Difference between revisions of "Part:BBa K515107:Experience"
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| + | <p><i>Figure 1: On the left, pictures of bacteria expressing <a href="https://parts.igem.org/Part:BBa_K515107">BBa_K515107</a>. 1 is the area photoconverted using the 405nm laser. 2 is an individual bacterium whose Dendra protein has undergone photoconversion. 3 is a negative control consisting of a non-photoconverted bacterium. On the right, graph representation of the photoconversion in the 3 marked areas. Intensity ROI 1 corresponds to area 1, Intensity ROI 2 corresponds to area 2, Intensity ROI 3 corresponds to area 3. Three different emmision spectra were observed, Ch2: emission in green spectrum. Ch3: emission in red spectrum. ChD: brightfield emission.</i></p> | ||
===User Reviews=== | ===User Reviews=== | ||
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Revision as of 07:28, 14 September 2011
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K515107
Characterisation
This part (BBa_K515107) has been characterised in a number of aspects to test its properties as a reporter. The tests describe this part in terms of thermostability, photostability and photoconversion.
Thermostability
Photostability
Photoconversion
Photoconversion using confocal microscope
This part has been used as a reporter for observation of bacterial uptake into the roots of the plants. Due to its photoconvertible properties, it allows to monitor the metabolic activity of the bacterial cell once uptaken into the root. Dendra was converted from 486nm excitation and 505nm emission wavelength, to 558nm excitation and 575nm emission wavelength using single photon stimulation. Conversion was achieved after exposure to 405nm wavelength using laser. Photoconversion was completed after about 15 rounds of bleaching at 50% laser intensity with the pinhole set to 3 airy units.
Figure 1: On the left, pictures of bacteria expressing BBa_K515107. 1 is the area photoconverted using the 405nm laser. 2 is an individual bacterium whose Dendra protein has undergone photoconversion. 3 is a negative control consisting of a non-photoconverted bacterium. On the right, graph representation of the photoconversion in the 3 marked areas. Intensity ROI 1 corresponds to area 1, Intensity ROI 2 corresponds to area 2, Intensity ROI 3 corresponds to area 3. Three different emmision spectra were observed, Ch2: emission in green spectrum. Ch3: emission in red spectrum. ChD: brightfield emission.
===User Reviews===