Using Contextual Imaging to Quantify Cell Density
15. January 2025
The specific characteristics of individual cells can only offer so much detail during analysis or categorization.
Offering additional reliability to an examination, contextual imaging can provide information besides the cell's phenotype including its positioning, as well as its cellular neighborhood. Instead of focusing on the cell itself, contextual imaging considers the cell within its environment. Contextual imaging includes quantification of cell density, utilizing mapping to understand spatial distributions of cell populations. Moreover, by quantifying cell density, and examining spatial distribution, contextual imaging offers the opportunity to learn about cell subsets, such as T cells. Two techniques that detail cell density are multiplexed immunofluorescence staining, a method that uses sequential staining, fluorescent dyes, and acquisition of the stained tissue section with a slide scanner to create a picture of a tissue's spatial organization, and follow up automated image analysis. Join us as we review these techniques in more detail, considering their application in the examination of esophageal squamous cell carcinoma (ESCC), and the quantification of cell density through contextual imaging.
Materials and Methods
The study utilized an ESCC tissue micro array. This array included 93 cases of ESCC and 87 cases of matched adjacent normal tissue (ANT). It was stained with multiplexed immunofluorescent (mIF) markers for MRPL13, CD8A, and pan-cytokeratin (Pan-CK), among others. Images were acquired using TissueFAXS Spectra system and marker distribution was analysed using StrataQuest analysis software (TissueGnostics). Statistical analyses were conducted to compare the expression levels of MRPL13 in different tissue contexts. Further correlations of MRPL13 with CD8+ T-cell infiltration were explored.
Results
mIF staining provided detailed contextual imaging and allowed for the specific localization and quantification of MRPL13 expression. It was detected in both the stromal and epithelial regions of the ESCC samples. Notably, MRPL13 was found to be significantly elevated in the stromal regions of ESCC tissues compared to ANT. Furthermore, MRPL13 expression was strongly correlated with the infiltration of CD8+ T cells in these regions. These results indicated the immunological role of MRPL13 in the tumor microenvironment.
Automated image analysis facilitated the quantification of cell density by distinguishing between different cell types based on their staining patterns. This approach provided a method for assessing the density and distribution of immune cells in relation to tumor cells within the same tissue section. Therefore, insights were offered that were not readily attainable through traditional histological techniques.
Discussion
The study's use of contextual imaging through multiplexed immunofluorescent staining techniques combined with in-depth image analysis illustrates a significant advancement in how researchers can:
- Quantify cell density
- Assess the cellular architecture of tumors
Contextual imaging provides spatial resolution and the functional status of different cell populations. As a result, the technique can enrich the understanding of tumor biology and the immune environment.
Such detailed analysis is especially critical in the context of personalized medicine, where understanding the microenvironment's complexity can influence treatment decisions. For instance, the observed correlation between MRPL13 expression and CD8+ T-cell infiltration could help tailor immunotherapeutic strategies. This approach could be useful in cancers where immune evasion plays a role in disease progression and treatment resistance.
Furthermore, the findings suggest potential pathways through which MRPL13 may modulate immune responses, presenting new targets for therapeutic intervention. Future studies could explore the mechanistic basis of MRPL13's role in immune regulation. Related research could lead to novel approaches to enhance the efficacy of therapies or develop new therapeutic modalities.
Final Thoughts on Contextual Imaging
Contextual imaging, facilitated by mIF staining and sophisticated image analysis, represents a powerful tool for the quantification of cell density in histological samples. Ultimately, this contextual imaging technique enhances the accuracy of measurements and provides deeper insights into the cellular compositions and interactions within tissues. As demonstrated in the ESCC study, this approach to contextual imaging has the potential to uncover novel biomarkers and mechanisms involved in disease pathogenesis and treatment response. This should pave the way for more personalized therapeutic strategies.
Explore New Dimensions in Tissue Analysis with TissueGnostics’s Contextual Tissue Cytometry
At TissueGnostics, we understand the critical importance of precise and comprehensive tissue analysis in advancing medical research and improving clinical outcomes. Our StrataQuest Contextual image analysis platform is designed to transform your approach to tissue analysis through integrating detailed cellular and tissue-level insights with cutting-edge image analysis technology.
StrataQuest offers a solution for researchers seeking to gain a deeper understanding of the interactions within the tumor microenvironment. By utilizing our state-of-the-art software, you can unlock the full potential of multiplexed immunofluorescent staining data. Our tools can enable the precise quantification of cell density and detailed assessment of cellular distributions and interactions and much more.
Embrace the future of histological analysis by choosing StrataQuest for your research and diagnostic needs. Experience unparalleled accuracy and efficiency in tissue cytometry and propel your research into new frontiers. Additional information about image analysis, like immunofluorescence image analysis and image-based cytometry, is located on our blog. Our specialists are also available to help answer any questions, contact us for more details.