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Updated: Oct 17, 2025
Optical Clearing and Imaging of Immunolabeled Kidney Tissue
Published on: July 22, 2019
Highly multiplexed immunofluorescence of the human kidney using co-detection by indexing
Elizabeth K Neumann1, Nathan Heath Patterson1, Emilio S Rivera1
1Department of Biochemistry, Vanderbilt University, Nashville, Tennessee, USA; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee, USA.
Insights
This study showcases co-detection by indexing (CODEX) multiplexed immunofluorescence for mapping 23 human kidney cell types. This technique reveals cellular differences in healthy versus diabetic kidney tissues, aiding anatomical atlases.
Area of Science:
- Renal cell biology
- Immunofluorescence techniques
- Tissue architecture analysis
Background:
- The human kidney comprises diverse cell types crucial for organ function.
- Accurate cell identification is vital for understanding tissue architecture and microenvironments in health and disease.
- Multiplexed imaging techniques are needed to visualize multiple cell types simultaneously within kidney tissue.
Purpose of the Study:
- To demonstrate the utility of co-detection by indexing (CODEX) multiplexed immunofluorescence for visualizing 23 antigens in the human kidney.
- To assess the flexibility and applicability of CODEX for studying human kidney tissues using custom and commercial antibodies.
- To compare cellular differences between healthy and diabetic nephropathy kidney tissues using CODEX.
Main Methods:
- Co-detection by indexing (CODEX) multiplexed immunofluorescence was employed to visualize 23 antigens in human kidney tissue sections.
- Antibodies for 19 antigens were conjugated in-house, highlighting the adaptability of the CODEX approach.
- A pilot study compared fresh frozen and formalin-fixed paraffin-embedded tissues from healthy and diabetic nephropathy kidneys.
Main Results:
- CODEX successfully visualized major kidney cell types and substructures, including collecting ducts, glomeruli, and thick ascending limb, within a single tissue section.
- Significant cellular differences between healthy and diabetic nephropathy kidney tissues were identified, particularly in cells expressing aquaporin 1, cytokeratin 7, and α-smooth muscle actin.
- The study demonstrated the feasibility of using in-house conjugated antibodies with CODEX for human kidney research.
Conclusions:
- CODEX multiplexed immunofluorescence is a powerful tool for surveying the cellular diversity of the human kidney.
- This technique holds significant potential for applications in pathology, histology, and the development of comprehensive anatomical atlases.
- CODEX enables detailed cellular analysis, facilitating the understanding of kidney diseases like diabetic nephropathy.
Abstract:
The human kidney is composed of many cell types that vary in their abundance and distribution from normal to diseased organ. As these cell types perform unique and essential functions, it is important to confidently label each within a single tissue to accurately assess tissue architecture and microenvironments. Towards this goal, we demonstrate the use of co-detection by indexing (CODEX) multiplexed immunofluorescence for visualizing 23 antigens within the human kidney. Using CODEX, many of the major cell types and substructures, such as collecting ducts, glomeruli, and thick ascending limb, were visualized within a single tissue section. Of these antibodies, 19 were conjugated in-house, demonstrating the flexibility and utility of this approach for studying the human kidney using custom and commercially available antibodies. We performed a pilot study that compared both fresh frozen and formalin-fixed paraffin-embedded healthy non-neoplastic and diabetic nephropathy kidney tissues. The largest cellular differences between the two groups was observed in cells labeled with aquaporin 1, cytokeratin 7, and α-smooth muscle actin. Thus, our data show the power of CODEX multiplexed immunofluorescence for surveying the cellular diversity of the human kidney and the potential for applications within pathology, histology, and building anatomical atlases.
