[ALMF]: Essentials II – Essentials of confocal microscopy (online)

almf   2021-01-28   Comments Off on [ALMF]: Essentials II – Essentials of confocal microscopy (online)

Date(s) - 2021-01-28 - 2021-01-29
09:00 CET - 12:00 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.

Teaching material:

  1. Basics of confocal microscopy
  2. Nikon Microscopy University – Introductory Confocal Concepts
  3. Bit depth and resolution in an image
  4. Choosing the right microscopy technique

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

Practical point scanning confocal – part 2 (stained cells – will use mainly one channel)

  1. Lateral and axial resolution
  2. Selecting pixel size
  3. Saturation and offset
  4. SNR Averaging/Accumulation, Voltage, laser power
  5. How to achieve higher frame rates (e.g. time lapse): scan speeds, number of pixels, bi-directional scan

Practical point scanning confocal – part 3

  1. 3D data acquisitions with confocals – proper stack configuration
  2. Multicolor imaging – use of sequences and tracks
  3. Multicolor imaging – configuration of detectors
  4. Point scanning multicolor imaging – spectral detectors
  5. Getting transmitted light image

Q&A and discussion round – alternatives to point scanning confocal for optical sectioning

  1. TIRF
  2. Spinning disk
  3. Multiphoton
  4. AryScan
  5. SPIM


This event is fully booked.