Fine Needle Aspirate

New Standard for FNA Sample Processing

Introduction

The collection of guided fine needle aspirates (FNA) from tumors is a well-established clinical procedure used to help identify various lymphomas. The integrity of the process is influenced by several factors, including the delicate practice of sample processing for downstream analysis. Since there are no universal guidelines for handling FNA samples, diagnostic accuracy and test sensitivity vary widely among institutions.

Consequently, low cell count, diminished viability, high levels of debris, and population representation stability are constant challenges faced by scientists.

By consistently producing highly viable and replicable results from limited FNA samples, the LeviCellTM provides a cost-effective and time-saving solution that can potentially set the standard for isolating viable cells for FNA sample processing.

The LeviCellTM Solution

As the world’s first label-free cell separation technology, the LeviCell leverages the size and density of cells to isolate them from other cell types.  The LeviCellTM gently displaces target cells using less than 0.5 psi of force, which has proven to have no negative effects on even the most sensitive cell types. In comparison, conventional methods exert 70 – 100 psi of force, which damages hearty cells and completely destroys sensitive ones. The LeviCellTM is the only label-free technology in the market that can isolate and enrich a robust quantity of viable cells from high-value primary samples and sensitive cell types without affecting original population representation.

Using the LeviCellTM involves three easy manual steps. First, pipette the FNA sample directly from the collection tube into the single-use, biocompatible, sterile consumable.  Second, load the cartridge into the LeviCellTM platform. The LeviCellTM automatically processes the sample and separates live cells from dead, which will levitate at a lower height and is collected in the bottom channel of the consumable cartridge.  Live cells will be collected in the top channel.  This process will take approximately 30 minutes to complete.  Upon completion, collect target cells for downstream analysis, which is the third step of the LeviCellTM cell separation workflow.  Simple. Quick. Easy.

A five-fold increase in Viability

FNA samples were collected from human H358 tumor xenografts in rats by Hera Biolabs. An FNA sample was suspended in Levitation BufferTM and enriched for live cells using the LeviCellTM. A portion of the sample was set aside. The live channel output and the unsorted sample were stained with Trypan and counted on a microscope. The LeviCellTM output shows a nearly 5-fold increase in viability compared to the input.

Debris-Free Sorting

FNA samples from before and after sorting on the LeviCell were stained with Trypan and counted on a microscope. The images show a reduction in the number of dead cells and debris after sorting with the LeviCell.

No negative effects on population representation

To demonstrate the LeviCellTM’s gentle process – which enables robust yields without affecting population representation of the original sample – PBMCs were suspended in Levitation AgentTM and enriched for live cells using the LeviCellTM. A portion of the original sample was analyzed independently. Both live and dead channel outputs as well as the unsorted sample were blocked with TruStain human Fc Block for 15 minutes at RT then stained with 5 μLeach of anti-CD45 (PE), anti-CD3 (FITC), anti-CD11b (APC), and 10 μL of anti-CD19 (PerCP-Cy5.5) for 1 hour on ice. Samples were then washed with a FACS buffer (0.5% BSA in PBS) once prior to analysis on a Sony SH800S cytometer.

The live channel output (red bar) shows a similar amount of CD3+, CD19+, and CD11b+ cells within the CD45+ lymphocyte population as was present prior to sorting (blue bar). The consistency of robust live cell yield across all cell types is testament to how the LeviCellTM platform’s fast, gentle, and powerful separation technology enables the preservation of original population representation.

Conclusion

The LeviCellTM is uniquely positioned to transform the FNA sample processing workflow by delivering consistently robust and replicable results from extremely sensitive and limited FNA samples, without negatively affecting original population representation.  The revolutionary cell separation technology negates the need for markers, manual handling, and exertion of force greater than 1 psi – any of which can negatively affect cells and influence downstream results.  This is a significant development in the field, as FNA samples are difficult to procure, challenging to work with, and hard to replace. The LeviCellTM not only evens out the playing field when it comes to FNA sample processing, it completely changes the game by setting the standard.