Single Cell Electrophoresis
Single Cell Electrophoresis - Related Videos
05:06
Alkaline Single Cell Gel Electrophoresis: A Sensitive Technique for Quantitative Estimation of DNA Damage in Individual Cancer Cells Following Chemotherapy
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2025
In this video, we demonstrate an alkaline single-cell electrophoresis assay to quantify both single- and double-strand DNA breaks in doxorubicin-treated cells. Alkaline conditions disrupt the hydrogen bonds between DNA base pairs, resulting in the complete unwinding and separation of DNA strands. This allows differential migration of undamaged and damaged DNA fragments in agarose gel upon electrophoresis, revealing a comet-shaped structure of slowly migrating undamaged DNA forming the circular...
07:31
Two-Dimensional Gel Electrophoresis
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2023
Two-dimensional gel electrophoresis (2DGE) is a technique that can resolve thousands of biomolecules from a mixture. This technique involves two distinct separation methods that have been coupled together: isoelectric focusing (IEF) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). This physically separates compounds across two axes of a gel by their isoelectric points (an electrochemical property) and their molecular weights. The procedure in this video covers the main...
08:50
Capillary Electrophoresis (CE)
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2023
Source: Laboratory of Dr. B. Jill Venton - University of Virginia Capillary electrophoresis (CE) is a separation technique that separates molecules in an electric field according to size and charge. CE is performed in a small glass tube called a capillary that is filled with an electrolyte solution. Analytes are separated due to differences in electrophoretic mobility, which varies with charge, solvent viscosity, and size. Traditional electrophoresis in gels is limited in the amount of voltage...
09:22
DNA Gel Electrophoresis
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2023
DNA gel electrophoresis is a technique used for the detection and separation of DNA molecules. An electric field is applied to a gel matrix comprised of agarose, and within the gel, charge particles will migrate and separate based on size. The negatively charged phosphates of the DNA backbone cause DNA fragments to move toward the anode - a positively charged electrode. The video explains the mechanism by which DNA fragments are resolved on an agarose gel, and it provides a step-by-step...
04:43
Single-Cell Western Blotting to Determine Cell Heterogeneity
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2025
The video describes a single-cell western blotting technique that identifies the composition of individual cells captured on a single-cell western blotting chip in a heterogeneous suspension. The technique detects the fluorescence intensity of the fluorophore tagged to a protein marker specific to the target cell.
FAQs
Alkaline and neutral conditions provide different experimental settings for examining DNA damage, while the assay's basic readout remains migration away from the nucleoid. Because the overview identifies both formats without assigning them to particular lesions, researchers should interpret tail measurements in relation to the selected condition. This distinction helps compare experiments without treating all comet patterns as interchangeable.
Tail length and tail intensity are complementary readouts rather than interchangeable labels. In the comet pattern, DNA that has moved away from the nucleoid produces the tail, and the dimensions and strength of that signal reflect strand breaks. Using both features gives researchers a more informative description of damage in each cell than relying on a single visual characteristic alone.
Single-cell measurements reveal whether DNA damage is distributed broadly or concentrated in a subset of cells. That distinction matters because a bulk result can average together cells with very different damage levels, obscuring genetic heterogeneity. In cancer research, examining individual comet patterns can therefore support more precise comparisons between tumor-cell populations and help identify variation that collective measurements might hide.
The core workflow preserves cells in an agarose matrix, removes cellular material through lysis, and then applies an electric field under a chosen alkaline or neutral condition. Damaged or fragmented DNA migrates from the nucleoid during electrophoresis, producing the comet-like pattern that is measured. Keeping these stages conceptually distinct helps relate the final signal to preparation and field exposure.
When the goal is to study genotoxic exposure, the assay can compare DNA damage at the level of individual cells rather than only across a pooled sample. This makes it useful for evaluating how an exposure affects tumor-cell populations and for examining whether damage differs among cells. The resulting single-cell profiles add context to cancer studies of tumor biology.
Treatment studies can use the assay to examine DNA damage after chemotherapy or radiation and to compare DNA repair capacity among tumor cells. Those measurements connect a cell-level damage pattern with questions about treatment efficacy and resistance. The method is therefore relevant not only to whether therapy produces damage, but also to how differently tumor cells handle that damage.