Empowering Single-Cell Research

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Concept

The QIAscout device together with its array is an affordable and accessible instrument enabling you to isolate and recover single cells.

  • Ideal for a variety of eukaryotic cells
  • Works in conjunction with inverted microscopes
  • Cells remain viable after separation
  • Isolated cells can be used for direct analysis or clonal expansion

Setup and Use

Setting up the QIAscout is easy and just takes minutes! Follow the guide in the video to see how easy it is to get your cells of interest.

Cell purity

  • The release needle and the magnetic wand do not act as sources of contamination while isolating single cells using the QIAscout. This was verified using two approaches, qPCR and Pyrosequencing, after processing cells using the REPLI-g Single Cell Kit.

    No DNA contamination observed with the release needle in qPCR analysis. A. qPCR amplification plots of 15 microrafts pierced using unwashed release needle. B. qPCR amplification plots of negative controls (water). C. qPCR amplification plots of positive controls (10 ng DNA) showing a CT value of 27.

  • The release needle and the magnetic wand do not act as sources of contamination while isolating single cells using the QIAscout. This was verified using two approaches, qPCR and Pyrosequencing, after processing cells using the REPLI-g Single Cell Kit.

    No cell contamination observed with the magnetic wand in Pyrosequencing analysis. A. Representative pyrogram showing the G>A point mutation in the human EGFR gene in a sample with single SW48 cells (positive control). B. Representative pyrogram showing absence of the A allele in single HT-29 cells, transferred with the same unwashed magnetic wand as the SW48 cells. C. Frequency of the A allele in two pierced HT-29 cells (negative control), two pierced SW48 cells (positive control) and 24 pierced HT-29 cells. Single SW48 cells transferred using a clean magnetic wand (positive control) show a mean frequency of A allele (=point mutation) of 50.7%. Pierced single HT-29 cells transferred using an unwashed magnetic wand and analyzed for the presence of SW48 cell contamination showed a similar frequency of A allele when compared to the pierced single HT-29 cells transferred using the clean magnetic wand (negative control), indicating no contamination between the two cell lines, the arrays and the microrafts.

Cell viability

  • Cells remain viable for several days and form subpopulations, irrespective of whether the pierced microrafts containing single cells are transferred to a cell culture dish or single cells present on microrafts are left intact and allowed to grow on the QIAscout array itself.

    Cells remain viable after piercing. A-B. Pierced microraft containing single SW48 cell. The microraft containing a colony of SW48 cells after several days of cultivation in a cell culture dish. C. Viable colony formed from a single MCF-7 cell. The position of the microraft in the cell culture dish is not important for further cultivation of pierced single cells.

  • Cells remain viable for several days and form subpopulations, irrespective of whether the pierced microrafts containing single cells are transferred to a cell culture dish or single cells present on microrafts are left intact and allowed to grow on the QIAscout array itself.

    Cells remain viable on the QIAscout array. Successful cultivation of a single HT-29 cell to form subcolonies on the QIAscout array after 90 hours confirms cell viability on the array.

Workflow data

  • Principle Component Analysis of miRNA expression of single cells. Differential response in miRNA expression amongst treated single cells with 3 distinct populations.

    Single-cell analysis for miRNA expression reveals heterogeneity across cell populations. In bulk cell analysis “cellular averages” mask intrinsic transcriptional variability. Distinct cell populations can be effectively revealed by single-cell analysis using the QIAscout for miRNA expression profiling.

Microscope compatibility

  • Measuring objective diameters to check for compatibility

    Determine the outermost diameter of the desired objective (ØD). The diameter should be between 18 and 30 mm to be compatible with the QIAscout release device.

    The minimum objective length (L) is the required length of an objective to ensure firm mounting of the release device to that objective.

  • The following microscopes and objectives have been tested and verified for fit and compatibility with the QIAscout instrument. This is not a complete list of all objectives. If the objective needed is not listed here, please refer to the next table.

  • Objective dimensions compatible with QIAscout release device

    The following table shows which insert to use based on the diameter of your microscope objective.

ZEISS® Primovert

Objective Cat. No. Magnification Insert release device Stage* Holding frame
Plan-Achromat 4x/0.10 for Primo 415500-1600-001 4x 4 (3)‡‡ Microscope specific stage compatible. It is recommended to install an object guide Universal mounting frame
Plan-Achromat 4x/0.10 Ph0 for Primo 415500-1619-000 4x 4 (3)

*Check with microscope manufacturer for appropriate stage. Check with microscope manufacturer for suitable holding/mounting frames and object guides. ‡‡ Either of the 2 sizes of the clamp inserts is usable. Note: w.o. stands for without.

ZEISS® Axio Vert.A1/Axio Observer

Objective Cat. No. Magnification Insert release device Stage* Holding frame
A-Plan 5x/0.12 M27 421030-9900-000 5x w.o. Change of stage might be necessary due to required stage opening size Universal mounting frame
A-Plan 5x/0.12 Ph0 M27 421031-9910-000 5x w.o.
A-Plan 5x/0.12 Pol M27 421033-9900-000 5x w.o.
EC Plan-Neofluar 5x/0.16 M27 420330-9901-000 5x w.o.
EC Plan-Neofluar 5x/0.16 Ph1 M27 420331-9911-000 5x w.o.
EC Plan-Neofluar 5x/0.16 Pol M27 420333-9901-000 5x w.o.
EC Epiplan 10x/0.25 M27 422040-9902-000 10x w.o.
Objective LD A-Plan 5x/0.15 Ph1 M27 421231-9910-000 5x w.o.
EC Epiplan 5 x0.13 M27 422030-9901-000 5x w.o.
Epiplan 10x/0.23 W0.8" 442030-9903-000 10x 3

*Check with microscope manufacturer for appropriate stage. Check with microscope manufacturer for suitable holding/mounting frames and object guides. ‡‡ Either of the 2 sizes of the clamp inserts is usable. Note: w.o. stands for without.

Leica® DMi1, DMIL LED or DMi8

Objective Cat. No. Magnification Insert release device Stage* Holding frame
N PLAN 5x/0.12 PH0 11506303 5x 1 DMi1 and DMIL LED: microscope specific stage compatible. It is recommended to install an object guide for fixed stages

DMi8: Change of stage might be necessary
Universal holding frame
N PLAN 5x/0.12 11506302 5x 1
HI PLAN 10x/0.25 11506228 10x 2
HI PLAN 10x/0.25 PH1 11506230 10x 2
HI PLAN 10x/0.25 POL 11556061 10x 2
HI PLAN 10x/0.25 SL 11506229 10x 2
HI PLAN EPI 10x/0.25 11566069 10x 2
HC PL Fluotar 5X/0.15 11506504 5x w.o.
HC PL Fluotar 10X/0.30 11506505 10x w.o.
HC PL FLUOTAR 10x/0.32 11506521 10x w.o.

*Check with microscope manufacturer for appropriate stage. Check with microscope manufacturer for suitable holding/mounting frames and object guides. ‡‡ Either of the 2 sizes of the clamp inserts is usable. Note: w.o. stands for without.

Nikon® EclipseTS2/Ti-U/Ti-S

Objective Cat. No. Magnification Insert release device Stage* Holding frame
Plan Fluor, 4x MRH 20041 4x w.o. Additional rectangular mechanical stage stroke might be necessary Terasaki Holder (for 65 mm dish)
Plan Fluor, 10x MRH 20101 10x w.o.

*Check with microscope manufacturer for appropriate stage. Check with microscope manufacturer for suitable holding/mounting frames and object guides. ‡‡ Either of the 2 sizes of the clamp inserts is usable. Note: w.o. stands for without.

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What our customers say…

"QIAscout turned out to be a very suitable system allowing us to select and transfer our cell of interest with nearly 100% efficiency and survival rate for further downstream application."Dr. Lajos Kemeny, University of Szeged

"With the QIAscout, researchers will be able to control exactly which cell to spend analysis resources on, …"Christoph Ziegenhain, Ludwig-Maximilians University, Munich

"…overcoming bulk averaging of cell populations and learning more about the different cell types present in biological samples."Christoph Ziegenhain, Ludwig-Maximilians University, Munich