Team:Valencia/Hardware

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Latest revision as of 03:23, 22 October 2009

LEC activation

Our team prides itself in finding ways to link electronics to biology in a direct and innovative fashion. As a consequence we spent considerable time developing circuits and equipment allowing for quick and robust control of cellular physiology.

Electrostimulation for yeast

Our team has developed an electronic circuit capable of applying a voltage between 0 V and 24 V with a precision of 0.1 volts and during time intervals of up to 20 ms. to yeast cultures. This circuit has been used in a first stage to characterize the part [http://partsregistry.org/wiki/index.php?title=Part:BBa_K222000 BBa_K222000] or what we call LEC. In the characterization, this system supplies the initial estimulation conditions for the cells for the different experiments very precisely, making the results obtained with the luminometer of great confidence.

The experimental device is composed of the following parts:

The first one is made of a voltage source and two electrodes to stimulate the cultures.

The second one is an optocoupler performing the function of an electronic switch. It is composed of a led diode and a phototransistor. The phototransistor will allow to voltage signal while the diode is illuminated.

The last part is the responsible of the fine control of the time period in which the voltage is applied. To do that we stimulate the diode with a PC sound card (it has a precise voltage signal that is transformed by the loudspeakers to reproduce sound) controlled by the function “sound” of the MatLab software.

The output signal of the sound card was modified by a rectifier-filter and then passed through two operational amplifiers (one as an amplifier and the other as a tension follower) in order to produce a very precise signal able to activate the optocoupler. The amplifiers are powered by 5 volts and ground, so we will have 5 volts at the output of the operational when the sound card is active, and 0 volts when idle.

This design allows the stimulus of one channel, one LEC. In order to make a working screen, we will need an array of electrodes connected to a multiplexer. For that, see the next section...

Retrieved from "http://2009.igem.org/Team:Valencia/Hardware"

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-<div align="justify" style="position:relative; top:-5px; left:70px; width:600px">
+<br>
-=='''Experimental Set Up''' ==
+<div align="justify" style="position:relative; margin-top:-265px; margin-left:190px; width:700px; font-size:10pt; font-family: Verdana">
-<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">[under construction. Feel free to read an article while its being developed]
+=='''LEC activation'''==
+<br>
-<span style="color:black; align:justify; font-size:10pt; font-family: Verdana">
+Our team prides itself in '''finding ways to link electronics to biology in a direct and innovative fashion.''' As a consequence we spent considerable time developing circuits and equipment allowing for quick and robust control of cellular physiology.
-<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">Our team prides itself in finding ways to link electronics to biology in a direct and innovative fashion. As a consequence we spent considerable time developing circuits and equipment allowing for quick and robust control of cellular physiology.
 <br><br>
-*<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">'''Direct electrical stimulation of cells'''
+<!-- cite -->
+<br>
+==='''Electrostimulation for yeast'''===
-<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">The first part of hardware design is an electronic amplifier capable of delivering range of voltages between 0 and 13 volts. It is based on an inverting amplifier (fig. 1).
+Our team has developed '''an electronic circuit''' capable of applying a '''voltage between 0 V and 24 V with a precision of 0.1 volts and during time intervals of up to 20 ms.''' to yeast cultures. This circuit has been used in a first stage to characterize the part [http://partsregistry.org/wiki/index.php?title=Part:BBa_K222000 BBa_K222000] or what we call LEC. In the characterization, this system supplies the initial estimulation conditions for the cells for [https://2009.igem.org/Team:Valencia/WetLab/YeastTeam/Results the different experiments] very precisely, making the results obtained with the luminometer of great confidence.<br>
-The input voltage comes from a sound card – a commonly accessible and low cost digital to analog converter. It allows us to control the waveform in an arbitrary fashion using a simple Matlab script.
-[[Image:V_SoundCircuit.jpg|200px|center]]
+The experimental device is composed of the following parts:<br>
-<!-- [photo of the device connected to a sound card] -->
+*The first one is made of '''a voltage source and two electrodes''' to stimulate the cultures.
-<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">The output is connected to platinum electrodes inserted into a buffer over either muscular, neuronal or yeast cells.  We stimulate cells with delta-function pulses every second (or a few of them) to get continuous calcium influx. While muscular and neuronal cells respond to this kind of stimulus naturally, yeast will require some work. We hope that electrical stimulation of yeast could be obtained thanks to a heat-shock response (read more in modeling section) or reported calcium response to electroporation.
+*The second one is '''an optocoupler performing the function of an electronic switch'''. It is composed of a led diode and a phototransistor. The phototransistor will allow to voltage signal while the diode is illuminated.
-<!-- cite -->
+*The last part is '''the responsible of the fine control of the time period in which the voltage is applied'''. To do that we stimulate the diode with a PC sound card (it has a precise voltage signal that is transformed by the loudspeakers to reproduce sound) '''controlled by the function “sound” of the MatLab software'''.<br>
-<br>
+The output signal of the sound card was modified by a rectifier-filter and then passed through two operational amplifiers (one as an amplifier and the other as a tension follower) in order to produce '''a very precise signal able to activate the optocoupler'''. The amplifiers are powered by 5 volts and ground, so we will have 5 volts at the output of the operational when the sound card is active, and 0 volts when idle.
-*<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">'''Electrostimulation for yeast'''
-<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">Our team has developed '''an electronic circuit''' capable of sending electrical impulses to yeast in a large range of voltages [0-100] V and high accuracy at the time of application of these, as they can reach a few milliseconds. As our initial design for neurons and muscles (which was more restricted in terms of voltages and powers), this circuit only serves '''to control a pixel independently''', but it can easily be '''controlled from any computer'''.<br> <br>
+[[Image: V_ElectroYeast.png|700px]]
-In this circuit there are two well differentiated parts: ''the application part'', which highlights an adjustable voltage source and the electrodes through which we will stimulate our cells, and ''the control''.<br> <br>
-Both are ''linked by an optocoupler'', which serves as the electronic switch. It consists of one LED (on the control) and a phototransistor (part of application), which lets flow current while the collector-emitter diode is lit. To stimulate the diode controlling the time of application accurately, '''we use the sound card''' (which has a high frequency) led by the "sound" Matlab function. <br> <br>
-But the nature of sound is sinusoidal and therefore also the tension leaves the sound card, and we need to incorporate into our circuit ''a rectifier-filter control'', to ensure that our signal is as continuous as possible. Moreover, the tension that provides the sound card is insufficient to activate the optocoupler. This is the reason why we use ''two operational amplifiers'', one as amplifier and another simply as a voltage follower. The amplifiers are powered by 5 volts and ground, so we will have 5 volts at the output of the operational when the sound card is active, and 0 volts when idle.
-[[Image: V_ElectroYeast.png|625px]]
 <br><br>
-<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">This design only allows for stimulating one channel at once. While it is useful for proof-of-principle experiments, if we want to make a working screen, we will need an array of electrodes connected to a multiplexer. We are currently developing a device capable of stimulating tens of pixels at once.<br><br>
+'''This design allows the stimulus of one channel, one LEC. In order to make a working screen, we will need an array of electrodes connected to a multiplexer'''. For that, see [https://2009.igem.org/Team:Valencia/Hardware/iLCD the next section...]<br><br>
-*<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">'''Yeast screen'''
-<span style="color:black; align:justify; font-size:11pt; font-family: Verdana">Having established that yeast responds with light to electrical impulses, we consider to control an array of 96 pixels totally independent, so we can create '''moving figures''' and it would be the first screen that works with living cells.<br> <br>
-To do this we had two problems, to control each pixel independently and build a stand with 96 pairs of electrodes (one for each pixel) to stimulate the yeast.<br><br>
-) To '''control''' 96 pixels.
-Our screen is a matrix of 8 columns by 12 rows. Thus, we can decide which row and column we want to activate at all times through a card with 20 digital outputs, one per row and column. For example, to turn the cell in position 3,4 of the screen, activate the outputs corresponding to row 3 and column 4. The card is controlled by a program executed in LabVIEW.<br><br>
-[[Image: V_ScreenCircuit.jpg|400px|center]]
-<br>
-) '''Costruction''' of the support.
-<br><br><br><br>