Calculate Absorbance from Image Studio™ Software Quantification Values

These instructions explain how to quantify wells in Image Studio and calculate absorbance from those quantification values using spreadsheet software.

  1. If you need to import images from Empiria Studio® Software or another instance of Image Studio, use the LI-COR Zip Files import option.

    The following instructions are for images imported using the LI-COR Zip Files option.

    Do not use the Third-Party import option for this procedure.

  2. Select the image you need to analyze in the Images table.

  3. Apply a Plate Analysis.

    Plate option on the Analysis Type dropdown menu.
    Figure 6. On the Analysis ribbon, choose Plate from the Type menu to apply the Plate Analysis.

  4. Reduce the size of the wells from the Plate Analysis template to fit the inner area of the well.

    In Empiria Studio, this is reduced to 75% of the diameter for circular well or 75% of the width/height of square wells.

    Applied Plate Analysis template with Plate Analysis ribbon shown
    Figure 7. Click the Larger Larger button and Smaller Smaller button buttons to adjust the well size.

  5. In the Plate Wells table, add the Well Name and Max columns.

    Plate Wells table including Well Name and Max value columns
    Figure 8. If Well Name and Max columns are not shown in the Plate Wells table, click Icon for columns button used to customize which columns are shown in tables. Columns to view the dialog for adding columns.

  6. Click Report Report button above table above and to the right of the Plate Wells table and choose one of the options to exporting data to a spreadsheet file or a CSV file that can be opened in a spreadsheet program.

    Report menu open showing export options
    Figure 9. The Report options include launching a spreadsheet directly or saving a .xls file.

  7. In the spreadsheet, create a new cell that will hold the value of the average of the Background or Blank wells.

  8. Define a column that will contain the Absorbance values.

  9. Use the following formula in the Absorbance column to calculate absorbance:

    LOG10(Average of Background Max Values / Sample Max Value)