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    • Multiplexed Protein Map analysis
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Welcome to cellatlas.org

This website has been created by the Pelkmans Lab (University of Zurich) to provide the experimental protocols and computational tools to perfrom Iterative Indirect Immunofluorescence Imaging (4i) and to quantify highly multiplexed images with Multiplexed Protein Maps (MPP) analysis.
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To learn how to perform a 4i experiment yourself, or how to construct Multiplexed Protein Maps click on the individual icons below.

4i technology
step-by-step protocol & material list

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Multiplexed Protein Map analysis
MatLab and R code

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4i is a simple, robust and no-degrading protocol for multiplexed indirect immunofluorescence (IF) targeting at least 40 antigens in the same cell with the use of off-the- shelf antibodies. Multiplexing is achieved by iterative cycles of IF and antibody elution. Whilst antibodies are removed efficiently from the sample, epitopes are preserved and virtually unchanged over multiple elution rounds at the level of organelle structure and cellular heterogeneity.

MCU represent areas in cells, which are characterized by a common but unique multiplexed pixel profiles. MCU analysis identifies 
It is worth noting that the such maps are created fully unsupervised and and are independent of the specific size, shape, or orientation of an individual cell or its intracellular organelles.

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