Troubleshooting

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.