21 – LSM 880 Training Guide
Understanding Image Quality
Laser Intensity
The overall power of the laser will directly affect image quality but most
importantly it will have the greatest impact on the health of your sample
and fluorophore(s). Increasing laser power will yield more signal but it
will also induce negative photo effects such as phototoxicity and
photobleaching that can harm the sample. Lasers also generate
considerable heat when used at higher powers and that may have
unforeseen effects on the sample.
For most lasers on this microscope, we recommend laser power
settings between 1% and 5% to start. It is recommended to start as low
as you can possibly go and still get an image. Then it can be increased
as necessary to help balance image quality.
To discover your fluorophore(s) saturation level (i.e. how much laser power you can use before your fluorophore stops absorbing
additional photons) you can start imaging at a low laser power and gradually increase the power slider until you reach a point
where your image stops getting brighter. Continual increases in laser power will stop yielding more signal. That point is the
practical laser power limit.
