[OpenSPIM] beam sheet thickness and co-axis of double illumination paths

[Luke Stuyvenberg]

Hi Zuomin Zhao,

On 05/06/14, Zuomin Zhao wrote:
> The light sheet thickness of Our SPIM microscope is estimated about 50 micrometers, during adjustment. I remember from OpenSPIM papers, the sheet thickness should be thinner than several micrometer. Could you tell me how to solve it?



Did you completely align the system? Please try adjusting the distance between the first two lenses (the ones on the long rail). The instructions at http://openspim.org/Advanced_Laser_Alignment might also help.


On 05/06/14, Zuomin Zhao wrote:
> We used two illumination beams from opposite direction, how can align the two sheets? how large usually the malposition between the two sheets? Thank you.



This is quite difficult. Perhaps if a sample is mounted in the system, blocking one laser and then the other can show you where the two sheets are, relative to each other? I don't know the usual misalignment; we've only tried using two beams once before.


Good luck; hope this helps!
Luke Stuyvenberg


On 05/06/14, Zuomin Zhao  wrote:
> 
> 
> 
> 
> Hi,
> 
> 
> 
> The light sheet thickness of Our SPIM microscope is estimated about 50 micrometers, during adjustment. I remember from OpenSPIM papers, the sheet thickness should be thinner than several micrometer. Could you tell me how to solve it?
> 
> 
> 
> We used two illumination beams from opposite direction, how can align the two sheets? how large usually the malposition between the two sheets? Thank you.
> 
> 
> 
> Best regards,
> 
> 
> 
> Zuomin Zhao