4Pi

Leica 4Pi room Be 336c (4Pi336)

4PI
Lasers: Argon (458, 488, 514), Diode (568), HeNe (633), Mai tai tunable multi-photon laser (710-950nm)

Lenses: 2x HCX PL APO 100x glycerol N.A. 1.35 0.22/0.22

Detectors: 4x Spectral PMT's, 2x Avalanche diodes

Main Usage: Super resolution 3D scans

Contact person:  Gert van Cappellen 


 

4Pi Principle


 

 

 

 

 

 

 

 

 



The excitation spot is split in 1 main lobe and 2 side lobes (Multi-photon  excitation).  This smaller main lobes gives the very high resolution.

  

 

 

 

  

 

   Reality, 100 nm bead XZ-image

 


The raw data of the 4Pi microscope will contain the side lobe information (XZ image beta-tubulin).

 

 

 

 

 

 

 


 

After deconvolution (correction of side lobes) a high resolution image remains

 

 

 

 

 

 


 

 

To give a better impression look at the volume rendered hela cell with beta-tubulin staining.

 

 

 

 

 

 


 


Gert van Cappellen, beta-tubulin slides made by Jeffrey van Haren