Ve. 1225 1

1. Prep the workspace

• Gather the kit: get the slide out of the storage box, have the latest version of Daybook 4 software installed on the workstation beforehand (internet connection required). Have your Daybook 4 account created, and license entered (the license code ships with the slide).  https://argolight.com/products/daybook-4/
• Clean the glass surface and your objective with a lens tissue lightly soaked with a cleaning solvent (ethanol, isopropyl alcohol...)
• Power on the microscope and warm up the light source for at least one hour (especially for confocal).

2. Load & align the slide

• Select a low‑magnification objective*(5x recommended)* first so you can locate the patterns quickly without hunting with a high‑NA lens. Use the DAPI channel (350‑405 nm). The pattern shines brighter in DAPI.

• Place the Argolight slide on the stage, face‑up (for upright microscopes).

• Turn on the camera. Select the auto-exposure option in your acquisition software.

• •Coarse‑align the slide center with the camera’s field of view using the XY and Z translation knobs.

• Select the pattern of interest for your quality control (the Gradually Spaced Lines for Lateral resolution analysis). Go to the center of the pattern of interest.

  Tips: The line spacings run from 0 nm to ~390 nm, perfect for probing the smallest resolvable feature of the imaging system.

• Then, navigate Z. Fine-align the focal plane.

E.g. of Gradually Spaced Lines (GSL) pattern from an Argo-SIM slide.

3. Switch to the relevant acquisition settings

We’ll now switch to the acquisition settings of interest.

1. Add the immersion medium relevant to the objective on the piece of glass.

2. Switch to the working objective (e.g., 60× oil) if you want higher resolution.

3. Switch to your channel of interest, usually GFP (488nm).

4. Fine-align the focal plane.

Tip: For confocals, especially, avoid zooming in as much as possible (to avoid risks)