Highly multiplexed immunofluorescence imaging methods advanced our understanding of biology across scales, from tissues down to molecules. By enabling the visualization of tens of proteins in a single sample, highly multiplexed imaging is especially relevant for scarce and challenging biospecimens like mammalian oocytes. However, most methods remain relatively costly and complex to implement. Here, we provide a cost-effective simple protocol, based on iterative indirect immunofluorescence imaging (4i), allowing to capture the distribution and abundance of tens of proteins in a single mouse oocyte. Our approach is adaptable to other mammalian oocytes or analogously large non-adherent cells like the early embryo.