Rare Cell Isolation
Rare Cell Isolation - Related Videos
09:25
Detection and Enrichment of Rare Antigen-specific B Cells for Analysis of Phenotype and Function
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Cited by 36 •
2017
A simple yet effective method that employs magnetic nanoparticles to detect and enrich antigen-reactive B cells for functional and phenotypic analysis is described.
08:28
Targeted Metabolomics on Rare Primary Cells
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2024
Here, we present a protocol to accurately and reliably measure metabolites in rare cell types. Technical improvements, including a modified sheath fluid for cell sorting and the generation of relevant blank samples, enable a comprehensive quantification of metabolites with an input of only 5000 cells per sample.
09:49
Quantitative Measurement of Relative Retinoic Acid Levels in E8.5 Embryos and Neurosphere Cultures Using the F9 RARE-Lacz Cell-based Reporter Assay
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Cited by 6 •
2016
Methods to accurately measure retinoic acid (RA) levels in small amounts of tissue do not exist. This protocol describes the easy, quantitative measurement of relative RA levels in E8.5 embryos and neurospheres using an RA reporter cell line.
14:06
Detection of Rare Genomic Variants from Pooled Sequencing Using SPLINTER
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Cited by 10 •
2012
Pooled DNA sequencing is a fast and cost-effective strategy to detect rare variants associated with complex phenotypes in large cohorts. Here we describe the computational analysis of pooled, next-generation sequencing of 32 cancer-related genes using the SPLINTER software package. This method is scalable, and applicable to any phenotype of interest.
08:29
Profiling Epidermal Growth Factor Receptor DNA Mutations in Circulating Rare Cells Using a Cell Isolation System and Droplet Digital PCR
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2026
Here we present a protocol to illustrate the isolation of rare cancer cells from a contrived sample containing whole blood and spiked in lung cancer cells. The protocol utilizes a semi-automated, label-free cell isolation platform. Enriched cells are subsequently analyzed for epidermal growth factor receptor mutations using droplet digital PCR.
FAQs
They exploit measurable differences between the target and surrounding cells, including surface markers, size, density, deformability, and electrical behavior. Affinity-based capture and flow sorting emphasize cellular identity through surface features, whereas centrifugation and microfluidic separation can use physical properties. Selecting a discriminating feature determines which cells become enriched and which are removed.
Enrichment increases the proportion of the desired cells, while depletion removes unwanted populations that could interfere with analysis or culture. Using both strategies can improve target-cell recovery and reduce background contamination. This balance matters because concentrating cells is not sufficient if abundant non-target cells remain or if processing causes substantial target-cell loss.
Recovery and purity depend on how well the selected separation property distinguishes target cells from the rest of the sample. Surface-marker specificity, physical differences, and the chosen separation approach all influence the result. A method that strongly reduces contamination may also lose target cells, so researchers must consider both the amount recovered and the quality of the isolated population.
Affinity-based capture relies on surface markers, while flow sorting separates cells according to selected characteristics during controlled cell analysis. Centrifugation uses differences associated with density, and microfluidic systems can exploit properties such as size, deformability, or electrical behavior. These approaches provide complementary ways to isolate cells from complex biological samples.
A workflow generally begins with a complex biological sample, followed by one or more enrichment or depletion steps chosen for the target cell properties. The resulting fraction is then used for analysis or culture, with attention to contamination and cell loss. The isolated cells may subsequently support genomic, transcriptomic, or functional investigations.
Physical methods are useful when target cells differ measurably from background cells in size, density, deformability, or electrical behavior. Affinity-based capture is more appropriate when informative surface markers distinguish the population. The choice depends on which property is most reliably available in the sample and which approach best preserves the cells needed for later analysis or culture.
By concentrating scarce populations, the approach supports investigation of circulating tumor cells, stem cells, fetal cells, and defined immune cell subsets. Isolated cells can be examined through genomic and transcriptomic analyses or evaluated in functional studies and culture. This is especially valuable for characterizing heterogeneous populations and detecting biologically important cells that routine sampling may miss.