Cell studies have generally examined populations of cells in the thousands and millions. With the use of statistical models, this work has played a significant part in disease (i.e., cancer) research and discoveries. However, with the advancements of analytical techniques, many researchers have shifted their focus to single-cell analysis, which has created strong demand for single-cell isolation techniques.

Aside from manual pipetting of suspended cells, there are relatively few automated techniques for isolating single cells. Fluorescence activated cell sorting (FACS) is perhaps the most popular method due to its high-throughput capabilities. The disadvantage is that it requires large cell amounts and imposes stress on the cell that may compromise viability.

BD Biosciences offers the Influx cell sorter, which features up to 10 laser paths and 24 data acquisition channels. The system’s Computerized Cell Deposition Unit allows sorting into microtiter plates, slides or petri dishes, which is important for single-cell cloning, or sorting for single-cell PCR or single-cell imaging. Beckman Coulter offers the MoFlo XDP, which is equipped with the CyClone custom plate and slide sorting feature. Some researchers employ the LabCyte Echo acoustic liquid handling system after FACS to prepare libraries for whole genome and transcriptome analysis of genetic material from single cells.

Another method for single-cell sorting is laser microdissection (LMD) and optical tweezers. LMD systems isolate and collect cells of interest by combining a microscope, laser source (IR and/or UV) and software. It is low throughput but extremely useful for isolating a cell from fixed tissue. Key suppliers of LMD equipment are Carl Zeiss, Leica Microsystems (Danaher), Thermo Fisher Scientific and Molecular Machines.

Carl Zeiss offers the PALM CombiSystem, which combines LMD with optical tweezers. It employs a targeted laser pulse to catapult a cell into a collection vessel. Leica manufactures the LMD6 and LMD7, which uses a gravity-based collection method. Thermo sells the Applied Biosystems ArcturusXT, which utilizes a transfer film and two lasers. An NIR laser is used to melt the film onto the selected cells. With the cells attached to the film, the UV laser is used to cut out the specific cell(s) of interest. Molecular Machines offers several tools, including its MMI CellCut LMD, Cell Manipulator optical tweezers and the CellEctor capillary cell sorter.

One of the key applications for single-cell isolation techniques is NGS library preparation. Fluidigm’s C1 is a microfluidic-based single-cell isolating technology that can capture up to 800 cells, placing cells into individual reaction chambers. Cells can be stained and lysed for qPCR or sequencing.

The market for novel single-cell isolating techniques is burgeoning, particularly for single-cell genomic applications and NGS use. New systems that allow for increased automation, performance and efficiency are growing the market.

Single-Cell Isolation at a Glance:

Leading Suppliers

• Becton, Dickinson

• Beckman Coulter (Danaher)

• Carl Zeiss

Largest Markets

• Academia

• Biopharmaceuticals

• CROs

Instrument Cost

• $30,000–$400,000