The authors have no acknowledgements.

1. Design
<ol>
	<li>Fabrication of aluminum plates
	<ol>
		<li>Cut a 101 mm hole in the center of an aluminum plate of dimensions 150 mm x 200 mm x 2 mm to be used as the forefront plate with a laser processing machine. Machine claws at eight points to affix two rubber bands across the length, or two across the width of this plate (see Supplemental Figure 1A and Supplemental Figure 2A).</li>
		<li>Cut a 130 mm hole in the center of another 150 mm x 200 mm x 5 mm aluminum plate to be used as the middle upper plate with a laser processing machine. Machine eight notches for attaching rubber bands at two points across the length, or two across the width of this plate (see Supplemental Figure 1B and Supplemental Figure 2B).</li>
		<li>Cut a 130 mm hole in the center of a 150 mm x 200 mm x 4 mm aluminum plate to be used as the middle lower plate with a laser processing machine (see Supplemental Figure 1C and Supplemental Figure 2C).</li>
		<li>Cut a 30 mm hole in the center of a 150 mm x 200 mm x 1.5 mm aluminum plate to be used as the base plate (see Supplemental Figure 1D and Supplemental Figure 2D).</li>
	</ol>
	</li>
	<li>Fabrication of two aluminum pedestal
	<ol>
		<li>Cut a 30 mm hole in the center of the aluminum plate (100 mm diameter, 3 mm thickness) and make a notch from one side with the dimensions 42 mm wide x 30 mm deep (see Supplemental Figure 3A).</li>
		<li>Cut a 30 mm hole in the center of the plate in an aluminum plate (100 mm diameter, 4 mm thickness) and drill three 3 mm holes located 25 mm from the center, spaced 120&#176; from each other (see Supplemental Figure 3B).</li>
	</ol>
	</li>
	<li>Fabrication of three pressed cork disc
	<ol>
		<li>Cut a 20 mm hole in the center of the pressed cork disc (100 mm diameter, 2 mm thickness) with a water jet cutting machine. Make one cut 42 mm across x 30 mm deep, then one cut 4 mm wide x 5 mm deep (see Supplemental Figure 4A).</li>
		<li>Cut a 20 mm hole in the center of the pressed cork disc of dimensions 100 mm diameter, 1 mm thickness with a water jet cutting machine. Make a cut 42 mm across x 30 mm deep, a cut 4 mm wide x 40 mm deep (see Supplemental Figure 4B).</li>
		<li>Cut a pressed cork plate from a 100 mm diameter disc with a 42 mm width and 30 mm depth. Two sheets of 1 mm thickness and one sheet of 2 mm thickness are required (see Supplemental Figure 4C).</li>
	</ol>
	</li>
	<li>Fabrication of silicone rubber heater
	<ol>
		<li>Fabricate a heater using a 100 mm diameter disc of 2.5 mm thick silicon rubber with built-in Nichrome wire and cut a 20 mm hole in the center of the disc (see Supplemental Figure 5).</li>
	</ol>
	</li>
	<li>Assemble parts described in steps 1.1&#8211;1.4 by stacking them as shown in Supplemental Figure 6.</li>
	<li>To construct a microscope stage, refer to Supplemental Figure 6, cross-section of the microscope stage. Fix <img alt="Equation 1" src="/files/ftp_upload/59799/59799eq1.jpg" /> and <img alt="Equation 2" src="/files/ftp_upload/59799/59799eq2.jpg" />, then <img alt="Equation 3" src="/files/ftp_upload/59799/59799eq3.jpg" /> and <img alt="Equation 4" src="/files/ftp_upload/59799/59799eq4.jpg" /> with screws. Fix <img alt="Equation 4" src="/files/ftp_upload/59799/59799eq4.jpg" /> and <img alt="Equation 5" src="/files/ftp_upload/59799/59799eq5.jpg" /> with screws. Fix <img alt="Equation 2" src="/files/ftp_upload/59799/59799eq2.jpg" /> and <img alt="Equation 3" src="/files/ftp_upload/59799/59799eq3.jpg" />, <img alt="Equation 6" src="/files/ftp_upload/59799/59799eq6.jpg" /> and <img alt="Equation 5" src="/files/ftp_upload/59799/59799eq5.jpg" />, <img alt="Equation 6" src="/files/ftp_upload/59799/59799eq6.jpg" /> and <img alt="Equation 7" src="/files/ftp_upload/59799/59799eq7.jpg" />, <img alt="Equation 7" src="/files/ftp_upload/59799/59799eq7.jpg" /> and <img alt="Equation 8" src="/files/ftp_upload/59799/59799eq8.jpg" />, and <img alt="Equation 5" src="/files/ftp_upload/59799/59799eq5.jpg" /> and <img alt="Equation 9" src="/files/ftp_upload/59799/59799eq9.jpg" /> with adhesive.</li>
</ol>
2. Hardware design outlines
<ol>
	<li>Prepare a &#8220;power supply and programming circuit&#8221; as shown in Supplemental Figure 7. Supply 12 V DC to the heater controller from the J4 terminal connected to the AC adapter. Decrease the voltage from 12 V DC to 3.3 V DC for the circuit power supply using a regulator because the CPU supply voltage is 3.3 V DC. 
	NOTE: USB 1 is a terminal for 5 V DC and serial signal of development PC. Although 5 V DC is not essential, it is used as the power source to program the CPU. This is also converted to 3.3 V DC by the regulator. J1 is a control signal terminal at the time of programming. This circuit is housed in the controller case shown in Supplemental Figure 8.</li>
	<li>Prepare a &#8220;heater control circuit&#8221; as shown in Supplementary Figure 7. Switch to 12 V DC with Q5 (P channel MOS FET) and supply it to the heater. Q5 is a switching element that controls 12 V DC with PWM to adjust the amount of power supplied to the heater. 
	NOTE: The circuit includes an LED to visually confirm that voltage is being supplied to the heater. This drive signal (HEATER_C) is a PWM signal from the CPU. When an overheat signal is detected by the protection circuit, the BREAKER signal switches to LOW, and the operation of the MOS-FET stops. This circuit is housed in the controller case shown in Supplemental Figure 8.</li>
	<li>Prepare a &#8220;connector circuit for heater unit&#8221; as shown in Supplementary Figure 7. Install a USB connector for connection with the heater section. 
	NOTE: This circuit is housed in the controller case shown in Supplemental Figure 8.</li>
	<li>Prepare a &#8220;connector circuit for temperature sensor&#8221; as shown in Supplemental Figure 7. Mount the connector (Euroblock receptacle 2P) to connect the temperature sensor. 
	NOTE: This circuit is housed in the controller case shown in Supplemental Figure 8.</li>
	<li>For a &#8220;A/D converter&#8221; as shown in Supplemental Figure 7, use ADS 1015 as an AD conversion device. 
	NOTE: AD conversion device converts the values of the temperature sensor and overheat detection sensor from voltage to digital values. This is a 12-bit multiplexer AD conversion device and is connected to the CPU with the I2C interface. This circuit is housed in the controller case shown in Supplemental Figure 8.</li>
	<li>Make a &#8220;protect circuit&#8221; as shown in Supplemental Figure 7 by connecting the overheat detection sensor (OHS) signal to the inverting input of the OP amp. Compare this signal with the voltage of the trimmer resistor connected to the noninverting input.
	<ol>
		<li>Ensure that when the voltage becomes lower than the voltage of the trimmer resistor, the output of the OP amplifier goes HIGH, the connected NPN transistor Q2 turns ON and the BREAKER signal goes LOW.</li>
		<li>Ensure that at the same time, Q4 turns ON and the connected overheat indicator LED D6 lights up. 
		NOTE: This circuit is housed in the controller case shown in Supplemental Figure 8.</li>
	</ol>
	</li>
	<li>For a &#8220;display section&#8221; as shown in Supplemental Figure 7, use 192 x 64 dots for OLED. Connect with the CPU via the I2C interface.
	<ol>
		<li>Reset the OLED by separating the GND of the OLED by the CPU signal IO0 using an NPN transistor Q1 connected to the GND of the OLED. 
		NOTE: This OLED displays various types of information. This circuit is housed in the controller case shown in Supplemental Figure 8.</li>
	</ol>
	</li>
	<li>For a &#8220;LED &#38; rotary encoder with push switch&#8221; in Supplemental Figure 7, mount a rotary encoder by solder that functions as a push switch and incorporates two LEDs.
	<ol>
		<li>Connect one LED to VCC for use as a power LED. The other is connected to the CPU for use as an indicator during heater operation.</li>
		<li>Use a push switch contact for heater START/STOP that is connected to CPU. Connect the A and B outputs of the rotary encoder to the IO input set in the CPU interrupt. 
		NOTE: This circuit is housed in the controller case shown in Supplemental Figure 8.</li>
	</ol>
	</li>
	<li>For the CPU in Supplemental Figure 7, use the CPU of WROOM - 02D.
	<ol>
		<li>Output from IO12, IO13 to the &#8220;display unit&#8221; because the interface of the display is I2C standard. Connect IO0 to &#34;display unit&#34; and reset the OLED.</li>
		<li>Connect IO15 to &#34;heater control unit&#34; and control the power supplied to the heater by PWM output.</li>
		<li>Connect IO2 to &#34;LED &#38; rotary encoder with push switch&#34; and light the START LED. Connect IO4 and IO14 to &#34;LED &#38; rotary encoder with push switch&#34; and receive the signals (REA and REB) from the rotary encoder to determine the set temperature. Connect IO5 to &#34;LED &#38; rotary encoder with push switch&#34; and start/stop the heater.</li>
	</ol>
	</li>
</ol>
3. Software design outline
<ol>
	<li>Use Arduino CORE for WROOM - 02D for the CPU as the controller for this system. 
	NOTE: As input devices, the start/stop switch, rotary encoder, temperature sensor (thermistor) are used. As output devices, an LED, character display (OLED), and heater are used. The communication device uses Wi-Fi.</li>
	<li>Outline of the operation
	<ol>
		<li>Detect the operation of the rotary encoder as shown in the LED &#38; rotary encoder with the push switch in Supplemental Figure 7, store it as the set temperature, and display it on the OLED. Set the input terminal of the CPU to which the phase terminals REA and REB are connected as an interrupt input terminal and process the rotation (forward and reverse) of the rotary encoder by interrupt. Set it to +1 for forward rotation and -1 for reverse rotation. Write the set temperature to the global variable and use it for the heater temperature control. At the same time, update the set temperature display of the OLED.</li>
		<li>Identify start and stop by CPU IO 5 by start/stop switch (SW-S) as shown in the CPU of Supplemental Figure 7. The state of the start/stop switch is a timer interrupt process every 50 ms. 
		NOTE: Since the switch is a momentary switch, it reverses the state of start/stop when it is pushed and released. This state is stored in the global variable.</li>
		<li>Use a thermistor for the temperature sensor. Read the measured values from the sample sensor (refer to &#8220;connector circuit for heater connection&#8221; in Supplemental Figure 7) into the CPU after A/D converter (refer to &#8220;A/D converter&#8221; in Supplemental Figure 7). Supply the current to the heater by turning on the IO15 port in the &#8220;CPU&#8221; of Supplemental Figure 7. 
		NOTE: There are two types of temperature sensors. One is used to measure the temperature of the sample and control the heater on the set temperature, and the other is attached to a heater and used for heat prevention. Connect the thermistor to 3.3 V via a resistor and record the change in resistance as a change in voltage. Remove a noise by the moving average method.</li>
		<li>Use a thermistor for temperature prevention temperature sensor. Overheat detection is performed using a thermistor (R2) (&#8220;connector circuit for heater connection&#8221; in Supplemental Figure 7), and when the set value is exceeded, the heater current is shut off (&#8220;protect circuit&#8221; in Supplemental Figure 7). 
		NOTE: This sensor is incorporated into a circuit and not through the CPU. This sensor is independent from the CPU and compared with the resistance value set by the resistor trimmer by a differential amplifier in an analog manner. When it detects that the temperature has exceeded the set value, it intervenes in the FET switch, which controls the current to the heater and forcibly stops the current supply. The purpose is to prevent the temperature of the heater from exceeding a certain level even in a situation where the CPU does not work properly.</li>
		<li>Turn on the LED in the &#8220;LED &#38; rotary encoder with push switch&#8221; of the Supplemental Figure 7 by the CPU (in the &#8220;CPU&#8221; of Supplemental Figure 7), when the equipment is in operation.</li>
		<li>Display the set temperature and measured value to OLED in the &#34;display section&#34; of the Supplemental Figure 7 by the CPU (in the &#34;CPU &#34;of Supplemental Figure 7).</li>
		<li>Drive the FET switch in the &#34;heater control circuit&#34; of Supplemental Figure 7 with PWM from CPU to control the heater.</li>
		<li>Control the heater by PID, based on measured temperatures acquired by the temperature sensor. Use Arduino&#39;s pid_v1.h library for PID processing. 
		NOTE: The CPU calibrates the time, communicates with the server, transmits data, and receives instructions from the server. When the sensor temperature exceeds the set temperature, the current to the heater is set to 0, and overshoot is suppressed.</li>
		<li>Use the built-in Wi-Fi connection function of the CPU and connect to the Internet. Transmit set the temperature, heater temperature, etc. to the designated server by Wi-Fi.</li>
	</ol>
	</li>
</ol>
4. System configuration
<ol>
	<li>Build the system according to Supplemental Figure 9.</li>
	<li>Equip a Wi-Fi with the controller.</li>
	<li>Use a thermistor as a sensor for temperature measurement. Connect the thermistor wire to the &#34;SENSOR&#34; terminal on the controller case. Receive the temperature signal measured by the thermistor.</li>
	<li>Connect a microscope stage incorporating the rubber heater and the &#34;HEATER&#34; of the controller case with a dedicated cable. Control the current to the rubber heater.</li>
	<li>Change the set temperature with the knob on the controller. 
	NOTE: Temperature log monitoring, temperature setting can be operated remotely from a PC or smartphone.</li>
	<li>Transfer the measured temperature, set temperature, and time information at measurement from the controller to the server via the internet. The data measurement cycle time is 5 s and cycle time for data transfer to the server is 1 min.</li>
	<li>Access the server from the controller side at regular intervals and transfer the measurement data stored in the CPU of the controller to the server for analysis and graphing.</li>
	<li>Refer to the supplementary material for how to operate the server.</li>
</ol>
5. Design of the sideways inverted microscope
<ol>
	<li>Fix two aluminum plates of 15 mm in thickness vertically with screws to create a basic mount.</li>
	<li>Attach a jig (one place) to the horizontal part of the base mount.</li>
	<li>Place the microscope stage part vertically, attach the jigs (two places) to the vertical part of the base stand and fix the bottom of the microscope to the base stand.</li>
	<li>Fix the microscope stage with screws.</li>
</ol>
6. Method of operation
NOTE: Here, the sample used is a mixture of Bold Modified Basal Freshwater Nutrient Solution liquid culture medium, sodium metasilicate, vitamins, and sterile water. 800 &#956;L of this sample is diluted in 10 mL of fresh water medium.
<ol>
	<li>Observation method
	<ol>
		<li>Inject 1,000 &#956;L of the prepared sample into a self-made glass chamber. 
		NOTE: The self-made glass chamber arranges two slide glasses in parallel and fixes them with an adhesive. A normal Petri dish has a large thickness and cells escape in the depth direction in the chamber, making it difficult to observe with a microscope. To prevent this, the chamber with a small depth direction is made, which makes it possible to prevent the cells from escaping in the depth direction in the chamber. A low temperature curable epoxy resin adhesive is used to bond around the glass to prevent the sample dropping from the chamber.</li>
		<li>Attach a separately prepared video camera to the microscope. Connect a video camera using the microscope&#39;s dedicated lens adapter and shoot the sample.</li>
		<li>Use a microscope with a 10x eyepiece and 200x objective.</li>
		<li>Attach the vertical microscope stage to a microscope in four locations with 4 mm screws. 
		NOTE: Refer to Supplemental Figure 1A and Supplemental Figure 2A for design drawings of aluminum plates. In this experiment, an inverted microscope was used. This was tilted by 90&#176;, and the fabricated microscope stage was affixed with screws. Refer to Figure 1.</li>
		<li>Secure the sample with two rubber bands using the four claws made lengthwise. Place a sample on a microscope stage perpendicular to the ground surface.</li>
		<li>Set the temperature to 40 &#176;C with the controller shown in Supplemental Figure 8. Turn the controller knob to set the temperature. Check the set temperature on the display. Press the knob to start temperature control, and the blue LED will light up. Press the knob again to turn off the LED and stop temperature control. 
		NOTE: The measured temperature is displayed in real-time, and the heater is controlled to reach the set temperature. When the temperature control starts, the blue LED lights up and remains so while the heater is in operation. When the heater overheats, the red LED lights, and the heater automatically stops.</li>
		<li>Refer to &#34;The Server Operation Manual&#34; in the supplementary information for server operation. 
		NOTE: A server for data storage is required. The server&#39;s database uses My-SQL.</li>
	</ol>
	</li>
</ol>
7. Measurement of surface temperature distribution of rubber heater
<ol>
	<li>Measure the distribution of the rubber heater surface temperature by thermography to check the temperature uniformity.</li>
	<li>Attach the microscope stage which incorporated a rubber heater with a stand.</li>
	<li>Change the setting temperature of the rubber heater surface to 35 &#176;C, 45 &#176;C, 55 &#176;C, and 65 &#176;C, and measure by the thermography from the front (refer to Supplemental Figure 10).</li>
</ol>
8. Temperature response test
<ol>
	<li>Start temperature control by setting the sample set temperature to 30 &#176;C. Wait until the measurement value reaches 30 &#176;C and stabilizes. Increase the preset temperature stepwise by 5 &#176;C from 30 &#176;C to 50 &#176;C and wait until the measured value stabilizes following the respective preset temperature.</li>
	<li>Decrease the preset temperature stepwise by 5 &#176;C from 50 &#176;C to 30 &#176;C and detect the tracking ability of the measured value.</li>
</ol>

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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diatom+Locomotion+-+An+Explanation.">Diatom Locomotion - An Explanation.</a> Protoplasma. 62, (1966).</li></ol>

The authors have no conflicts to disclose.

Trajectory analysis of moving diatom cells is a useful approach to evaluating diatom motility. However, while a normal inverted microscope observes samples horizontally, it is not suitable for observations of the influence of gravity or floating movement in the vertical direction. Developed and described here is a vertical microscope stage with temperature control and attached to an inverted microscope, which has been rotated by 90&#176;. This microscope stage with temperature control allows observation of temperature-de...

Optical microscopy is a technique employed to increase observable details via magnification of a sample with lenses and visible light. In optical microscopy, light is directed onto a sample, then transmitted, reflected, or fluorescent light is captured by magnifying lenses for observation. Various types of microscope are available that differ in design to accommodate different uses and observation methods. The different designs include an upright microscope, which is structured to illuminate a sample from below for observation from above, and an inverted microscope, which illuminates the sample from above for observation from below. Upright microscopes are the most common and widely used design. Inverted microscopes are often used to observe samples that cannot allow a lens close in distance from above, such as cultured cells adherent to the bottom of a container. Many research groups have reported observations in a wide range of fields using inverted microscopes1,2,3,4,5,6,7. Many additional devices have also been developed that take advantage of the features of inverted microscopes8,9,10,11,12,13.
Currently, in all conventional microscope designs, the microscope stage is horizontal and is therefore unsuitable for the observation of samples producing movement in the vertical plane, (due to gravity, buoyancy, motion, etc.). To make these observations possible, the microscope stage and light path must be rotated to vertical. The vertical stage is required to vertically mount glass slides or sample containers such as a Petri dishes to the stage. To address this, a sideways inverted microscope tilted by 90&#176; has already been devised. However, attaching samples with tape or other adhesives does not yield the necessary long-term immobility. Described here is a device that can achieve the necessary stability. This device permits observation over time of sample movement in the vertical plane. Mounting of a silicon rubber heater has also made it possible to observe the influence of temperature variation on sample behavior. Temperature data is transferred to an internet server by Wi-Fi, and temperature settings and log monitoring can be controlled remotely from a PC or smartphone. To our knowledge, the stage attached to a sideways tilted microscope tilted by 90&#176; has not yet been reported in previous studies.
The microscope stage is composed of three aluminum plates. The middle aluminum plate is mounted to the lower aluminum plate that attaches to the stage. The silicone rubber containing the temperature sensor is attached between the middle and upper aluminum plates. Rubber bands are used to affix the sample. Claws are attached in the left and right four points of the upper aluminum plate to secure the rubber bands. The control circuit of the temperature regulator receives a signal from the temperature sensor embedded in silicone rubber and modulates electric power by the pulse width modulation (PWM) method. The temperature can be gradually increased to 50 &#176;C in 1 &#176;C increments. This device is useful for applications in which vertical sample motions may be temperature-dependent.
This report provides examples of temperature effects on the floating phenomenon of diatoms. As examples of diatom observation studies, measurements of sedimentation velocity of cell clusters, motion analyses, ultrafine structure studies, etc. have been reported14,15,16,17,18,19,20,21,22,23. The specific gravity of diatoms floating in water with photosynthetic organisms is slightly higher than that of water, so they tend to sink; however, they will rise if even slight convection is occurring. To study this phenomenon, a glass slide is affixed vertically to a microscope stage, and the effects of increasing temperature on diatom vertical motion are observed.

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			<td>Inoval Co., Ltd.</td>
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			<td>Gifu, Japan</td>
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			<td>Inoval Co., Ltd.</td>
			<td>W 150mm&#215;L 200&#13212;&#215;T 5mm</td>
			<td>Gifu, Japan</td>
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			<td>Inoval Co., Ltd.</td>
			<td>D 100mm&#215;T 3mm (30&#934;)</td>
			<td>Gifu, Japan</td>
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			<td>Inoval Co., Ltd.</td>
			<td>D 100mm&#215;T 3mm (30&#934;)</td>
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			<td>Sigma-Aldrich Co. LLC</td>
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			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
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			<td>Tokyo, Japan</td>
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			<td>Akihabara Inc.</td>
			<td>M096P4W</td>
			<td>Tokyo, Japan</td>
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			<td>Tera Co., Ltd.</td>
			<td>W 42mm&#215;L 30&#13212;</td>
			<td>Ishikawa, Japan</td>
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			<td>Tera Co., Ltd.</td>
			<td>D 100mm&#215;T 0.5mm (20&#934;)</td>
			<td>Ishikawa, Japan</td>
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			<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>
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			<td>Three High Co., Ltd.</td>
			<td>D 100mm&#215;T 2.5mm (20&#934;)</td>
			<td>Kanagawa, Japan</td>
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			<td>Seeed Technology Co., Ltd.</td>
			<td>mh5.0</td>
			<td>Shenzhen, China</td>
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			<td>Akizuki Denshi Tsusho Co., Ltd.</td>
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fabrication of microscope stage for vertical observation with temperature control function

Samples are usually placed onto a horizontal microscope stage for microscopic observation. However, to observe the influence of gravity on a sample or study afloat behavior, it is necessary to make the microscope stage vertical. To accomplish this, a sideways inverted microscope tilted by 90&#176; has been devised. To observe samples with this microscope, sample containers such as Petri dishes or glass slides must be secured to the stage vertically. A device that can secure sample containers in place on a vertical microscope stage has been developed and is described here. Attachment of this device to the stage allows observation of sample dynamics in the vertical plane. The ability to regulate temperature using a silicone rubber heater also permits observation of temperature-dependent sample behaviors. Furthermore, the temperature data is transferred to an internet server. Temperature settings and log monitoring can be controlled remotely from a PC or smartphone.

Figure 2 shows the temperature distribution of the rubber heater. The surface temperature of the rubber heater was uniform at each temperature. Figure 3 shows the responsiveness of the measured temperature to set temperature changes. The orange line shows the set temperature and blue line shows the change of the sample temperature. The overshoot of the measured value to the setting change is small and the tracking is quick.
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Fabrication of Microscope Stage for Vertical Observation with Temperature Control Function

Herstellung von Mikroskopbühne für vertikale Beobachtung mit Temperaturregelungsfunktion

Presented here is a protocol using a temperature-controlled microscope stage that allows a sample container to be mounted on a vertical microscope.

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

fabrication-microscope-stage-for-vertical-observation-with

Research

JoVE Journal

Engineering

7.3K Aufrufe.  Tokyo University of Science. Dieses Gerät kann verwendet werden, um eine Probe auf einer vertikalen Mikroskopstufe zu fixieren, um die Beobachtung des Einflusses der Schwerkraft und die Arbeit an der Voninteresseer Probe zu ermöglichen. Die Befestigung dieses Geräts an der Bühne ermöglicht die Beobachtung der Probendynamik im vertikalen Rahmen und ermöglicht die Beobachtung temperaturabhängiger Probenverhalten. Um die Aluminiumplatten herzustellen, schneiden Sie mit einer Laserbearbeitungsmaschine ein 101-Millimet...

Serie: Fabrication of Microscope Stage for Vertical Observation with Temperature Control Function

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

Owing to its relativistic low-energy charge carriers, the interaction between graphene and various impurities leads to a wealth of new physics and degrees of freedom to control electronic devices. In particular, the behavior of graphene&#8217;s charge carriers in response to potentials from charged Coulomb impurities is predicted to differ significantly from that of most materials. Scanning tunneling microscopy (STM) and scanning tunneling spectroscopy (STS) can provide detailed information on both the spatial and energy dependence of graphene&#39;s electronic structure in the presence of a charged impurity.&#160;The design of a hybrid impurity-graphene device, fabricated using controlled deposition of impurities onto a back-gated graphene surface, has enabled several novel methods for controllably tuning graphene&#8217;s electronic properties.1-8 Electrostatic gating enables control of the charge carrier density in graphene and the ability to reversibly tune the charge2 and/or molecular5 states of an impurity. This paper outlines the process of fabricating a gate-tunable graphene device decorated with individual Coulomb impurities for combined STM/STS studies.2-5 These studies provide valuable insights into the underlying physics, as well as signposts for designing hybrid graphene devices.

This paper details the fabrication process of a gate-tunable graphene device, decorated with Coulomb impurities for scanning tunneling microscopy studies. Mapping the spatially dependent electronic structure of graphene in the presence of charged impurities unveils the unique behavior of its relativistic charge carriers in response to a local Coulomb potential.

Herstellung von Tor-abstimmbaren Graphene Geräte für Rastertunnelmikroskopie Studies mit Coulomb Verunreinigungen

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

Forschung

Ingenieurwesen

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

We demonstrate a method for the low temperature growth (350 &#176;C) of vertically-aligned carbon nanotubes (CNT) bundles on electrically conductive thin-films. Due to the low growth temperature, the process allows integration with modern low-&#954; dielectrics and some flexible substrates. The process is compatible with standard semiconductor fabrication, and a method for the fabrication of electrical 4-point probe test structures for vertical interconnect test structures is presented. Using scanning electron microscopy the morphology of the CNT bundles is investigated, which demonstrates vertical alignment of the CNT and can be used to tune the CNT growth time. With Raman spectroscopy the crystallinity of the CNT is investigated. It was found that the CNT have many defects, due to the low growth temperature. The electrical current-voltage measurements of the test vertical interconnects displays a linear response, indicating good ohmic contact was achieved between the CNT bundle and the top and bottom metal electrodes. The obtained resistivities of the CNT bundle are among the average values in the literature, while a record-low CNT growth temperature was used.

A method for the growth of low temperature vertically-aligned carbon nanotubes, and the subsequent fabrication of vertical interconnect electrical test structures using semiconductor fabrication is presented.

Herstellung von Niedertemperatur-Carbon Nanotube Vertikale Verbindungen Kompatibel mit Halbleitertechnik

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

Stretchable electronics are identified as a key technology for electronic applications in the next generation. One of the challenges in fabrication of stretchable electronic devices is the preparation of stretchable conductors with great mechanical stability. In this study, we developed a simple fabrication method to chemically solder the contact points between silver nanowire (AgNW) networks. AgNW nanomesh was first deposited on a glass slide via spray coating method. A reactive ink composed of silver nanoparticle (AgNPs) precursors was applied over the spray coated AgNW thin films. After heating for 40 min, AgNPs were preferentially generated over the nanowire junctions to solder the AgNW nanomesh, and reinforced the conducting network. The chemically modified AgNW thin film was then transferred to polyurethane (PU) substrates by casting method. The soldered AgNW thin films on PU exhibited no obvious change in electrical conductivity under stretching or rolling process with elongation strains up to 120%.

A simple synthesis method is used to chemically solder silver nanowire thin film to fabricate highly stretchable and conductive metal conductors.

Ein Herstellungsverfahren für hochverstreckbare Leiter mit Silber Nanodrähte

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

Liquid crystalline elastomers (LCEs) are smart materials capable of reversible shape-change in response to external stimuli, and have attracted researchers&#39; attention in many fields. Most of the studies focused on macroscopic LCE structures (films, fibers) and their miniaturization is still in its infancy. Recently developed lithography techniques, e.g., mask exposure and replica molding, only allow for creating 2D structures on LCE thin films. Direct laser writing (DLW) opens access to truly 3D fabrication in the microscopic scale. However, controlling the actuation topology and dynamics at the same length scale remains a challenge.
In this paper we report on a method to control the liquid crystal (LC) molecular alignment in the LCE microstructures of arbitrary three-dimensional shape. This was made possible by a combination of direct laser writing for both the LCE structures as well as for micrograting patterns inducing local LC alignment. Several types of grating patterns were used to introduce different LC alignments, which can be subsequently patterned into the LCE structures. This protocol allows one to obtain LCE microstructures with engineered alignments able to perform multiple opto-mechanical actuation, thus being capable of multiple functionalities. Applications can be foreseen in the fields of tunable photonics, micro-robotics, lab-on-chip technology and others.

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

Here, we fabricate 3D polymeric micro/nano structures in which both the shape and the molecular alignment can be engineered with nanometer scale accuracy by the use of direct laser writing. Light induced deformation of several types of liquid crystalline elastomer microstructures can be controlled in the microscopic scale.

Freiform-Lichtaktoren - Herstellung und Steuerung der Betätigung in mikroskopischer Skala

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

We demonstrate a method for the high temperature fabrication of porous, nanostructured yttria-stabilized-zirconia (YSZ, 8 mol% yttria - 92 mol% zirconia) scaffolds with tunable specific surface areas up to 80 m2&#183;g-1. An aqueous solution of a zirconium salt, yttrium salt, and glucose is mixed with propylene oxide (PO) to form a gel. The gel is dried under ambient conditions to form a xerogel. The xerogel is pressed into pellets and then sintered in an argon atmosphere. During sintering, a YSZ ceramic phase forms and the organic components decompose, leaving behind amorphous carbon. The carbon formed in situ serves as a hard template, preserving a high surface area YSZ nanomorphology at sintering temperature. The carbon is subsequently removed by oxidation in air at low temperature, resulting in a porous, nanostructured YSZ scaffold. The concentration of the carbon template and the final scaffold surface area can be systematically tuned by varying the glucose concentration in the gel synthesis. The carbon template concentration was quantified using thermogravimetric analysis (TGA), the surface area and pore size distribution was determined by physical adsorption measurements, and the morphology was characterized using scanning electron microscopy (SEM). Phase purity and crystallite size was determined using X-ray diffraction (XRD). This fabrication approach provides a novel, flexible platform for realizing unprecedented scaffold surface areas and nanomorphologies for ceramic-based electrochemical energy conversion applications, e.g. solid oxide fuel cell (SOFC) electrodes.

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

A protocol for fabricating porous, nanostructured yttria-stabilized-zirconia (YSZ) scaffolds at temperatures between 1,000 &#176;C and 1,400 &#176;C is presented.

Hochtemperatur Herstellung von Nanostrukturierte Yttrium-stabilisiertes Zirkonoxid (YSZ) Scaffolds durch<em&gt; In Situ</em&gt; Carbon-Templating Xerogele

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

We have demonstrated that through the sulfurization of transition metal films such as molybdenum (Mo) and tungsten (W), large-area and uniform transition metal dichalcogenides (TMDs) MoS2 and WS2 can be prepared on sapphire substrates. By controlling the metal film thicknesses, good layer number controllability, down to a single layer of TMDs, can be obtained using this growth technique. Based on the results obtained from the Mo film sulfurized under the sulfur deficient condition, there are two mechanisms of (a) planar MoS2 growth and (b) Mo oxide segregation observed during the sulfurization procedure. When the background sulfur is sufficient, planar TMD growth is the dominant growth mechanism, which will result in a uniform MoS2 film after the sulfurization procedure. If the background sulfur is deficient, Mo oxide segregation will be the dominant growth mechanism at the initial stage of the sulfurization procedure. In this case, the sample with Mo oxide clusters covered with few-layer MoS2 will be obtained. After sequential Mo deposition/sulfurization and W deposition/sulfurization procedures, vertical WS2/MoS2 hetero-structures are established using this growth technique. Raman peaks corresponding to WS2 and MoS2, respectively, and the identical layer number of the hetero-structure with the summation of individual 2D materials have confirmed the successful establishment of the vertical 2D crystal hetero-structure. After transferring the WS2/MoS2 film onto a SiO2/Si substrate with pre-patterned source/drain electrodes, a bottom-gate transistor is fabricated. Compared with the transistor with only MoS2 channels, the higher drain currents of the device with the WS2/MoS2 hetero-structure have exhibited that with the introduction of 2D crystal hetero-structures, superior device performance can be obtained. The results have revealed the potential of this growth technique for the practical application of 2D crystals.

Through the sulfurization of pre-deposited transition metals, large-area and vertical 2D crystal hetero-structures can be fabricated. The film transferring and device fabrication procedures are also demonstrated in this report.

Herstellung von großflächigen vertikale 2D Crystal Hetero-Strukturen durch Sulfurization Übergangsmetall Filme für Gerät Herstellung

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

Here, we demonstrate a method of coating ferromagnetic La0.67Sr0.33MnO3 (LSMO) nanoparticles on (001) SrTiO3 (STO) single-crystal substrates by radio frequency (RF) magnetron sputtering. LSMO nanoparticles were deposited with diameters from 10 to 20 nm and heights between 20 and 50 nm. At the same time, (Gd) Ba2Cu3O7&#8722;&#948; ((Gd) BCO) films were fabricated on both undecorated and LSMO nanoparticle decorated STO substrates using RF magnetron sputtering. This report also describes the properties of GdBa2Cu3O7&#8722;&#948;/ La0.67Sr0.33MnO3 quasi-bilayer films structures (e.g., crystalline phase, morphology, chemical composition); magnetization, magneto-transport, and superconducting transport properties were also evaluated.

Radio Frequency Magnetron Sputtering of GdBa2Cu3O7ÃƒÂ¢Ã‹â€ &#039;ÃƒÅ½ ´/ La0.67Sr0.33MnO3 Quasi-bilayer Films on SrTiO3 STO Single-crystal Substrates

Here, we present a protocol to grow LSMO nanoparticles and (Gd) BCO films on (001) SrTiO3 (STO) single-crystal substrates by radio frequency (RF)-sputtering.

Radiofrequenz das Magnetron-Sputtern GdBa2Cu3O7−δ/ La0,67Sr0,33MnO3 quasi-Bilayer Filme auf SrTiO3 (STO) Einkristall Substraten

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

We demonstrate extension of electron-beam lithography using conventional resists and pattern transfer processes to single-digit nanometer dimensions by employing an aberration-corrected scanning transmission electron microscope as the exposure tool. Here, we present results of single-digit nanometer patterning of two widely used electron-beam resists: poly (methyl methacrylate) and hydrogen silsesquioxane. The method achieves sub-5 nanometer features in poly (methyl methacrylate) and sub-10 nanometer resolution in hydrogen silsesquioxane. High-fidelity transfer of these patterns into target materials of choice can be performed using metal lift-off, plasma etch, and resist infiltration with organometallics.

We use an aberration-corrected scanning transmission electron microscope to define single-digit nanometer patterns in two widely-used electron-beam resists: poly (methyl methacrylate) and hydrogen silsesquioxane. Resist patterns can be replicated in target materials of choice with single-digit nanometer fidelity using liftoff, plasma etching, and resist infiltration by organometallics.

Einstelligen Nanometer Elektronenstrahl Lithographie mit einem Aberration korrigiert Scan Transmissions-Elektronenmikroskop

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

Soft pneumatic network actuators have become one of the most promising actuation devices in soft robotics which benefits from their large bending deformation and low input. However, their monotonous bending motion form in two-dimensional (2-D) space keeps them away from wide applications. This paper presents a detailed fabrication method of soft pneumatic network actuators with oblique chambers, to explore their motions in three-dimensional (3-D) space. The design of oblique chambers enables actuators with tunable coupled bending and twisting capabilities, which gives them the possibility to move dexterously in flexible manipulators, to become biologically inspired robots and medical devices. The fabrication process is based on the molding method, including the silicone elastomer preparation, chamber and base parts fabrication, actuator assembly, tubing connections, checks for leaks, and actuator repair. The fabrication method guarantees the rapid manufacturing of a series of actuators with only a few modifications in the molds. The test results show the high quality of the actuators and their prominent bending and twisting capabilities. Experiments of the gripper demonstrate the advantages of the development in adapting to objects with different diameters and providing sufficient friction.

Here we present a fabrication method of soft pneumatic network actuators with oblique chambers. The actuators are capable of generating coupled bending and twisting motions, which broadens their application in soft robotics.

Herstellung von weichen pneumatische Netzwerk Aktoren mit schrägen Kammern

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

Stimulated Raman scattering (SRS) microscopy uses near-infrared excitation light; therefore, it shares many multi-photon microscopic imaging properties. SRS imaging modality can be obtained using commercial laser-scanning microscopes by equipping with a non-descanned forward detector with proper bandpass filters and lock-in amplifier (LIA) detection scheme. A schematic layout of a typical SRS microscope includes the following: two pulsed laser beams, (i.e., the pump and probe directed in a scanning microscope), which must be overlapped in both space and time at the image plane, then focused by a microscope objective into the sample through two scanning mirrors (SMs), which raster the focal spot across an x-y plane. After interaction with the sample, transmitted output pulses are collected by an upper objective and measured by a forward detection system inserted in an inverted microscope. Pump pulses are removed by a stack of optical filters, whereas the probe pulses that are the result of the SRS process occurring in the focal volume of the specimen are measured by a photodiode (PD). The readout of the PD is demodulated by the LIA to extract the modulation depth. A two-dimensional (2D) image is obtained by synchronizing the forward detection unit with the microscope scanning unit. In this paper, the implementation of an SRS microscope is described and successfully demonstrated, as well as the reporting of label-free images of polystyrene beads with diameters of 3 &#181;m. It is worth noting that SRS microscopes are not commercially available, so in order to take advantage of these characteristics, the homemade construction is the only option. Since SRS microscopy is becoming popular in many fields, it is believed that this careful description of the SRS microscope implementation can be very useful for the scientific community.

In this manuscript, the implementation of a stimulated Raman scattering (SRS) microscope, obtained by the integration of an SRS experimental set-up with a laser scanning microscope, is described. The SRS microscope is based on two femtosecond (fs) laser sources, a Ti-Sapphire (Ti:Sa) and synchronized optical parametric oscillator (OPO).

Implementierung eines nichtlinearen Mikroskops auf Basis der stimulierten Raman-Streuung

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