🟤Microscopy Tutorials

Essential Microscopy Tutorials and Protocols

Essential Microscopy Tutorials and Protocols Community-Curated by GerBI-GMB

Introduction

The GerBI-GMB (German BioImaging - Society for Microscopy and Image Analysis) community has curated a valuable collection of short educational videos and detailed protocols. These resources guide best practices in microscopy, helping users maintain, calibrate, and optimize their instruments for high-quality imaging. Below is a summary of key tutorials, direct access to each video, and accompanying protocols.

1. How to Measure a Point Spread Function (PSF)

Purpose: Determine the lateral and axial resolution of your microscope.

• Video: How to Measure a PSF – Nico Stuurmann (iBiology)

• Protocol: QUAREP-LiMi Protocol for PSF Measurement

This video includes bead slide preparation and optimal acquisition settings. QUAREP-LiMi provides a detailed written protocol that includes image acquisition and analysis steps.

2. How to Clean Objective Lenses and Filters

Purpose: Maintain optical performance by safely removing dust and residues.

• Video: Cleaning Lenses and Filters – Kurt Thorn (iBiology)

Learn to inspect and clean optical components without damaging their surfaces, ensuring sustained image clarity and protection of expensive parts.

3. How to Find the Focal Plane

Purpose: Accurately focus on the specimen without harming it.

• Video: Finding the Focal Plane – Ron Vale (iBiology)

This tutorial provides tips on achieving focus smoothly, minimizing risks to both sample and objective lenses.

4. How to Set Up Koehler Illumination

Purpose: Achieve even illumination across the specimen’s focal plane.

• Video: Koehler Illumination Setup – Ron Vale (iBiology)

Understand the principle of Koehler illumination and follow a step-by-step alignment of lamp and condenser.

5. Live Cell Imaging and Environmental Control

Purpose: Ensure viability and reproducibility in live-cell imaging.

• Video: Environmental Control in Live Cell Imaging – Kurt Thorn (iBiology)

Explore strategies for maintaining temperature, humidity, and CO₂ levels, ranging from basic to advanced setups.

6. How to Minimize Photobleaching and Phototoxicity

Purpose: Protect live samples from light-induced damage during imaging.

• Video: Reducing Photobleaching and Phototoxicity – Ron Vale (iBiology)

Learn about minimizing light exposure and reactive oxygen species in fluorescence imaging experiments.

7. How to Calibrate a Camera

Purpose: Optimize camera performance through quantitative calibration.

• Video: Camera Calibration – Nico Stuurmann (iBiology)

This video walks you through:

• Capturing dark images

• Estimating readout noise and dark current

• Generating flat field images

• Measuring photon conversion factor and full well capacity

Disclaimer

This compilation is for educational and technical reference. Videos are hosted by iBiology and protocols are provided by QUAREP-LiMi. GerBI-GMB does not assume responsibility for implementation outcomes.