Daybook Analysis - Version 1.10.0 - October 2023
This analysis only concerns microscopes equipped with a motorized Z stage.
Confocal fluorescence microscopes enable 3D imaging, thanks to their ability to remove the out-of-focus light. 3D imaging is achieved by reconstructing a Z-stack of images. The accuracy of this 3D reconstruction may be affected by various issues. Sources of inaccuracy include: the Z-stage, a poorly adjusted objective correction collar for coverslip thickness, an incorrect refractive index of the immersion medium, and environmental conditions, such as temperature fluctuations and air flow.
The “accuracy of 3D reconstruction” analysis allows the control of confocal fluorescence microscopes’ ability to correctly acquire Z-stacks of images, so that their reconstruction into 3D images is accurate, i.e. representative of reality.
The “accuracy of 3D reconstruction” analysis is associated with the “sphere” pattern (Pattern family G - See Figure 1).
Figure 1: Example image of the “sphere” pattern, fulfilling the acquisition recommendations.