Highly Multiplexed Immunofluorescence Imaging of Mouse Oocytes
Cyprien Noble1, Typhaine Esteves2,3, Adel Al Jord4,5
1Center for Interdisciplinary Research in Biology (CIRB), Collège de France, CNRS, INSERM, Université PSL, Paris, France.
Insights
We developed a cost-effective iterative indirect immunofluorescence imaging (4i) protocol to visualize tens of proteins in single mouse oocytes. This method enhances understanding of cellular biology in challenging samples.
Area of Science:
- Cellular and Molecular Biology
- Biotechnology
- Microscopy
Background:
- Highly multiplexed immunofluorescence imaging is crucial for understanding biological processes at molecular and tissue levels.
- Its application is particularly valuable for scarce and difficult biospecimens, such as mammalian oocytes.
- Existing methods are often expensive and complex, limiting their widespread use.
Purpose of the Study:
- To present a cost-effective and simple protocol for highly multiplexed immunofluorescence imaging.
- To enable the visualization of tens of proteins within a single mouse oocyte.
- To provide an adaptable method for other large, non-adherent cells.
Main Methods:
- Development of an iterative indirect immunofluorescence imaging (4i) protocol.
- Application of the protocol to single mouse oocytes.
- Focus on capturing protein distribution and abundance.
Main Results:
- Successful visualization of tens of proteins in individual mouse oocytes.
- Demonstration of a cost-effective and simplified imaging approach.
- Validation of the protocol's adaptability to other cell types.
Conclusions:
- The iterative indirect immunofluorescence imaging (4i) protocol offers an accessible method for high-content protein analysis in oocytes.
- This technique can advance research in reproductive biology and developmental processes.
- The protocol's adaptability broadens its potential applications in cell biology.
Abstract:
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.
