Many times science can be 1% inspiration, 99% iteration. Part of getting good results is reviewing your less-than-perfect outcomes and figuring out what to try next time. Learn about some of the problems you may encounter, see examples of what it looks like when things go wrong, and get hints for preventing those same problems in the future.
Background fluorescence
Learn ways to reduce background (or noise)—the nonspecific fluorescent signal coming from ambient light, the microscope, or the experiment components.
Photobleaching
Fluorescence that starts out bright but dims over time may be because the fluorophore is being degraded by light (photobleaching). Learn how to minimize it.
Bleed-through
When two or more fluorophores are used, light from one fluorophore can “contaminate” the channel of the other. Learn ways to minimize this bleed-through.
Phototoxicity
Learn the sources of phototoxicity, how to recognize stressed cells, and ways to reduce this kind of damage in your live-cell imaging experiments.
Uneven illumination
See an example of how a misaligned microscope light path can lead to variability in brightness in your stained cell sample.
For Research Use Only. Not for use in diagnostic procedures.