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Daybook Analysis - Version 1.10.0 - October 2023

The photometric calculator is a tool that allows the computation of the essential photometric metrics to know for quantitative imaging and reproducibility, for four acquisition modes: wide-field, confocal laser scanning, confocal spinning disk, and two-photon laser scanning microscopy. The computed photometric metrics are peak power, irradiance, dose, and photon flux.

The photometric calculator also aims at being an instructive tool to help understand how the illumination settings (average power, wavelength, objective, etc.) influence the photometric metrics, and how the different acquisition modes (wide-field, confocal, etc.) can give rise to photometric metrics that can differ by several orders of magnitude for the same input parameters.

I. MICROSCOPY ACQUISITION MODES

A sketch of the four different microscopy acquisition modes in the “Photometric calculator” is shown in Figure 1, and their main intrinsic differences are summarized in Table 1.

Figure 1: Sketches of the different microscopy acquisition modes. From left to right: wide-field, confocal laser scanning, confocal spinning disk, and two-photon laser scanning microscope.

Figure 1: Sketches of the different microscopy acquisition modes. From left to right: wide-field, confocal laser scanning, confocal spinning disk, and two-photon laser scanning microscope.

Table 1: Main intrinsic differences between the acquisition modes. A camera is an example of an array detector. A photomultiplier tube is an example of a point detector.

Table 1: Main intrinsic differences between the acquisition modes. A camera is an example of an array detector. A photomultiplier tube is an example of a point detector.

Because of these intrinsic differences, the photometric metrics calculated from the same input parameters (illumination power, objective, wavelength) will be highly distinct.

II. INPUT PARAMETERS

1. LIGHT SOURCE FEATURES