The value of quality samples
In cellular biology, downstream assay results are highly dependent on the quality of the preliminary samples used. Poor quality samples – such as those containing a high percentage of dead cells and debris – will produce poor results. It is the fundamental quality of the starting sample that ultimately dictates the quality of the results, and it is the sample preparation step which is often the most difficult to achieve successfully.
By utilizing the LeviCell® platform for cell isolation, scientists have consistently demonstrated robust enrichment for a wide range of target cells – without affecting cellular expression or original population representation.
Levitation Technology
LevitasBio® has developed an innovative Levitation Technology that runs a touch-free, label-free, three-step enrichment process on up to 96 samples at once with the LeviCell 96, or 12 with the LeviCell 12, with no centrifuge or beads. This can be applied seamlessly to a wide range of research areas and workflows.
Step 1: Input
Mix each sample with Levitation Agent, a paramagnetic solution, and pipette it into the wells of the SALT™ levitation array: up to 96 samples on the LeviCell 96 or 12 on the LeviCell 12.
Step 2: Enrichment
Place the array in a standard SBS deck position. In every well, viable cells levitate into a tight band above dead cells and debris during a 40-minute levitation, with no labels, beads or centrifuge.
Step 3: Collection
The liquid handler collects the enriched fraction from every well into an output plate, ready for counting and downstream assays, leaving dead cells, debris and supernatant behind.
*Illustrations show the LeviCell 96 System. The LeviCell 12 runs the same three-step workflow and chemistry for up to 12 samples per run.
Viable cells levitate much higher than dead cells and debris, allowing for clear separation and collection by the LeviCell systems. Start with high-quality cells every time for the best and most accurate results. This gentle yet effective process makes it possible.
- Maximize cell viability and enrichment
- Preserve cell population representation
- Minimize the risk of contamination and damage
- Maximize data for every sample analyzed
The path to high-quality results
The innovative label-free separation technology of the LeviCell systems facilitate complete debris and dead cell removal without affecting the original population representation of gene expression (please see our Population Representation application note for additional data on how the LeviCell maintains the original population representation).
When it comes to single-cell study, purity and viability of cells harvested from the sample preparation stage is a determining factor for generating high quality data in downstream assays such as NGS (Next Generation Sequencing).
Revolutionary viability and yields
LeviCell demonstrated the successful removal of dead cells and debris from a variety of primary samples while maintaining the original representation of the populations found within the sample. Furthermore, the LeviCell was able to enrich a low starting number of live cells to a final viability of >90%, exhibiting a capacity for scalability regardless of starting number of cells. 100,000 cells from each of two dissociated tumor cells (DTCs) were successfully processed and enriched to a final viability of >90% through removal of dead cells and debris in 3 steps and 20 minutes.
Superior performance over bead-based methods
The ovarian sample was split into two fractions of 100,000 cells and each fraction independently processed using either the LeviCell system or a popular bead-based dead cell removal kit. The output from each enrichment method was analyzed by manual cell counting using a Trypan Blue stain and a hemocytometer. LeviCell clearly outperforms the standard bead method in both Yield and Viability, while providing a faster, simpler and much more accommodating workflow when working with challenging samples. Additionally, LeviCell maintains the original cellular representation of the native sample without affecting the gene expression profile or activation state in even the most sensitive cell types.
Viability vs. Sort Method
Yield vs. Sort Method
No negative effects on post-processing population representation
The data below show how LeviCell’s gentle cell separation approach enables the study of notoriously delicate and sensitive primary cell types without compromising the integrity of the original cell populations. Sorted and unsorted ovarian DTC samples were blocked with TruStain human Fc Block for 15 minutes at RT then stained with 5 μL each of anti-CD45, anti-CD19, anti-CD11b, and 10 μL of anti-CD3, for 30 minutes at room temperature. Samples were washed with FACS buffer (0.5% BSA in PBS) once before analyzing on a Sony SH800S cytometer. The LeviCell output shows a similar amount of CD3+, CD19+, and CD11b+ cells within the CD45+ lymphocyte population while the sample processed by the bead method shows reduced proportions of CD3+ and CD11b+ cells.
No effect on the post-processing activation state
Macrophages, one of the most sensitive cell types, demonstrate the gentleness of the LeviCell system. The data below confirm there are no adverse side effects on cells’ post-processing activation state.
RAW264.7 cells were treated with IFN-g and IL-4 in order to polarize the macrophages into either the M1 or M2 stage. The samples were then processed on the LeviCell system. qRT-PCR was then performed to examine the expression of M1 specific gene (iNOS) or M2 specific (Arg1) in these cells to show that their activation states were not altered by the LeviCell. No differences in gene expression were observed between the input and output cells for the activation states.
Take your samples to the next level
The LeviCell systems’ ability to seamlessly enrich for target cells and produce robust yields of viable cells without preferentially depleting or changing the frequency or expression of cell types gives it the necessary technological characteristics that many scientists have been waiting for to take them to the next level.
Related Documentation
- Automated at scale
Viable cell enrichment, 96 wells at a time
The same label-free separation runs on the LeviCell® 96 System in a standard SLAS/SBS 96-well format. Load a plate, start a scripted protocol on the robot deck and walk away. Live cells levitate clear of dead cells and debris and are transferred to an output plate in the buffer of your choice, with no centrifuge, filters, beads or labels.
Enrich
Separate live cells from dead cells and debris in every well.
Wash
Exchange buffers in place, with no centrifugation steps.
Concentrate
Collect viable cells in a small volume, ready to load.
Plate
Deliver enriched samples to an output plate for any downstream assay.
96
samples enriched in 45 minutes
5 min
hands-on time
>95%
viability after enrichment, well after well
$10
per sample vs >$100 with standard methods
Every well is processed the same way, which removes operator-to-operator variability and makes large, consistent datasets practical, whether they feed a single study or the large models now being trained on millions of cells.