Pollen confocal image

[luckyclover]

Hi all,

Just for your interest, this is a confocal stack of some pollen grains found in my honey. The stack has been average projected to form the 2D image after deconvolution in Image J.

The pollen was then artificially coloured using Photoshop.

LC

 

[itma]

Did you do the focus stacking in Photoshop as well? (Or what worked for you?)

And did you just tweak the focus manually?


A decent focus-stacking "how-to" would make an excellent sticky thread ...

 

[luckyclover]

Hi Itma,

No to both questions. In the confocal microscope out of focus light is blocked with a pinhole. I set the z dimensions and how many steps or focus planes and how thick they should be, then it acquires all the slices. In Image J I deconvolved the stack to increase the resolution.
All the optical sections were then combined to form an average projection. With confocal imaging you look in 3 dimensions at your specimen if you want.

Hope that makes sense

 

[luckyclover]

To adjust the z height I am using a motorised stage. Its a 3/4 million pound scope!

 

[itma]

To adjust the z height I am using a motorised stage. Its a 3/4 million pound scope!

I think I'll wait until it turns up in the Sales!

 

[REDWOOD]

Lovely image way way beyond me and my budget