Related Experiment Video
Updated: May 27, 2025
Test Samples for Optimizing STORM Super-Resolution Microscopy
Published on: September 6, 2013
Correlation-based nanometric localization using lattice SIM2 and dSTORM in PML nuclear bodies validated by
Corentin Rousset1, Rémi Neplaz1, Pelin Catal2
1Université Claude Bernard Lyon 1, Laboratoire Physiopathologie et Génétique du Neurone et du Muscle-Institut NeuroMyoGène, Team « Chromatin dynamics, nuclear domains, virus », PGNM-INMG, UCBL - CNRS UMR5261 - INSERM U1315, LAbEx Dev2CAN, Faculté de Médecine et de Pharmacie, F69008, Lyon, France.
Lattice SIM² microscopy effectively visualizes Promyelocytic Leukemia Nuclear Bodies (PML NBs) and their ring-like structures. This advanced technique correlates well with super-resolution microscopy, enhancing 3D imaging capabilities.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biophysics
Background:
- Promyelocytic Leukemia Nuclear Bodies (PML NBs) are crucial nuclear structures involved in various cellular processes.
- Super-resolution microscopy techniques are essential for resolving nanoscale details within cells.
- Lattice di-functionalized 3D structural illumination microscopy (SIM²) offers enhanced resolution over conventional microscopy.
Purpose of the Study:
- To evaluate the performance of Lattice SIM² for imaging PML NBs.
- To compare Lattice SIM² with its predecessor and other super-resolution methods like dSTORM.
- To optimize Lattice SIM² parameters for improved 3D nanoscopic correlation.
Main Methods:
- Application of Lattice SIM² to biological samples (PML NBs) and calibration beads.
- Comparison of Lattice SIM² performance against its predecessor.
- Comparative analysis with direct stochastic optical reconstruction microscopy (dSTORM).
- Utilizing Eternity and Eternity-SIM imaging buffers for 2D and 3D nanoscopic correlation.
Main Results:
- Optimized Lattice SIM² parameters successfully revealed the distinct ring-like morphology of PML NBs.
- A strong correlation was observed between blinking events in dSTORM and the SIM² mask.
- The Eternity-SIM buffer, with its higher refractive index, demonstrated improved 3D correlation for SIM² imaging.
Conclusions:
- Lattice SIM² is a valuable tool for high-resolution imaging of PML NBs, revealing their detailed morphology.
- The study provides insights into the correlation between SIM² imaging and blinking artifacts in other super-resolution techniques.
- Optimized imaging buffers enhance the capabilities of Lattice SIM² for advanced 3D nanoscopic analysis.
