4Pi-confocal microscopy provides three-dimensional images of the microtubule network with 100- to 150-nm resolution.
We show the applicability of 4Pi-confocal microscopy to three-dimensional imaging of the microtubule network in a fixed mouse fibroblast cell. Comparison with two-photon confocal resolution reveals a fourfold better axial resolution in the 4Pi-confocal case. By combining 4Pi-confocal microscopy with Richardson-Lucy image restoration a further resolution increase is achieved. Featuring a three-dimensional resolution in the range 100-150 nm, the 4Pi-confocal (restored) images are intrinsically more detailed than their confocal counterparts. Our images constitute what to our knowledge are the best-resolved three-dimensional images of entangled cellular microtubules obtained with light to date.
Nagorni
M
M
High Resolution Optical Microscopy Group, Max-Planck-Institute for Biophysical Chemistry, Göttingen, D-37070, Germany.
Hell
S W
SW
D016475:3T3 Cells / Q000648:ultrastructure*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal / Q000379:methods*; D008870:Microtubules / Q000648:ultrastructure*; D014404:Tubulin / Q000276:immunology
D000906:Antibodies; D014404:Tubulin