[OpenSPIM] sharp widefield image, but out of focus FP emmission

[Jaap van Krugten]

Dear OpenSPIMers,


We are building an openSPIM with a 100x detection objective as only major
adaptation. Our light sheet is nicely shaped and thin, and if we look at
our sample using a white light LED opposite of the detection objective, it
is in focus. However, if we turn on the laser, we can not get the GFP
emission light in focus. This seems odd, as our wide field image *is* in
focus.

We are quite certain the laser is properly aligned, and we did try moving
the light sheet through our sample. Any ideas what else we can try?


Thanks in advance,

Jaap van Krugten



Jaap van Krugten, MSc | PhD candidate | *in vivo*
single-molecule fluorescence microscopy
Physics of Living Systems, Department of Physics and Astronomy | *Vrije*
 Universiteit Amsterdam

+31 (0)20 59 87904 *|* room U-030, W&N
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