http://2012.igem.org/wiki/index.php?title=Team:Cambridge/Biologger/Labbook&feed=atom&action=historyTeam:Cambridge/Biologger/Labbook - Revision history2024-03-29T01:34:46ZRevision history for this page on the wikiMediaWiki 1.16.0http://2012.igem.org/wiki/index.php?title=Team:Cambridge/Biologger/Labbook&diff=297020&oldid=prevEmmyft at 03:10, 27 October 20122012-10-27T03:10:33Z<p></p>
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</table>Emmyfthttp://2012.igem.org/wiki/index.php?title=Team:Cambridge/Biologger/Labbook&diff=296992&oldid=prevEmmyft: Created page with "{{Template:Team:Cambridge/CAM_2012_TEMPLATE_HEADNEW}} <html> <script type="text/javascript"> $(document).keydown(function(e){ //e.which is set by jQuery for those browsers t..."2012-10-27T03:09:42Z<p>Created page with "{{Template:Team:Cambridge/CAM_2012_TEMPLATE_HEADNEW}} <html> <script type="text/javascript"> $(document).keydown(function(e){ //e.which is set by jQuery for those browsers t..."</p>
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= Biologger Labbook =<br />
<br />
==Week 3==<br />
<br />
===Wednesday (11/07/12)===<br />
<br />
'''Arduino circuitry'''<br />
<br />
----<br />
<br />
*Real-Time image captured by our freshly made primitive software, monitoring a light sensor on an arduino board. C++ was used to communicate with the Arduino and Python was used for data logging.<br />
<br />
===Friday (13/07/12)===<br />
<br />
'''Testing of the spectral properties of filters'''<br />
<br />
----<br />
<br />
*Blue , Green and Red filters were tested in different wavelengths (400 nm - 600 nm) in a spectrophotometer.<br />
<br />
*Excellent absorbance/emmitance results given by the blue and red filters near 580nm and 490nm respectively. 580nm is the wavelength where intensity of light is to be measured to identify the presence of mOrange. 490nm is the wavelength where maximum light intensity is expected with or without the presence of mOrange.<br />
<br />
[[File:MOrange_expected_graphs.jpg|250px|The expected output spectrum of our MOrange/luciferase fusion|thumb|left]]<br />
[[File:Filter_spectrum.png|250px|The spectral properties of our filters. The different colours represent the different colours of filter.|thumb|left]]<br />
<br />
==Week 4==<br />
<br />
===Friday (20/07/12)===<br />
<br />
'''Ratiometrica-Lux: Transformation of e.coli with lux genes from 2010'''<br />
<br />
----<br />
<br />
*Arabinose induction: Using the same 3mM arabinose prepared earlier in the week, the E. coli were spun down and the arabinose was added. After 5 hours, light is observed.<br />
*Testing the Arduino kit: the testing took place in the dark room, and the cell culture (in 1.5mL Eppendorf tubes) was held close to the photosensor and then taken away repeatedly. The data recorded was plotted with Matlab, as shown:<br />
[[File:CAM_Sensor_darkroom_glowingbacteria.jpg|thumb|700px|Light level against time]]<br />
<br />
*It should be noted that the background light levels were falling linearly, possibly due to negative feedback on the photosensor, but there is a consistent 5-6% increase of the original base reading upon placing the cells close the the photosensor.<br />
<br />
==Week 5==<br />
<br />
===Thursday (26/07/12)===<br />
<br />
'''Instrumentation: Hardware implementation'''<br />
<br />
----<br />
<br />
[[File:deviceprototype.jpg|thumb|500px|Our device, reaching perfection!]]<br />
<br />
<br />
<br />
Our device is close to being implemented. The filtered photoresistors are placed now in the right position, the motory circuit is also functioning, the main chassis of the device is constructed and painted. However, the mirrors which will be used to surround the cuvettes are still missing and so is the mechanical coupler designed by us and being constructed at the Instrument shop of the Engineering Department. The device is also powered by a 9V battery, without the need to be connected to the computer.<br />
<br />
==Week 14==<br />
<br />
===Monday (24/09/12)===<br />
'''Instrumentation'''<br />
<br />
The sensitivity of the sensor concerning light intensities was tested using a dilution series of luciferase-producing bacteria. 20ml Cultures were grown overnight from single colonies. The cultures were induced with 40ul of 1.5M arabinose (for a final concentration of 3mM). Cultures were left for 2 1/2 hours for full induction. Subsequently, a culture was pelleted and resuspended in 4ml LB. Doubling dilutions, of volume 2ml, were made from this concentrate, down to 1/8th concentration.<br />
1ml of each 2ml dilution was analysed in each cuvette, which was placed in the cuvette holder we made ourselves. <br />
<br />
[[File:Dilutiontest.jpg|thumb|400px|Raw sensor values(V) vs Concentration of bioluminescent E.coli- Concentrations on the x-axis are relative therefore an OD 600 value was also taken|center]]<br />
<br />
[[File:Norm_light.png|thumb|400px|Normalised sensor data using a dilution series of bioluminescent E.coli- Concentrations on the x-axis are relative therefore an OD 600 value was also taken|center]]<br />
<br />
[[File:Norm_blue.jpg|thumb|400px|Normalised percentage of blue frequencies using the same dilution series of E.coli|center]]<br />
<br />
[[File:Dilution_series.jpg|thumb|400px|Dilution series of E.coli from most concentrated to least|center]]<br />
<br />
===Tuesday (25/09/12)===<br />
<br />
'''Instrumentation'''<br />
<br />
The sensitivity of the sensor concerning different frequencies (colours) of light was tested as well. As can be seen below, measurements taken from orange and blue light yield values respectively above and below those from white light (our reference point). The data was taken using a constant intensity of light for each case (V.High and V.Low brightness). This was done with the aid of an Android phone and a specialised software application, called Color Flashlight, downloaded from the official Market. <br />
[[File:Sensor_colour.jpg|400px|Sensor data for different colours at different intensities|thumb|center]]<br />
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