SPIM alignment using multiple wavelengths

openspim

#1

Hi there,

Just wondering how people align their SPIM’s when you want to use multiple laser lines? There is quite a bit of chromatic aberration through achromatic doublets therefore the gaussians of different wavelengths are offset some way laterally. Presumably all I can do is a best compromise in the position for the wavelengths we want to use? Has anyone been able to correct this optically or do you not see much chromatic aberration?

Thanks for the advice,

Matt


#2

Hi @matt_pearson,

I used a multiple laser system from VersaLase (laser lines are pre aligned)
In my experience the chromatic aberration effect in small embryos (180 um) had no significant effect on image quality. But this might depend on your question/sample. You can see it e.g. by taking a closer look at beads.

Cheers,

Johannes