Detailed protocols for monitoring acidification of endocytosed probes using flow cytometry may be found in Sheree Lynn Rybak and Robert F. Murphy (1997) "Measurement of Ligand Acidification Kinetics for Adherent and Non-Adherent Cells" in In Living Color -- Protocols in Flow Cytometry and Cell Sorting (Rochelle Diamond and Susan L. DeMaggio, eds.), R.G. Landes Co., in press. The following spreadsheets (in different formats) are referred to in that chapter and are provided to illustrate how dual-fluorescence measurements are converted to estimates of pH:

• Spreadsheet for averaging and normalizing calibration curve of FITC/Cy5 vs. pH (Protocol 4a)
• Excel 5.0
• Lotus 1-2-3 (WK1)
• Symbolic Link format (SYLK) [charts not included]
• Spreadsheet for calculating compartment pH for cells labeled with FITC- and Cy5-dextran (Protocol 5a)
• Excel 5.0
• Symbolic Link format (SYLK) [charts not included]
• Spreadsheet for calculating compartment pH for cells labeled with FITC- and Cy5-transferrin (Protocol 5b)
• Excel 5.0
• Symbolic Link format (SYLK) [charts not included]

Last Updated: 01 Dec 2004

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