The road to "nearly" perfect images
The optical process of imaging microscopically small objects is far from perfect. Many artifacts are introduced by the optical setup before an actual image is presented to the observer. It is important to realize that the image is in fact only a representation of the object "what you see is not what you have".
Artifacts that are introduced during the imaging process include for example bleaching (a result of fluorophores not being perfectly photo-stableg), distortions due to spherical aberration (samples having no homogeneous refractive index (distorted spherical aberration) or being too photon-dense (non-linear effects), lenses that are not perfect spheres and chromatically corrected (spherical aberration/geometeric distortion), optical mirrors which are not perfectly well-aligned, and stages/objectives that are not completely resistant to temperature fluctuations, which may result in drift and stability issues. See also AcquisitionArtifacts
To aim for "the perfect image" and get the best representation of your object you should:
- optimize sample preparation
- select the right microscope system for your experiment
- calculate the optimal sampling values with the NyquistCalculator to image enough detail of your objects
- use the proper image acquisition conditions
- apply the correct image restoration options
- follow the right guidelines for signal quantification
Your image facility manager is probably the first to contact for advise. Of course, we would be happy to help you also with your quest to obtain reliable and more imaging results. You can contact us at info
