Confocal calibration & performance assesment

Posted by F Javier Diez Guerra on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Confocal-calibration-performance-assesment-tp7587224.html

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Dear confocalists,

I would like to know about the preferred methods used by facility
managers to regularly asses the performance of their confocal (and
wide-field) microscopes. In other words, which are the most used and
reliable methods to check different features of confocal and wide-field
microscope peformance?.

Particularly, we'd like to check:

- illumination uniformity across FOV, now we use a slide coated with
fluorescent secondary antibody.

- XYZ Chromatic aberration (exciting 405, 488, 561 and 640 nm), now we
use 1um beads that can be excited with all laser lines.

- XYZ Resolution, we use smaller beads (0,17um) to build and analyze
PSFs (for each wavelength)

- Laser power and stability: for power, we use a power meter from
Newport placed at the objective exit, for stability, we capture long
time-series in reflexion mode with an empty preparation.

- We do not know how to check detector (PMT, GAsP, Hybrid, etc)
sensitivity and SNR.

Thanks for your input.


--
Fco. Javier Diez-Guerra, PhD

Profesor Titular UAM
Servicio de Microscopía Confocal
Centro de Biologia Molecular Severo Ochoa
C/ Nicolás Cabrera, 1
Campus de Cantoblanco
28049 Madrid
SPAIN

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