OptiGrid Setup

Requires a separate license

Before you use the OptiGrid, set the operating parameters with the OptiGrid Setup dialog box.

Grid Focus is different for each objective and filter you use.

Use the slider to manually focus the OptiGrid in the light path until you find a position where the grid lines are sharpest in the live image.

AF: Automatic Focus

The software starts the automatic focus procedure and stops when it determines the optimal focus.

Grid Focus Presets

Presets are required for multichannel acquisitions.

For best results, identify and save the best grid focus for each objective to be used as part of the initial setup. Loading presets is more efficient than searching for the best focus each time you change the objective.

  1. Click the Presets icon .
  2. Click Edit Presets.
  3. Enter a name and save.
  1. Click the Presets icon .
  2. Click the name of the desired preset.
  3. The parameters are loaded.

OptiGrid Speed—Refers to length of the delay before the OptiGrid settles after grid movement. We recommend you start with the Fast setting and adjust from there.

Section Quality—The higher the quality, the more images the OptiGrid acquires. The default is Fast Acquire.

Apply Fade Correction—Useful for fast-fading dyes or unstable light sources.

  • Click Connect to connect the OptiGrid to the controller. The OptiGrid system will automatically try to connect if powered on and connected to the computer and microscope.
  • Use Calibrate to calibrate the OptiGrid with your hardware. Calibration is typically only needed once for your entire system. Recalibration isn't necessary when changing objectives or lenses.
  1. Place the calibration slide in the microscope.
  2. Focus on the top of the tape at a ragged tape edge.
  3. Move the edge out of view.
  4. Click Calibrate. The system tells you if the OptiGrid failed or succeeded calibration.

Calibration Slides

You can use the Qioptiq-supplied calibration slide or create your own.

Qioptiq supplied calibration slide

QLI provides a calibration slide along with all OptiGrids. It is made up of a thick layer of fluorescent dye for use under DAPI, FITC, and TRITC illumination. Its disadvantage is a reduction in grid contrast (due to the thickness of the dye). It can be prone to photo-bleaching.

User-created calibration slide

If you don't have the calibration slide provided by Qioptiq, create your own. The slide will have a very thin layer of fluorescent dye, which allows grid contrast to be maximized. It's not affected by photo-bleaching. Its disadvantage is that the dye will clump over time; as a result, its lifetime is limited and it can only be used with FITC illumination.

To create a calibration slide for the OptiGrid

  1. Have a fluorescent highlight marker, a microscope slide, and a coverslip available.
  2. Rub the fluorescent marker onto the slide.
  3. Place a cover slip on top of the slide.