Team:Grenoble-EMSE-LSU/Project/Monitoring/Cell2Machine

From 2013.igem.org

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<h1>Cell to Machine Communication</h1>
<h1>Cell to Machine Communication</h1>
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<p>To create a means of communication from cell to machine, we chose the red fluorescence protein KillerRed as a reporter protein. The first condition that our device needs to fulfill is to record light intensity. Then it needs to generate fluorescence thanks to a light source and a couple of excitation and emission filters. Firstly we will explain the choice of <a href="#Component">the different components</a>, then the several experiences we did to find the most accurate parameters for each part of the device – <a href="#Photodiode">the photodiode</a>, <a href="#Arduino">Arduino</a> and <a href="#Optic">the optic</a>.</p>
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<p>To create a means of communication from cell to machine, we chose the red fluorescence protein KillerRed as a reporter protein. The first condition that our device needs to fulfill is to record light intensity. Then it needs to generate fluorescence thanks to a light source and a couple of excitation and emission filters. First we will explain how we chose<a href="#Component">the different components</a>, then the experiments we did to find the most accurate parameters for each part of the device – <a href="#Photodiode">the photodiode</a>, <a href="#Arduino">Arduino</a> and <a href="#Optic">the optical components</a>.</p>
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<h2 id="Component">The components</h2>
<h2 id="Component">The components</h2>
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<p>To record light intensity, we were inspired by the E. glometer of the               Cambridge team of iGEM 2010.<br>
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<p>To record light intensity, we were inspired by the E. glometer of the Cambridge team of iGEM 2010.<br>
                                         <p align="center"><img src="https://static.igem.org/mediawiki/2013/a/a7/Eglometer.png" alt="The Eglometer" width="500px" /></p>
                                         <p align="center"><img src="https://static.igem.org/mediawiki/2013/a/a7/Eglometer.png" alt="The Eglometer" width="500px" /></p>
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We use quite the same photodiode (TSL230RD) – same as the TSL230RP-LF but in surface mounted device (SMD) – and an Arduino Uno. Arduino is a single-board microcontroller created to make electronics more accessible. The main asset of the photodiode is that the output can be either a pulse train or a square wave (50% duty cycle) with frequency directly proportional to light intensity. Since we are using a microcontroller, it is easy to calculate a frequency with the digital input of the microchip thanks to high or low level and we will have a better resolution because low frequencies are easier to measure than low voltages. For the optic part we use a domestic LED lamp and a cube filter from a microscope with excitation and emission filters and an adjustable lens. A domestic LED lamp was chosen to allow us not to buy several high-power LEDS and built a card with a heat sink. This lamp illuminate with 520 lumens in a cone of 40° under 12V and 6W. The low voltage was chosen as a safety condition and the small angle to avoid losing to much light. The excitation filter is a green interferential filter to excite the red fluorescent protein and the red emission filter is only a colored filter to collect all the red light in order to have a more efficient measure. In the cube there is also a dichroic mirror that reflects all the green light and transmits all the red light. This mirror enables us to separate completely the photodiode from the light source.
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We use a similar photodiode (TSL230RD) – the same as the TSL230RP-LF but as a surface mounted device (SMD) – and an Arduino Uno. Arduino is a single-board microcontroller created to make electronics more accessible. The main asset of the photodiode is that the output can be either a pulse train or a square wave (50% duty cycle) with its frequency directly proportional to light intensity. Since we are using a microcontroller, it is easy to calculate the frequency with the digital input of the microchip thanks to high or low level detection and we will have a better resolution because low frequencies are easier to measure than low voltages at low light levels. For the optical part we use a LED lamp and a cube filter from a fluorescence microscope with excitation and emission filters and an adjustable lens. The LED lamp was chosen so that e didn't have to buy high-power LEDS and build a card with heat sinks. This lamp illuminates with 520 lumens in a 40° cone under 12V and 6W. The low voltage was chosen as a safety measure and the small angle to avoid losing too much light. The excitation filter is a green interferential filter to excite the red fluorescent protein and the red emission filter is only a colored filter to collect all the red light in order to have a more precise measure. In the cube there is also a dichroic mirror that reflects all the green light and transmits all the red light. This mirror enables us to separate the photodiode from the light source completely.
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<h2 id="Photodiode">The photodiode</h2>
<h2 id="Photodiode">The photodiode</h2>
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<p>Before measuring with the photodiode, we need to know if the photodiode works as indicated in the datasheet. The photodiode was plug on a 5V stabilized power supply.</br></p>
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<p>Before measuring with the photodiode, we need to know if the photodiode works as indicated in the datasheet. The photodiode was plugged in on a 5V stabilized power supply.</br></p>
                                         <p align="center", style="margin:20px"><img src="https://static.igem.org/mediawiki/2013/5/5f/IGEMerworkphotodiode.png" alt="memberworkingonphotodiode" width="500px"></p>
                                         <p align="center", style="margin:20px"><img src="https://static.igem.org/mediawiki/2013/5/5f/IGEMerworkphotodiode.png" alt="memberworkingonphotodiode" width="500px"></p>

Latest revision as of 20:13, 16 September 2013

Grenoble-EMSE-LSU, iGEM


Grenoble-EMSE-LSU, iGEM

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