Name: fabrication of microscope stage for vertical observation with temperature control function
Uploaded: 2019-07-31T14:00:00
Description: Watch this Scientific Journal Video about Fabrication of Microscope Stage for Vertical Observation with Temperature Control Function at JoVE.com

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1. 设计
<ol><li>铝板的制造<ol>
<li>在尺寸为 150 mm x 200 mm x 2 mm 的铝板中心切割 101 mm 孔,用激光加工机用作最前沿板。机器爪子在八点,以贴上两个橡皮筋跨越的长度,或两个横跨这个板的宽度(参见补充图1A和补充图2A)。</li>
		<li>在另一块 150 mm x 200 mm x 5 mm 铝板的中心切割一个 130 mm 孔,用激光加工机用作中上板。机器八个凹槽,用于在长度的两个点上连接橡皮筋,或两个穿过此板...

<ol>
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运动硅藻细胞的轨迹分析是评估硅藻动性的有用方法。然而,虽然正常的倒置显微镜水平观察样品,但它不适合观察重力或垂直方向的浮动运动的影响。这里开发和描述的是一个垂直显微镜阶段,具有温度控制,并连接到倒置显微镜,已旋转90°。这种具有温度控制的显微镜阶段允许观察硅藻细胞的温度相关垂直运动。
协议中的一个关键步骤是控制器电路设计。为保证安全,采用了断?...

光学显微镜是一种通过放大镜头和可见光的样品来增加可观察细节的技术。在光学显微镜中,光被定向到样品上,然后通过放大镜捕捉透镜进行反射、反射或荧光灯的捕获,以便观察。有多种显微镜在设计上有所不同,以适应不同的用途和观察方法。不同的设计包括立式显微镜,其结构是从下方照亮样品,以便从上面观察,以及倒置显微镜,从上面照亮样品,以便从下方观察。直立显微镜是最常见和广泛使用的设计。倒置显微镜通常用于观察不能让透镜与上方距离接近的样品,例如附着在容器底部的培养细胞。许多研究小组使用倒置显微镜1、2、3、4、5、6、7报告在广泛领域的观测结果。还开发了许多额外的设备,利用倒置显微镜8,9,10,11,12,13的功能.
目前,在所有传统的显微镜设计中,显微镜级是水平的,因此不适合观察在垂直平面上产生的样品运动(由于重力、浮力、运动等)。为了使这些观察成为可能,显微镜级和光路径必须旋转到垂直。垂直舞台需要垂直安装玻璃滑梯或样品容器,如培养皿到舞台上。为了解决这个问题,已经设计了一个侧向倒置显微镜,倾斜90°。但是,用胶带或其他粘合剂连接样品不会产生必要的长期静止状态。此处描述的是一种可实现必要稳定性的设备。此器件允许在垂直平面中观察样本随时间移动的时间。硅橡胶加热器的安装也使得观察温度变化对样品行为的影响成为可能。温度数据通过 Wi-Fi 传输到互联网服务器,温度设置和日志监控可以从 PC 或智能手机远程控制。据我们所知,在以前的研究中尚未报告附着在倾斜 90° 的侧向倾斜显微镜上的阶段。
显微镜阶段由三个铝板组成。中间铝板安装在连接到舞台的下部铝板上。含有温度传感器的硅橡胶连接在中铝板和上部铝板之间。橡皮筋用于粘贴样品。爪子附在上铝板的左右四个点,以固定橡胶带。温度调节器的控制电路接收嵌入硅橡胶的温度传感器的信号,并通过脉冲宽度调制(PWM)方法调节电能。温度可以1°C为增量逐渐升高至50°C。此器件适用于垂直采样运动可能依赖于温度的应用。
本报告提供了温度对硅藻浮现象的影响示例。作为硅藻观测研究的例子,细胞簇的沉积速度测量、运动分析、超细结构研究等已报告14、15、16、17,18,19,20,21,22,23.与光合生物一起漂浮在水中的硅藻的具体重力略高于水,因此容易下沉;然而,如果发生轻微的对流,它们也会上升。为了研究这一现象,玻璃滑动垂直地贴在显微镜的舞台上,并观察到温度升高对硅藻垂直运动的影响。

<table>
	<tbody>
		<tr>
			<td>AC adapter 12V2A</td>
			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
			<td>AD-D120P200</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>ADS1015 Substrate</td>
			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
			<td>adafruit PRODUCT ID: 1083</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Alminium Plate (Back Side Plate)</td>
			<td>Inoval Co., Ltd.</td>
			<td>W 150mm&#215;L 200&#13212;&#215;T 1.5mm</td>
			<td>Gifu, Japan</td>
		</tr>
		<tr>
			<td>Alminium Plate (Forefront Plate)</td>
			<td>Inoval Co., Ltd.</td>
			<td>W 150mm&#215;L 200&#13212;&#215;T 2mm</td>
			<td>Gifu, Japan</td>
		</tr>
		<tr>
			<td>Alminium Plate (Middle Lower Plate)</td>
			<td>Inoval Co., Ltd.</td>
			<td>W 150mm&#215;L 200&#13212;&#215;T 4mm</td>
			<td>Gifu, Japan</td>
		</tr>
		<tr>
			<td>Alminium Plate (Middle Upper Plate)</td>
			<td>Inoval Co., Ltd.</td>
			<td>W 150mm&#215;L 200&#13212;&#215;T 5mm</td>
			<td>Gifu, Japan</td>
		</tr>
		<tr>
			<td>Aluminum Pedestal (Lower Plate)</td>
			<td>Inoval Co., Ltd.</td>
			<td>D 100mm&#215;T 3mm (30&#934;)</td>
			<td>Gifu, Japan</td>
		</tr>
		<tr>
			<td>Aluminum Pedestal (Upper Plate)</td>
			<td>Inoval Co., Ltd.</td>
			<td>D 100mm&#215;T 3mm (30&#934;)</td>
			<td>Gifu, Japan</td>
		</tr>
		<tr>
			<td>Bold Modified Basal Freshwater Nutrient Solution</td>
			<td>Sigma-Aldrich Co. LLC</td>
			<td>B5282-500ML</td>
			<td>St. Louis, USA</td>
		</tr>
		<tr>
			<td>Controller Case</td>
			<td>Marutsu Elec Co., Ltd.</td>
			<td>pff-13-3-9</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>CPU</td>
			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
			<td>ESP-WROOM-02D</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Inverted microscope</td>
			<td>Olympus Corporation</td>
			<td>CKX 53</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Low temperature hardening epoxy resin adhesive</td>
			<td>ThreeBond Co., Ltd.</td>
			<td>TB2086M</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Multi-turn semi-fixed volume Vertical type 500 &#937;</td>
			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
			<td>3296W-1-501LF</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>OLED module</td>
			<td>Akihabara Inc.</td>
			<td>M096P4W</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Pressed Cork (For supporting electrode )</td>
			<td>Tera Co., Ltd.</td>
			<td>W 42mm&#215;L 30&#13212;</td>
			<td>Ishikawa, Japan</td>
		</tr>
		<tr>
			<td>Pressed Cork (Lower Disk)</td>
			<td>Tera Co., Ltd.</td>
			<td>D 100mm&#215;T 0.5mm (20&#934;)</td>
			<td>Ishikawa, Japan</td>
		</tr>
		<tr>
			<td>Pressed Cork (Upper Disk)</td>
			<td>Tera Co., Ltd.</td>
			<td>D 100mm&#215;T 2.5mm (20&#934;)</td>
			<td>Ishikawa, Japan</td>
		</tr>
		<tr>
			<td>Rotary encoder with switch with 2 color LED</td>
			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
			<td>P-05772</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Silicone rubber heater</td>
			<td>Three High Co., Ltd.</td>
			<td>D 100mm&#215;T 2.5mm (20&#934;)</td>
			<td>Kanagawa, Japan</td>
		</tr>
		<tr>
			<td>Substrate</td>
			<td>Seeed Technology Co., Ltd.</td>
			<td>mh5.0</td>
			<td>Shenzhen, China</td>
		</tr>
		<tr>
			<td>Temperature sensor</td>
			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
			<td>NXFT15XH103FA2B050</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Three-terminal DC / DC regulator 3.3 V</td>
			<td>Marutsu Elec Co., Ltd.</td>
			<td>BR301</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Universal Flexible Arm</td>
			<td>Banggood Technology Co., Ltd.</td>
			<td>YP-003-2</td>
			<td>Hong Kong, China</td>
		</tr>
		<tr>
			<td>USB cable USB-A - MicroUSB</td>
			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
			<td>USB CABLE A-MICROB</td>
			<td>Tokyo, Japan</td>
		</tr>
		<tr>
			<td>Video Canera</td>
			<td>Sony Corporation</td>
			<td>HDR-CX590</td>
			<td>Tokyo, Japan</td>
		</tr>
	</tbody>
</table>

fabrication of microscope stage for vertical observation with temperature control function

样品通常放在水平显微镜台上进行微观观察。然而,为了观察重力对样品的影响或研究漂浮行为,有必要使显微镜阶段垂直。为此,设计了一个侧向倒置显微镜,倾斜90°。要用显微镜观察样品,必须将培养皿或玻璃玻片等样品容器垂直固定到舞台上。开发了一种在垂直显微镜舞台上将样品容器固定到位的装置,并在此进行了描述。此器件连接到该级允许观察垂直平面中的样品动力学。使用硅橡胶加热器调节温度的能力还允许观察温度相关样品行为。此外,温度数据传输到互联网服务器。温度设置和日志监控可以从 PC 或智能手机远程控制。

图 2显示了橡胶加热器的温度分布。橡胶加热器的表面温度在每个温度下都是均匀的。图 3显示了测量温度对设置温度变化的响应能力。橙色线显示设定的温度,蓝线显示样品温度的变化。测得的值对设置更改的过冲很小,跟踪速度很快。
观察到硅藻细胞,以提供使用此设备的特定示例?...

Watch this Scientific Journal Video about Fabrication of Microscope Stage for Vertical Observation with Temperature Control Function at JoVE.com

Fabrication of Microscope Stage for Vertical Observation with Temperature Control Function

具有温度控制功能的垂直观测显微镜级的制造

这里介绍的是一个使用温度控制的显微镜级的协议,它允许将样品容器安装在垂直显微镜上。

Samples are usually placed onto a horizontal microscope stage for microscopic observation. However, to observe the influence of gravity on a sample or ...

该器件可用于将样品固定到垂直显微镜阶段，以便观察重力的影响，并处理感兴趣的样品。将本设备连接到阶段可观察垂直框架中的样品动力学，并允许观察与温度相关的样品行为。要制造铝板，请使用激光加工机在 150 由 200 比 200 的铝板中心切割一个 101 毫米的孔，以用作最前列板。在板外侧八个点处制作爪子，使两个橡皮筋贴在板的长度或宽度上。接下来在150%200的铝板中心切一个130毫米的孔，用作中上板，并制作8个凹口，使两个橡皮筋在板的长度或宽度上放置。然后在 150 % 200 的铝板中心切割一个 130 毫米的孔，用作中下板，在 150 到 200 的铝板的中心切割一个 30 毫米孔，用作底板。要制造基座，在100毫米直径、3毫米厚的铝板中心切一个30毫米的孔，并在板的一侧做一个42毫米宽、30毫米深的凹口。然后在直径100毫米、4毫米厚的铝板的中心切一个30毫米的孔，从板的中心钻三个三毫米孔，25毫米，彼此之间120度。为制造压榨软木盘，使用喷水切割机在100毫米直径、2毫米厚的压塞软盘的中心切一个20毫米的孔，并在盘的每一侧切割一个42毫米宽、30毫米深、一个4毫米宽、5毫米深的圆孔。接下来在100毫米直径、1毫米厚的压软木盘的中心切一个20毫米的孔，在圆盘的每一侧做一个42毫米宽、30毫米深、一个4毫米宽、40毫米深的凹槽。然后从直径为 100 毫米的圆盘上切割一个 42 毫米宽、30 毫米深的压紧软木板。要制造硅橡胶加热器，在直径为 100 毫米、2.5 毫米厚的硅橡胶盘的中心切一个 20 毫米的孔，并配有内置的镍丝。然后按演示将制造部件堆叠起来，并按要求用螺钉或粘合剂固定适当的部件。使用专用电缆将橡胶加热器和控制器外壳的加热器合并，将显微镜级连接到系统，并使用控制器装备 Wi-Fi 信号并控制橡胶加热器的电流。构建系统后，将热敏电阻导线连接到控制器外壳上的传感器端子，并接收热敏电阻测量的温度信号。使用控制器上的旋钮更改设定温度。然后通过互联网将测量温度、设定温度和测量时间信息从控制器传输到服务器。要分析样品，请将样品放在垂直于地面的显微镜舞台上，并使用四个长爪用两个橡皮筋固定样品。使用控制器将温度设置为 40 摄氏度，并检查显示屏上的温度。然后按旋钮开始温度控制。蓝色 LED 指示灯将点亮，指示热源启动。在这些数字中，显示了橡胶加热器的代表性温度分布。橡胶加热器的表面温度在每个温度下均匀。此处显示了测量温度对设置温度变化的响应能力的示例。橙色线指示设定温度，蓝线表示样品温度的变化。如果设备组装正确，则测量值对设置更改的过冲通常较小，并且跟踪速度很快。在本次实验中，成功记录了硅藻的温度依赖性垂直运动，从而可以检测出硅藻垂直运动的位点。然后，可以通过直接观测来可视化热对流对硅藻细胞垂直漂浮现象的影响。当传感器与样品断开时，或者微控制器不能正常工作时，检查加热器的电流是否与微控制器断开。使用这种方法，可以观察到温度变化对生物体在水中的垂直运动的影响。由于附着在显微镜上的舞台部分的结构复杂，未来的研究将涉及该结构的简化问题。要将样品冷却到低于室温，需要复杂的冷却装置，这也是未来工作考虑的问题。

Research

JoVE Journal

Engineering

Fabrication of Gate-tunable Graphene Devices for Scanning Tunneling Microscopy Studies with Coulomb Impurities

对于扫描隧道显微学与库仑杂质制造门可调谐石墨烯器件

Owing to its relativistic low-energy charge carriers, the interaction between graphene and various impurities leads to a wealth of new physics and degrees ...

科研

工程学

Fabrication of Low Temperature Carbon Nanotube Vertical Interconnects Compatible with Semiconductor Technology

低温碳纳米管垂直互连的制作兼容半导体技术

We demonstrate a method for the low temperature growth (350 &#176;C) of vertically-aligned carbon nanotubes (CNT) bundles on electrically conductive ...

A Fabrication Method for Highly Stretchable Conductors with Silver Nanowires

对于高伸缩的导线与银纳米线制作方法

Stretchable electronics are identified as a key technology for electronic applications in the next generation. One of the challenges in fabrication of ...

Free-form Light Actuators &#8212; Fabrication and Control of Actuation in Microscopic Scale

Free-form Light Actuators ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Â Fabrication and Control of Actuation in Microscopic Scale

自由形式的光驱动器 - 制作和开动控制在微观尺度

Liquid crystalline elastomers (LCEs) are smart materials capable of reversible shape-change in response to external stimuli, and have attracted ...

High Temperature Fabrication of Nanostructured Yttria-Stabilized-Zirconia (YSZ) Scaffolds by In Situ Carbon Templating Xerogels

High Temperature Fabrication of Nanostructured Yttria-Stabilized-Zirconia YSZ Scaffolds by In Situ Carbon Templating Xerogels

通过纳米氧化钇稳定的二氧化锆（YSZ）的支架的高温加工<em&gt;原位</em&gt;碳模板干凝胶

We demonstrate a method for the high temperature fabrication of porous, nanostructured yttria-stabilized-zirconia (YSZ, 8 mol% yttria - 92 mol% zirconia) ...

Preparation of Large-area Vertical 2D Crystal Hetero-structures Through the Sulfurization of Transition Metal Films for Device Fabrication

硫化过渡金属膜制备大面积垂直2D 晶体杂化结构

We have demonstrated that through the sulfurization of transition metal films such as molybdenum (Mo) and tungsten (W), large-area and uniform transition ...

Radio Frequency Magnetron Sputtering of GdBa2Cu3O7&#8722;&#948;/ La0.67Sr0.33MnO3 Quasi-bilayer Films on SrTiO3 (STO) Single-crystal Substrates

Radio Frequency Magnetron Sputtering of GdBa2Cu3O7ÃƒÆ’Ã‚Â¢Ãƒâ€¹Ã¢â‚¬Â 'ÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â´/ La0.67Sr0.33MnO3 Quasi-bilayer Films on SrTiO3 STO Single-crystal Substrates

Gdba2cu 3 o7 的射频磁控管散射-/la0.67sr 0.67 mno 3 准双层薄膜上的 srtio 3 (sto) 单晶基板上

Here, we demonstrate a method of coating ferromagnetic La0.67Sr0.33MnO3 (LSMO) nanoparticles on (001) SrTiO3 (STO) single-crystal substrates by radio ...

Single-Digit Nanometer Electron-Beam Lithography with an Aberration-Corrected Scanning Transmission Electron Microscope

用畸变校正扫描透射电镜进行单数字纳米电子束光刻

We demonstrate extension of electron-beam lithography using conventional resists and pattern transfer processes to single-digit nanometer dimensions by ...

Fabrication of Soft Pneumatic Network Actuators with Oblique Chambers

斜腔软气动网络驱动器的研制

Soft pneumatic network actuators have become one of the most promising actuation devices in soft robotics which benefits from their large bending ...

Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering

基于刺激拉曼散射的非线性显微镜的实现

Stimulated Raman scattering (SRS) microscopy uses near-infrared excitation light; therefore, it shares many multi-photon microscopic imaging properties. ...