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 (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, etc.).
- 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. Set the DAPI ( 405 nm ) or GFP ( 488 nm ) channel.
- 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 ( $4 \times 4$ intensity gradation or $2 \times 16$ gradation). Go to the center of the pattern of interest.
- Focus into the glass until the fluorescent patterns pop into view.
.png)
E.g. of a 4x4 intensity gradation pattern from an Argo-HM-slide.
.png)
E.g. of a 2x16 gradation pattern from an Argo-HM-slide.
3- Configure your microscope for imaging
We’ll now switch to the acquisition settings of interest.
- Add the immersion medium relevant to the objective on the piece of glass.
- Switch to the working objective.
- Switch to your channel of interest, usually GFP $(488 \mathrm{~nm})$.
- Adjust the focus
Tip: For confocal especially, avoid as much as possible to zoom into the pattern (to avoid risks)