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Daybook Analysis - Version 2.0.0 - October 2024

This analysis only concerns microscopes equipped with a motorized Z stage.

Z-stack imaging is very common for imaging and reconstructing a specimen in 3D

When acquiring Z-stack images, the XY stage may drift, distorting the reconstructed volume of the entire specimen. It is therefore important to measure the stage drift, so that it can be corrected for, and does not introduce bias into the measurement of the specimen trajectory. This can also trigger a maintenance session.

Sources of stage drift during Z-stacking typically come from environmental conditions (such as temperature fluctuations and air flow) or stage malfunctions.

The “stage drift during Z-stacking” analysis enables the measurement of the lateral drift magnitude from the XY stage during Z-stack imaging, to ensure it is within the specifications of the manufacturer or experimentalist.

I. IMAGE ACQUISITION PROCEDURE

The “stage drift during Z-stacking” analysis is associated with the “3D crossing stairs” patterns (Pattern family I - See Figure 1).

Figure 1: Example image of the “3D crossing stairs” pattern, fulfilling the acquisition recommendations.

Figure 1: Example image of the “3D crossing stairs” pattern, fulfilling the acquisition recommendations.

1. ACQUISITION RECOMMENDATIONS

2. IMAGING THE PATTERN

II. IMAGE ANALYSIS PROCEDURE

1. LAUNCHING THE ANALYSIS

2. ANALYSIS SETTINGS