Team:Grenoble-EMSE-LSU/Project/Instrumentation/Fluo

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                                         <p>For the proof of concept of the optical part we use a LED lamp - MR16 (GU5.3)- and a cube filter from a fluorescence microscope with excitation and emission filters and an adjustable lens. The LED lamp was chosen so that we didn't have to buy <strong>high-power LEDS</strong> and build a <strong>card with heat sinks</strong>. This lamp illuminates with <strong>520 lumens in a 40° cone under 12V and 6W</strong>. The low voltage was chosen as <strong>a safety measure</strong> and the small angle to <strong>avoid losing too much light</strong>. The excitation filter is a <strong>green interferential filter</strong> to excite the red fluorescent protein and the <strong>red emission filter</strong> 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 <strong>dichroic mirror</strong> that reflects all the green light and transmits all the red light. This mirror enables us to <strong>separate the photodiode from the light source completely</strong>.</br></br></p>
                                         <p>For the proof of concept of the optical part we use a LED lamp - MR16 (GU5.3)- and a cube filter from a fluorescence microscope with excitation and emission filters and an adjustable lens. The LED lamp was chosen so that we didn't have to buy <strong>high-power LEDS</strong> and build a <strong>card with heat sinks</strong>. This lamp illuminates with <strong>520 lumens in a 40° cone under 12V and 6W</strong>. The low voltage was chosen as <strong>a safety measure</strong> and the small angle to <strong>avoid losing too much light</strong>. The excitation filter is a <strong>green interferential filter</strong> to excite the red fluorescent protein and the <strong>red emission filter</strong> 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 <strong>dichroic mirror</strong> that reflects all the green light and transmits all the red light. This mirror enables us to <strong>separate the photodiode from the light source completely</strong>.</br></br></p>
                                         <p align="center"><img src="https://static.igem.org/mediawiki/2013/c/ca/Optique.png" alt="Fluorometer_igem2013_Grenoble-EMSE-LSU" width="600px" /></p>
                                         <p align="center"><img src="https://static.igem.org/mediawiki/2013/c/ca/Optique.png" alt="Fluorometer_igem2013_Grenoble-EMSE-LSU" width="600px" /></p>
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                                         <p id="legend">Figure 5.<br>Talk'E.coli: C2M part
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                                         <p id="legend">Figure 5.<br>TalkE'coli: C2M part
On the left: the real device, on the right: functional scheme</br>
On the left: the real device, on the right: functional scheme</br>
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The light from the LED lamp goes through the green excitation filter and illuminateS the sample thanks to a dichroic mirror. Then the red fluorescent protein is now excited and re-emits red light that goes through a lens that concentrateS it on the photodiode.</br></br>  
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The light from the LED lamp goes through the green excitation filter and illuminates the sample thanks to a dichroic mirror. Then the red fluorescent protein is now excited and re-emits red light that goes through a lens that concentrates it on the photodiode.</br></br>  
</p>
</p>
<p>With the setup shown above, we put different culture in a 50mL rounded tube and to protect the photodiode from the outside lamp we place all the component in a large box. These are the results we obtained:</br></br></p>
<p>With the setup shown above, we put different culture in a 50mL rounded tube and to protect the photodiode from the outside lamp we place all the component in a large box. These are the results we obtained:</br></br></p>
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<p align="center"></br></br><img src="https://static.igem.org/mediawiki/2013/0/0a/Charac_fluo_measure.png" alt="Charac_fluo_measure" width="600px" /></br></br></p>
<p align="center"></br></br><img src="https://static.igem.org/mediawiki/2013/0/0a/Charac_fluo_measure.png" alt="Charac_fluo_measure" width="600px" /></br></br></p>
<p id="legend">Figure 6.<br>Characterization of the fluorescence measurements</p>
<p id="legend">Figure 6.<br>Characterization of the fluorescence measurements</p>
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<p>The fluorescence readings of Talk'E.coli and of the Tristar microplate reader are <strong>linearly related</strong>. Furthermore, the precision of both measurements are comparable. Our device is therefore <strong>able to detect KillerRed fluorescence with enough precision</strong> to allow proper cell growth control.
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<p>The fluorescence readings of TalkE'coli and of the Tristar microplate reader are <strong>linearly related</strong>. Furthermore, the precision of both measurements are comparable. Our device is therefore <strong>able to detect KillerRed fluorescence with enough precision</strong> to allow proper cell growth control.
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                                         </p>
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</br>The MOS transistor is controlled by Arduino and is used like a switch. It allows us to control the average light intensity of the LED lamp.</br></br>                                       
</br>The MOS transistor is controlled by Arduino and is used like a switch. It allows us to control the average light intensity of the LED lamp.</br></br>                                       
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<p>The first part of this circuit – all components above the MOS transistor BS170 - stabilizes the current of the LED bulb and the second part – consisting in the MOS transistor and Arduino microcontroller- allows us to control the average light intensity.</br>
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<p>The first part of this circuit – all components above the MOS transistor BS170 - <strong>stabilizes the current of the LED lamp</strong> and the second part – consisting in the MOS transistor and Arduino microcontroller- allows us to <strong>control the average light intensity</strong>.</br>
The nominal power of the LED is 6W when 12V is applied. That means that the current through the LED lamp is 0.5A.</br>  
The nominal power of the LED is 6W when 12V is applied. That means that the current through the LED lamp is 0.5A.</br>  
To ensure that the power supply is stable enough, we stabilized it thanks to a bipolar transistor, three diodes and two resistors.</br></br>  
To ensure that the power supply is stable enough, we stabilized it thanks to a bipolar transistor, three diodes and two resistors.</br></br>  
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<img src="https://static.igem.org/mediawiki/2013/a/a8/Servo_pos2.png" alt="Position2_servo" width="450px" /></p>
<img src="https://static.igem.org/mediawiki/2013/a/a8/Servo_pos2.png" alt="Position2_servo" width="450px" /></p>
<p id="legend">Figure 9.<br>On the left, the first position of the servomotor and on the right, the second position of the servomotor.</br>
<p id="legend">Figure 9.<br>On the left, the first position of the servomotor and on the right, the second position of the servomotor.</br>
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Known dimensions :</br></p>
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</p>
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<p align="left"><strong>L</strong>: distance between the center of the servomotor S and the center of the hole in the box A (6.5cm)</br>
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<p align="left">Known dimensions :</br>
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<strong>L</strong>: distance between the center of the servomotor S and the center of the hole in the box A (6.5cm)</br>
<strong>h</strong>: height from A to S (2cm)</br>
<strong>h</strong>: height from A to S (2cm)</br>
<strong>R</strong>: radius of the filter and also the hole in the box (1cm)</br>
<strong>R</strong>: radius of the filter and also the hole in the box (1cm)</br>

Latest revision as of 03:10, 5 October 2013

Grenoble-EMSE-LSU, iGEM


Grenoble-EMSE-LSU, iGEM

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