Added 29 October 2020:

Using Zeiss Zen Blue, how to set the correct exposures for widefield fluorescence.

Typically images are too bright with too many saturated pixels. This undermines spatial resolution and the ability to quantify relative numbers of molecules. This video explains how to set good exposures.

It assumes excitation intensity fixed at 100% and imagers will only change exposure time. There is no discussion of camera gain, binning, or field size. This video exclusively addresses exposure time and its influence on dynamic range and LUT adjustments to change image appearance without altering the raw data.

Video version YouTube URL (watch at 720 or 1080 resolution) Direct download URL
ad libbed exposure setting video 1
mostly scripted exposure setting video 2


Do you find that your sample bleaches while you are setting up imaging conditions? This video explains how to avoid this.

  YouTube URL (watch at 720 or 1080 resolution) Direct download URL expose samples to less light


Thank you Ipsita Subudhi in Shruti Naik lab at NYU Langone for sample. Imaging performed on Dept Pathology's Zeiss Imager.Z2 with a 20X PlanApoChromat N.A. 0.8 lens, X-Cite 120LED Boost light source, AxioCam 506 mono, and Zen Blue 2.6.


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