

In vivo and In vitro Rearing of Entomopathogenic Nematodes (Steinernematidae and Heterorhabditidae)

Entomopathogenic nematodes (EPN) (Steinernematidae and Heterorhabditidae) have a mutualistic partnership with Gram-negative Gamma-Proteobacteria in the family Enterobacteriaceae. Xenorhabdus bacteria are associated with steinernematids nematodes while Photorhabdus are symbionts of heterorhabditids. Together nematodes and bacteria form a potent insecticidal complex that kills a wide range of insect species in an intimate and specific partnership. Herein, we demonstrate in vivo and in vitro techniques commonly used in the rearing of these nematodes under laboratory conditions. Furthermore, these techniques represent key steps for the successful establishment of EPN cultures and also form the basis for other bioassays that utilize these organisms for research. The production of aposymbiotic (symbiont&#8211;free) nematodes is often critical for an in-depth and multifaceted approach to the study of symbiosis. This protocol does not require the addition of antibiotics and can be accomplished in a short amount of time with standard laboratory equipment. Nematodes produced in this manner are relatively robust, although their survivorship in storage may vary depending on the species used. The techniques detailed in this presentation correspond to those described by various authors and refined by P. Stock&#8217;s Laboratory, University of Arizona (Tucson, AZ, USA). These techniques are distinct from the body of techniques that are used in the mass production of these organisms for pest management purposes.

In vivo and In vitro Rearing of Entomopathogenic Nematodes Steinernematidae and Heterorhabditidae

The goal of this presentation is to demonstrate in vivo and in vitro techniques for the rearing of entomopathogenic nematodes. In vivo methods consider the rearing of these nematodes with an insect host, whereas the in vitro methods utilize rich agar media.

在体内和昆虫病原线虫体外培育（ 斯氏和异小 ）

Entomopathogenic nematodes (EPN) (Steinernematidae and Heterorhabditidae) have a mutualistic partnership with Gram-negative Gamma-Proteobacteria in the ...

Delivery of Therapeutic siRNA to the CNS Using Cationic and Anionic Liposomes

Prion diseases result from the misfolding of the normal, cellular prion protein (PrPC) to an abnormal protease resistant isomer called PrPRes. The emergence of prion diseases in wildlife populations and their increasing threat to human health has led to increased efforts to find a treatment for these diseases. Recent studies have found numerous anti-prion compounds that can either inhibit the infectious PrPRes isomer or down regulate the normal cellular prion protein. However, most of these compounds do not cross the blood brain barrier to effectively inhibit PrPRes formation in brain tissue, do not specifically target neuronal PrPC, and are often too toxic to use in animal or human subjects. 
We investigated whether siRNA delivered intravascularly and targeted towards neuronal PrPC is a safer and more effective anti-prion compound. This report outlines a protocol to produce two siRNA liposomal delivery vehicles, and to package and deliver PrP siRNA to neuronal cells. The two liposomal delivery vehicles are 1) complexed-siRNA liposome formulation using cationic liposomes (LSPCs), and 2) encapsulated-siRNA liposome formulation using cationic or anionic liposomes (PALETS). For the LSPCs, negatively charged siRNA is electrostatically bound to the cationic liposome. A positively charged peptide (RVG-9r [rabies virus glycoprotein]) is added to the complex, which specifically targets the liposome-siRNA-peptide complexes (LSPCs) across the blood brain barrier (BBB) to acetylcholine expressing neurons in the central nervous system (CNS). For the PALETS (peptide addressed liposome encapsulated therapeutic siRNA), the cationic and anionic lipids were rehydrated by the PrP siRNA. This procedure results in encapsulation of the siRNA within the cationic or anionic liposomes. Again, the RVG-9r neuropeptide was bound to the liposomes to target the siRNA/liposome complexes to the CNS. Using these formulations, we have successfully delivered PrP siRNA to AchR-expressing neurons, and decreased the PrPC expression of neurons in the CNS.

The goal of this protocol is to use cationic/anionic liposomes with a neuro-targeting peptide as a CNS delivery system to enable siRNA to cross the BBB. The optimization of a delivery system for treatments, like siRNA, would allow for more treatment options for prion and other neurodegenerative diseases.

治疗的siRNA递送至CNS使用阳离子和阴离子脂质体

Prion diseases result from the misfolding of the normal, cellular prion protein (PrPC) to an abnormal protease resistant isomer called PrPRes.  The ...

Determination of Zeta Potential via Nanoparticle Translocation Velocities through a Tunable Nanopore: Using DNA-modified Particles as an Example

Nanopore technologies, known collectively as Resistive Pulse Sensors (RPS), are being used to detect, quantify and characterize proteins, molecules and nanoparticles. Tunable resistive pulse sensing (TRPS) is a relatively recent adaptation to RPS that incorporates a tunable pore that can be altered in real time. Here, we use TRPS to monitor the translocation times of DNA-modified nanoparticles as they traverse the tunable pore membrane as a function of DNA concentration and structure (i.e., single-stranded to double-stranded DNA).
TRPS is based on two Ag/AgCl electrodes, separated by an elastomeric pore membrane that establishes a stable ionic current upon an applied electric field. Unlike various optical-based particle characterization technologies, TRPS can characterize individual particles amongst a sample population, allowing for multimodal samples to be analyzed with ease. Here, we demonstrate zeta potential measurements via particle translocation velocities of known standards and apply these to sample analyte translocation times, thus resulting in measuring the zeta potential of those analytes.
As well as acquiring mean zeta potential values, the samples are all measured using a particle-by-particle perspective exhibiting more information on a given sample through sample population distributions, for example. Of such, this method demonstrates potential within sensing applications for both medical and environmental fields.

Here we use a polyurethane tunable nanopore integrated into a resistive pulse sensing technique to characterize nanoparticles surface chemistry via the measurement of particle translocation velocities, which can be used to determine the zeta potential of individual nanoparticles.

通过通过纳米孔可调纳米易位流速Zeta电位测定:利用DNA修饰颗粒为例

Nanopore technologies, known collectively as Resistive Pulse Sensors (RPS), are being used to detect, quantify and characterize proteins, molecules and ...

The Organoid Reconstitution Assay (ORA) for the Functional Analysis of Intestinal Stem and Niche Cells

The intestinal epithelium is characterized by an extremely rapid turnover rate. In mammals, the entire epithelial lining is renewed within 4 - 5 days. Adult intestinal stem cells reside at the bottom of the crypts of Lieberk&#252;hn, are earmarked by expression of the Lgr5 gene, and preserve homeostasis through their characteristic high proliferative rate1. Throughout the small intestine, Lgr5+ stem cells are intermingled with specialized secretory cells called Paneth cells. Paneth cells secrete antibacterial compounds (i.e., lysozyme and cryptdins/defensins) and exert a controlling role on the intestinal flora. More recently, a novel function has been discovered for Paneth cells, namely their capacity to provide niche support to Lgr5+ stem cells through several key ligands as Wnt3, EGF, and Dll12.
When isolated ex vivo and cultured in the presence of specific growth factors and extracellular matrix components, whole intestinal crypts give rise to long-lived and self-renewing 3D structures called organoids that highly resemble the crypt-villus epithelial architecture of the adult small intestine3. Organoid cultures, when established from whole crypts, allow the study of self-renewal and differentiation of the intestinal stem cell niche, though without addressing the contribution of its individual components, namely the Lgr5+ and Paneth cells.
Here, we describe a novel approach to the organoid assay that takes advantage of the ability of Paneth and Lgr5+ cells to associate and form organoids when co-cultured. This approach, here referred to as &#34;organoid reconstitution assay&#34; (ORA), allows the genetic and biochemical modification of Paneth or Lgr5+ stem cells, followed by reconstitution into organoids. As such, it allows the functional analysis of the two main components of the intestinal stem cell niche.

The Organoid Reconstitution Assay ORA for the Functional Analysis of Intestinal Stem and Niche Cells

Intestinal organoid cultures are established from whole crypts and do not allow the analysis of self-renewal and differentiation in a cell-specific fashion. This protocol describes reconstitution of sorted stem (Lgr5+) and niche (Paneth) cells, which give rise to organoids while enabling their prior biochemical and genetic modification and functional analysis.

化重构法 (的) 对肠道干细胞和小生境细胞功能分析的研究

The intestinal epithelium is characterized by an extremely rapid turnover rate. In mammals, the entire epithelial lining is renewed within 4 - 5 days. ...