Date(s) - 2021-02-09 - 2021-02-10
14:00 CET - 17:00 CET
Registration for EMBL staff only
The course is covering the essential knowledge needed to conduct a confocal microscopy-based experiments. 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.
- Basics of confocal microscopy
- Nikon Microscopy University – Introductory Confocal Concepts
- Bit depth and resolution in an image
- Choosing the right microscopy technique
Practical point scanning confocal – part 1:
- Why/when one needs optical sectioning
- Why need to scan
- Confocal principle
- Pinhole size
Practical point scanning confocal – part 2 (stained cells – will use mainly one channel)
- Lateral and axial resolution
- Selecting pixel size
- Saturation and offset
- SNR Averaging/Accumulation, Voltage, laser power
- How to achieve higher frame rates (e.g. time lapse): scan speeds, number of pixels, bi-directional scan
Practical point scanning confocal – part 3
- 3D data acquisitions with confocals – proper stack configuration
- Multicolor imaging – use of sequences and tracks
- Multicolor imaging – configuration of detectors
- Point scanning multicolor imaging – spectral detectors
- Getting transmitted light image
Q&A and discussion round – alternatives to point scanning confocal for optical sectioning
- Spinning disk
This event is fully booked.