Team:Freiburg/Project/unibox

uniBOX - A lightbox build from lego!

What you need

Using the light box and performing light experiments is easy and can be done in every lab. All you need for your experiments is the following:

• The uniBOX (see construction)
• A common box to keep the samples in dark
• A room that can be made impervious to light (even a storeroom would serve perfectly well)
• An incubator (also impervious to light in the optimal case)
• 2 LED light chains (one for the light box and one for the "dark" roam to get your save light conditions)
• A radiometer to measure light intensities

Principles of light experiments

In principle every light experiment works similar. All you have to do is to proceed your samples under safe light in the dark room. Otherwise it is indispensable to keep your samples in the uniBOX or dark box to protect them from sunlight and artificial light. Even inside of the incubator this protection is important. Safe light are light conditions that do not affect your light experiments. That means if you are doing a red light experiment it is important that the safelight do not include wavelengths form 600 to 700 nm. Therefore the safe light conditions for red light can be green light or blue light. As green light contains less energy it is probably the best choice. Nearly every common green LED can be used to produce safe green light.

Set up the light box

For some light experiments it may be of great importance to regulate the light intensity. Using the uniBOX you can do this by adding layers of baking paper between the LEDs and your samples. For very small light intensities you can use normal paper instead of baking paper. The appropriate place to add these papers was described here . The light intensity in general can be measured using a radiometer.

To test our light Box (uniBOX) we performed a spatio-temporal gene expression experiment by using a red light inducible Tet-Off system (TetR fused to PIF and VP16 fused to PhyB) generously provided by Konrad Müller. To get more information about the general functionality of the PhyB - PIF red light system klich here. TetR is able to bind to a TetO DNA target side that was cloned in front of a CMV minimal promotor driving a mCherry reporter gene. Therefore after an illumination with 660nm the gene activation domain VP16 will be brought to TetR resulting in an expression of mCherry.

Method - Spatio-temporal gene expression