These edit formats contain of glass masked with black tape containing specific gaps. To gain a spatial resolution different edit formats were used.

After an incubation of 1 h in dark the plate was illuminated with red light at an intensity of 0.02 - 0.5 ♞ for 15 - 60 min.Again culture medium was exchanged after 24 h with culture medium containing 15 µM of PCB under safe green light.A change of culture medium was performed 4 h post transfection.After 24 h the cells were transfected with 12 µg of tetR-pif and vp16-phyB and 8 µg of mcherry reporter.3.5*10^6 CHO-K1 cells were spread in a 10 cm culture dish.This protocoll was adapted from Müller et al., 2013. All following experiments were done with chinese hamster ovary cells (CHO-K1 cells). Therefore after an illumination with 660 nm the gene activation domain VP16 will be brought to TetR resulting in an expression of mCherry. TetR is able to bind to a TetO DNA target side that was cloned in front of a CMV minimal promotor driving an mCherry reporter gene. To get more information about the general functionality of the PhyB - PIF red light system klick here. 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) (Müller et al., 2013). The light intensity in general can be measured using a radiometer. The appropriate place to add these papers was described here. For very small light intensities you can use normal paper instead of baking paper. Using the uniBOX you can do this by adding layers of baking paper between the LEDs and your samples. Nearly every common green LED can be used to produce safe green light.įor blue light experiments red LEDs can be used to produce safe light conditions.įor some light experiments it may be of great importance to regulate the light intensity.

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

2 LED light chains (one for the light box and one for the 'dark' room to get your save light conditions).

An incubator (also impervious to light in the optimal case).

A room that can be made impervious to light (even a store room would serve perfectly well).

A common box to keep the samples in dark.

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

Unibox exchange