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Improving resolving ability of expansion microscopy by varying crosslinker concentration
Yanlei Hu1, Ximing Chu1, Ting-Ting Chen1
1State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China. xiachu@hnu.edu.cn.
Chemical Communications (Cambridge, England)
|March 14, 2020
Summary
Expansion microscopy (ExM) was optimized using a specific N,N'-methylenebisacrylamide (MBAA) concentration. This enhanced ExM method achieved a 5.7x expansion factor and ~50 nm resolution with standard confocal microscopy.
Area of Science:
- Biotechnology
- Microscopy techniques
- Cell biology
Background:
- Expansion microscopy (ExM) enables super-resolution imaging but requires optimization for specific applications.
- Different crosslinker concentrations can significantly impact ExM performance and expansion factors.
Purpose of the Study:
- To systematically investigate the effect of N,N -methylenebisacrylamide (MBAA) concentration on ExM performance.
- To optimize ExM for enhanced resolution and expansion factor using a common confocal microscope.
Main Methods:
- Systematic investigation of ExM with varying MBAA concentrations.
- Modification of ExM protocol using 0.06% MBAA (0.06%-MBAA ExM).
- Evaluation of lateral resolution using standard confocal microscopy.
Main Results:
- Achieved an expansion factor of 5.7 with 0.06%-MBAA ExM.
- Obtained a lateral resolution of approximately 50 nm.
- Demonstrated the efficacy of optimized ExM with a conventional confocal microscope.
Conclusions:
- Optimized ExM using 0.06% MBAA significantly enhances expansion factor and resolution.
- This improved ExM protocol is compatible with standard confocal microscopy, making super-resolution accessible.
- The findings provide a refined ExM method for detailed cellular imaging.
