Evaluation of Slowfade Diamond as a buffer for STORM microscopy.

Area of Science:

• Optical microscopy
• Super-resolution microscopy
• Biophysical imaging

Background:

• Super-resolution Optical fluctuation Imaging (STORM) microscopy requires specific buffer conditions for optimal performance.
• Commercial mounting media are often evaluated for their compatibility with various microscopy techniques and dyes.

Purpose of the Study:

• To evaluate the suitability of Slowfade diamond mounting medium as a buffer for STORM microscopy.
• To determine the compatibility of Slowfade diamond with different fluorescent dyes commonly used in STORM imaging.
• To assess the long-term stability of samples mounted in Slowfade diamond for STORM imaging.

Main Methods:

• STORM microscopy was performed using samples mounted in Slowfade diamond.
• The performance of Slowfade diamond was tested with both far-red (Alexa Fluor 647) and green-excited (Alexa Fluor 532, Alexa Fluor 555, CF 568) fluorescent dyes.
• Sample stability was assessed over several months when stored under refrigeration.

Main Results:

• Slowfade diamond did not support STORM imaging with far-red dyes like Alexa Fluor 647.
• Excellent STORM imaging performance was achieved with green-excited dyes, including Alexa Fluor 532, Alexa Fluor 555, and CF 568.
• Samples remained suitable for STORM imaging for several months when stored refrigerated after mounting.

Conclusions:

• Slowfade diamond is a viable mounting medium for STORM microscopy when using green-excited dyes.
• The medium provides a convenient method for long-term sample preservation and storage of calibration standards for STORM imaging.
• This finding is particularly beneficial for imaging facilities offering metrology and teaching services.