Simultaneous visualization of DNA loci in single cells by combinatorial multi-color iFISH (Dataset 1: miFISH chr2 Replicate 1)

We designed 16 probes targeting 16 consecutive loci on chromosome 2 using iFISH4U. The 16 probes targetting different loci are spaced with 3 or 20 Mb in order to study relatively small genomic distances and compare them with larger distances. Furthermore, 10 loci are made of a combination of two colours alternating within the locus and 6 loci have only one colour. Replicate 1

Experiment Imaging Method: epifluorescence microscopy
Protocol Name: iFISH protocol

Each dataset consists of multiple fields of view (FOVs) that were acquired with seven different dyes (AF488 labeled as ‘a488’; AT542 as ‘tmr’; AF594 as ‘a594’; AT647N as ‘Cy5’, AF700 as ‘a700’, AF790 as ‘ir800’; and Hoechst 33342 as ‘dapi’) and all the FOVs are provided as separate .tiff files. Each FOV comprises 81–95 focal planes spaced 0.2 μm apart and has a size of 1024x1024 pixels.