Well plate imaging on 3I Marianas

Things to remember when imaging well plates with DefiniteFocus on 3I:

• Focus on sample
• Check on focus window that z-stage position is approximately zero. If not, restart microscope and DefiniteFocus on their control boxes. Remember to start DefiniteFocus before the microscope
• At focus window - XY - home the stage
• make new slide
• Goto special capture - capture multiwell
• select / edit correct well plate format; note that when you click save the previous plate calibration positions are removed; on 96-well use calibration column 11, rightmost edge of 12 cannot be reached
• Set calibration positions: top middle of the well in top corners and middle right on the bottom right (check few positions by selecting and clicking GOTO)
• Select well you want to image
• Export points
• In focus window XY tab goto position 1 and focus
• look through the z-position values in the position list; if any of the differ very much from the others, reset all z-values
• use DefiniteFocus with same offset for all positions and click set parameters
• In image capture set channels, activate multipoint list and autofocus and set imaging field size
• Start imaging