Core Facilities
USF Health - College of Medicine

 
      


Molecular Probes Storm 860 Imager

Phosphoimager is able to scan intensifying screens or detect fluorescence at two different wavelengths.
The Molecular Dynamics Storm 860 Phosphoimager is able to quantify radioactive gels. The standard phosphor screen is designed for commonly used isotopes such as 32P, 33P, 125I, 35S, and 14C. Since the Phosphoimager is much more sensitive than ordinary x-ray film, exposure times can be reduced by up to ten-fold. In addition, signal quantitation over several orders of magnitude of both weak and strong signals can be further adjusted on the same blot using the ImageQuant software.

The Molecular Dynamics Storm 860 Phosphoimager can also detect direct fluorescence, and chemifluorescence using blue and red channels. Therefore, the instrument can be used for: chemi-fluorescence/blue-excited scans (450 nm excitation) and red-excited fluorescence scans (635 nm excitation laser). Applications using this capability include quantitation of: fluorescence stained nucleic acid gels (EtBr; SYBR green), protein gels stained with SYPRO Orange and SYPRO Red, and chemi-fluorescence detection for Western Blots using Amersham’s ECL Plus substrate.

Storm 860 Procedure

1. Open Scanner Control software.
2. Select Scanner type according to your own experimental needs. For Phosphoimager, select the “STORM 860 – Phosphor Screen”, press “OK”.
3. Outline the detecting area.
4. Save the settings in a file on desktop. Remember to transfer the file to your own disk and delete your file from desktop.
5. Click “Scan”. The scanner will automatically start the scanning.
6. Once finished scanning, open the gel document with ImageQuant 5.2 software.
7. To adjust the brightness of the image, go to “View” and click the “Gray/Color Adjust”. Pixel density represents quantity.
8. Go to “Object”, click “Select” then choose the appropriate shape to cover the area you want to analyze.
9. Go to “Analysis”, click “Volume Report” and highlight all the areas you want to analyze, the program will show the values of all the areas selected.
10. Data can be imported to Microsoft Excel for further analysis.