Content Comparison

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In the image below I have sealed the middle two wells by sliding the inverted lid across them.

Focusing on the specimen

Adjust the focus (Z dimension) for both microscopes to the following positions to find the approximate focus for the middle two wells of the ibidi chamber.

• Zeiss Elyra PS.1: 4.622
• ONI Nanoimager: -240.0

Coloc-Tesseler

Levet F, Julien G, Galland R, Butler C, Beghin A, Chazeau A, Hoess P, Ries J, Giannone G, Sibarita J-B. A tessellation-based colocalization analysis approach for single-molecule localization microscopy. Nature Communications, 10, 2379. 2019.

Link to Github downloads.

Colocalisation measurements in Coloc-Tesseler

Spearman Rank Correlation Coefficient

Like the Pearson Correlation Coefficient, this measures the degree to which the values of individual measurements of two channels change together either positively (correlated) or negatively (anti-correlated). A value of +1 indicates 100% correlation and −1 indicates 100% anti-correlation. A value of 0 indicates random correlation. Unlike the Pearson coefficient, the channel measurements do not have to be linearly related to give a correlation close to 1.

Manders Coefficients

The Manders Coefficients show the colocalised values from each channel as a ratio of the total fluorescence for that channel, so there is one coefficient for each channel.

Manders EMM, Verbeek FJ, Aten JA. Measurement of co-localization of objects in dual-colour confocal images. Journal of Microscopy, 169(3), 375, 1993.