This article discusses the technique of single-cell sampling for isolating live cancer cells from a suspension for further analysis. The process involves using a micromanipulator and microcapillary tip to collect individual cells with precision.
Single-cell sampling enables isolating a single live cell from a suspension of cancer cells for downstream analysis.
Begin the isolation with a cancer cell suspension. Transfer the desired volume of this suspension onto a glass slide. Leave the slide undisturbed for a while to reduce particle movement and allow the cells to settle.
Now, transfer the slide under a microscope equipped with a micromanipulator connected to a microcapillary tip. This high-precision instrument facilitates physical interaction with cells in the sample at a microscopic level.
Direct the microcapillary tip close to a single target cell. By applying suction, collect the target cell into the microcapillary. Now, transfer the microcapillary directly into a tube containing the desired collection buffer.
Eject the microcapillary contents to release the captured single cell into the buffer. Store the isolated live single cell on ice until further analysis.
For micromanipulation, transfer the entire 100 microliters of cell suspension onto a glass slide. Then, place the glass slide on a microscope equipped with a micromanipulator and allow the cells to settle for 5 minutes. Then, in 0.2-milliliter PCR tubes, pipet 2 microliters of cell lysis master mix and transfer on ice. Use the micromanipulator to collect a single cell in 1 microliter of 1X phosphate-buffered saline.
Then, transfer the cell directly into 2 microliters of cell lysis master mix. Use a desktop microfuge to quickly spin down the samples at 2000 x g for 3 seconds to collect the liquid at the bottom of the tube.
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