NOBEL-SPA – NanOstirBar EnabLed Single Particle Analysis by confocal fluorescence microscopy

Cells communicate with each other through tiny structures called extracellular vesicles (EVs). These microscopic vesicles, each measuring less than 200 nanometers in size, play crucial roles in transmitting signals between cells. However, due to their heterogeneity and limited cargo capacity, studying EVs has posed significant challenges for researchers. In a recent breakthrough, researchers at the University of California-Riverside have developed a novel technique called NanOstirBar (NOB)–EnabLed Single Particle Analysis (NOBEL-SPA), promising rapid and accurate inspection of single EVs and shedding new light on their role in health and disease.

Unlocking the Potential of NOBEL-SPA

The NOBEL-SPA technique described in the present study revolutionizes the way we study EVs by enabling high-confidence examination of individual vesicles using confocal fluorescence microscopy. This innovative approach allows researchers to assess the colocalization of specific protein and microRNA (miRNA) markers within EVs derived from various cell lines or clinical serum samples.

NOB to facilitate single vesicle analysis

(A) Scheme of NOBEL-SPA. (B) Plot of EV numbers remained in the supernatant after incubation with NOBExo for various durations as detected by NTA. Value and error bars are the average and SD of three repeated measurements. (C) WB results for detection of CD63, CD81, and CD9 in the starting solution and in the EVs pulled down by NOBExo or by MBs with diameter around 1 (MB1) or 0.22 μm (MB2) which were conjugated with the same antibody mixture as NOBExo, after 30-min incubation. The first lane was from a protein ladder. (D) SEM images of the individual NOB, NOBExo, and NOBExo with the bound EV (pointed by the arrows). Scale bars, 100 nm. (E) Histograms for the pixel numbers of the fluorescent particles detected after EV lysis and cargo crosslinking, with the EVs captured on the glass slide surface (without NOB) or by NOBExo (with NOB). The inset images: 10 μm by 10 μm. Scale bars, 1 μm.

Discovering EV Subpopulations

Through NOBEL-SPA, researchers have identified distinct subpopulations of EVs characterized by the colocalization of unique protein and miRNA combinations. Remarkably, these EV subpopulations have shown the potential to detect early-stage breast cancer (BC), providing a promising avenue for early diagnosis and intervention.

Expanding Possibilities

The versatility of NOBEL-SPA extends beyond the study of EVs in breast cancer. This groundbreaking technique can be adapted to analyze other types of cargo molecules or small submicron biological particles, opening new avenues for research in diverse fields of biology and medicine. By studying how specific cargos are sorted into heterogeneous vesicles under different physiological conditions, researchers can gain valuable insights into EV biogenesis and identify novel therapeutic targets.

Implications for Clinical Practice

The development of NOBEL-SPA holds immense promise for clinical applications, offering a non-invasive and highly sensitive method for diagnosing diseases such as breast cancer at an early stage. Furthermore, the ability to analyze EV subpopulations may lead to the development of targeted therapies tailored to individual patients, revolutionizing the field of personalized medicine.

In conclusion, the NOBEL-SPA technique represents a significant advancement in our understanding of extracellular vesicles and their role in cell communication. By providing a powerful tool for the analysis of EVs, NOBEL-SPA has the potential to drive groundbreaking discoveries in biology and medicine, ultimately improving outcomes for patients worldwide.

Li Z, Guo K, Gao Z, Chen J, Ye Z, Cao M, Wang SE, Yin Y, Zhong W. (2024) Colocalization of protein and microRNA markers reveals unique extracellular vesicle subpopulations for early cancer detection. Sci Adv 10(9):eadh8689. [article]

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