Illumination solutions for Uncaging Experiments

Caged compounds act as light-sensitive probes that functionally encapsulate biomolecules rendering them inactive. By applying targeted illumination, it is possible to “uncage”, that is to release and activate the biomolecules within at a precise time point, and then study the effects that follow. This can be a very powerful means to study the localized effects of dynamic biological processes such as signalling pathways. Illumination within the blue to UV spectrum is typically used to uncage the biomolecules and initiate the experiment, coupled with fluorescence microscopy to permit the observations that follow. The illumination intensity is typically at a higher power density than is used for photoactivation, photoswitching or photoconversion experiments.

Uncaging  Illumination Solutions

The precise, timed control of illumination power and wavelength of illumination to region(s) of interest is a vital part of uncaging biomolecules. Mosaic and MicroPoint are two products that are suitable for photostimulation experiments, each having some benefits that suit particular usage scenarios.

Selected Publications for Uncaging 

• Ryanodine receptor-mediated Ca2+ release and atlastin-2 GTPase activity contribute to IP3-induced dendritic Ca2+ signals in primary hippocampal neurons. Omar A. Ramírez, Alex Córdova, Mauricio Cerda, Pedro Lobos, Steffen Härtel, Andrés Couve, Cecilia Hidalgo, Cell Calcium, Volume 96, 2021, 102399, ISSN 0143-4160, https://doi.org/10.1016/j.ceca.2021.102399 ​
• ZipSeq: barcoding for real-time mapping of single cell transcriptomes. Hu, K.H., Eichorst, J.P., McGinnis, C.S. et al. Nat Methods 17, 833–843 (2020). https://doi.org/10.1038/s41592-020-0880-2
• Optogenetic control of RhoA to probe subcellular mechanochemical circuitry. Cavanaugh, K. E., Oakes, P. W., & Gardel, M. L. (2020). Current Protocols in Cell Biology, 86, e102. doi/10.1002/cpcb.102 ​
• Measurement of Microtubule Half-Life and Poleward Flux in the Mitotic Spindle by Photoactivation of Fluorescent Tubulin. Girão H., Maiato H. (2020), In: Maiato H. (eds) Cytoskeleton Dynamics. Methods in Molecular Biology, vol 2101. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0219-5_15 ​