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Team:Freiburg/Content/Results/Light Boxes
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Revision as of 11:58, 17 October 2014
Our Light Boxes
One main principle of The AcCELLerator includes the illumination of our cells. This means we were in need of highly functional and easy to use light boxes.
Last year’s iGEM Team of Freiburg already examined the minimum requirements for a light box and designed an easy approach for building your own. Based on their ideas we constructed our improved boxes, which more sufficiently suited our own needs.
As we are experimenting with different light systems we build two red-light boxes, one with 660 nm and one with 740 nm, as well as a blue light box.
To ensure the specific wave length we were in need of, we merchandised diodes (Roithner Lasertechnik Germany, LED740-01AU/ LED660N-03) of the desired features and created an easy to replicate circuit to ensure straightforward operability. These circuit modules enable us to combine as many as required, to create the optimal conditions for high activation or inactivation efficiencies. Also it permits us to regulate the size of the light box more easily, so that it could fit at least three cell culture plates at a time for simultaneous experiments.
Fig. 1: Electronic circuit
Fig. 2: Functional diodes connected according our electronic circuit
Our boxes are made of simple fabrics from your local DIY-store, consisting of PVC plates and acrylic glass glued together and sealed. To ensure equally emitted light on any cell and reduction of the light intensity we also added the intensity decrease units, recommended by last year’s team of Freiburg (frosted glass, clear foil or white paper).
For our blue light system we were coincidently dependent on a functional light box and tried to improve the uniBOX of the iGEM Team Freiburg 2013 for our needs. We used the identical LED band as last year' team and build it into our own construct.
Fig. 3: Red Light Box
Fig. 4: Blue Light Box
Characterizations
Fig. 5: Wavelength measurement white light
Fig. 6: Wavelength measurement Light Box 660 nm
To ensure the same conditions we measured the light spectrum of our LED band similar to last year’s team and generated an equivalent result with only minor deviations. The blue light intensity in our box was measured 9 µmol/m²/s. Of course we were also interested in characterizing our red light boxes and tested the wave length of the build-in diodes by measuring the light spectrum.
Fig. 7: Wavelength measurement Light Box 740 nm
Especially for the red-light activation system we can guarantee the optimal wave length which is very important for us. The wave length of the far-red diodes is slightly shifted from the optimum but still sufficient for our experiments. We also measured the light intensities of the diodes and evaluated for the 660 nm diodes a value of 110 µmol/m²s, which is naturally toxic for our cell lines.
By adding the intensity decrease units we could reach a down regulation to 7 µmol/m²s or even down to 0.8 µmol/m²s, which suits best for the pattern experiments. The 740nm diodes are naturally less intense with 80 µmol/m²s and values down to 6 µmol/m²s and 0.5 µmol/m²s using different types of frosted glass.
