Confocal images were collected on an LSM 510 laser scanning confocal unit (Carl Zeiss, www.zeiss.com) with a 63X 1.4 numerical aperture plan Apochromat and a 40X 1.3 numerical aperture plan Neofluar objective (Carl Zeiss).
Samples were mounted using antifade reagent with DAPI (ProLong Gold; Invitrogen) and observed using a Zeiss Axiovert 200M microscope with an LSM 710 confocal atttachment, using a 63×1.4 numerical aperture Plan Apochromat oil-immersion lens.
For the high-resolution images shown in the Figures, a 63× 1.4 NA (numerical aperture) plan apochromatic oil-immersion objective was used, and the pinhole was set to a diameter corresponding to 1 Airy unit (optical section ∼240 nm) for true confocal images.
Octavious, whose last name appropriately conjures a numerical superhero, plans to keep "The Stacks" going.
Initially, some aspects and general questions will be addressed to elucidate key points of numerical modelling planning to ensure the validity and reliability of the performed computer simulations.
A microscope objective with high numerical aperture (Plan-Neofluar, 100×/1.30 oil, Zeiss) was used to focus the excitation light as well as to collect the fluorescence emission.
Images were taken on a Zeiss LSM 510 attached to a Zeiss Axiovert 100 M with a 63×1.4 numerical-aperture plan Neofluar oil immersion lens (Carl Zeiss, Thornwood, NY) and processed with Adobe Illustrator software.
Images were acquired by sequential scanning using a ×63 (1.4 numerical aperture) plan-apochromat oil objective and the appropriate filter combination.
Images were acquired using a DMIRE2 Leica confocal microscope (Leica) with a 63× oil immersion 1.40 NA (numerical aperture) plan-apochromat oil immersion lens.
An objective (63× Oil 1.25-numerical aperture, Plan-Neofluar, Zeiss) was coupled with immersion oil to the bottom face of glass coverslips.
