How to analyse data using the Inish Analyser?

Introduction

If you work with cells, you probably spend hours in the lab counting them.

As much as researchers love bench work, it can be handy to automate certain processes, especially when it comes to larger laboratories with high throughputs.

To solve this problem Cellix has recently released the Inish Analyser. It is a simple, automated and easy to use instrument for cell counting and viability. In this article, we will look at the data analysis protocol for the Inish Analyser. We will show you how to zoom in and out of your data on the Inish Analyser, and how to gate populations, in other words, adjust the cut-off points for the different populations.

Automated Cell Counting

As the name says, automated cell counters like the Inish Analyser are devices that can count cells automatically. It is a pretty straightforward process - you load your sample into the cell counter, which aspirates or pumps your cell sample through a small tube. These devices typically use optical or electrical impedance sensors or image-based analysis to count how many cells go through the tube.

The main features and benefits of the Inish Analyser include:

Label-free: No fluorescent stains or dyes.
Single-cell Analysis: Every cell is counted!
High-throughput: Analyse thousands of cells per second.
Compatible with a wide variety of cell samples: cell lines (e.g. Jurkats, CHO cells etc.), primary cells (e.g. PBMCs), stem cells, yeast cells, sperm cells.
Fast Set-up & Analysis: 90 seconds including sample preparation.
Easy-to-use Bench top Instrument: fits inside standard biosafety cabinet and operated by simple touchscreen workflow.
Compatible with FCS & CSV file format: easily exportable.

This method offers reliable cell counting with high precision and throughput.

Cellix's Inish Analyser uses impedance spectroscopy, a method based on differences in the electrical impedance between the cell and a buffer. It has a distinct advantage over other aforementioned methods – it’s label free! What does that mean? It means no fluorescent dyes or labels to stain or tag the cells for detection.

How does it work?

Cells flow through a microfluidic channel with embedded electrodes which form a current path. As the cells flow past the electrodes, they disturb the electric field resulting in a change in impedance. In this method, impedance measurements can be made at wide range of frequencies, providing information on cell size, membrane capacitance, and cytoplasm conductivity, along with cell concentration. The Inish Analyser is rated for cell concentrations between 50,000 - 2,000,000 cells/mL.

Data Analysis Protocol for Inish Analyser

Zoom in and zoom out

After running an assay, a graph and table is displayed showing the default results.
To zoom in or out, click anywhere on the amplitude or phase axes, and a window will pop out.
You can adjust the bounds of the axes by dragging the sliders or entering in values in the boxes provided.
Once you’re happy, press ‘okay’, and the changes will be applied.

Gating Populations

Now we’ll look at gating for noise or debris population:

To adjust the gate for noise or debris population, select ‘Noise’ from the dropdown menu in the upper-right corner of the graph.
The black line will be highlighted. You can drag this line to adjust the cutoff point for what is considered noise in your data.
Noise is generally represented as a straight vertical line in your data.
When you’re happy with the adjustment, select ‘Apply’ from the dropdown menu.
The population to the left of the line will turn black, and will be excluded from the concentration value displayed on the right.

Now we’ll look at gating for Live and Dead populations :

To adjust the gate for Live and Dead populations, choose Live/Dead from the dropdown menu in the upper right corner of the graph.
This will highlight the red line. You can drag this line to adjust the cutoff point for what are considered Live or Dead.
This is useful for when you want to consider strictly live or strictly dead, using a ‘worst-case-scenario’ approach.
When you’re happy with the adjustment, select ‘Apply’ from the dropdown menu.
The population above the line will turn green to indicate the Live population, and the population below the line will turn red to indicate the Dead population.
This will update the viability value displayed on the right.

The Inish Analyser gives results you can trust, empowering your research.

When choosing your cell analyser, consider your lab's needs. Is it a large laboratory with future expansion plans? If this is the case, investing in an automatic cell analyser can be highly beneficial.

If you'd like to learn more about Cellix's Inish Analyser, contact Cellix to book an online demo or request a quote.