Essentials II – Essentials of confocal microscopy (online)

Organizer: Manuel Gunkel, Aliaksandr Halavatyi and Marko Lampe

 Registration for EMBL staff only 

The course is covering the essential knowledge needed to conduct a confocal microscopy-based experiments. Along with that, we discuss advanced topics of microscopy and fluorescence imaging.The prerequisite for attending this course is the knowledge covered in Essentials I course. The microscopy knowledge covered in this is mandatory for introductions into ALMF Confocal-microscopes.

Please be aware, that this course mainly cover practical aspects with only a limited amount of theory. Therefore, we strongly suggest to watch the following recommended tutorials in advance.

Course outline

Practical point scanning confocal – part 1:

1. Why/when one needs optical sectioning
2. Lightpath
3. Why need to scan
4. Confocal principle
5. Pinhole size
6. 3D data acquisition with confocals

Practical point scanning confocal – part 2:

1. Lateral and axial resolution
2. Selecting pixel size
3. Choosing an objective
4. Saturation and offset
5. SNR Averaging/Accumulation, Voltage, laser power

Practical point scanning confocal – part 3:

1. How to achieve higher frame rates
2. 3D proer stack configuration
3. XZ scanning
4. Refractive index and spherical aberrations
5. Correcting for z-distortion
6. Minimizing spherical aberrations

Practical point scanning confocal – part 4:

1. Multicolor imaging – use of sequences and tracks
2. Multicolor imaging – configuration of detectors
3. Chromatic aberrations
4. Point scanning multicolor imaging – spectral detectors
5. Getting transmitted light image
6. Sample types

Q&A session

Bookings