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 demands.
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.
This article discusses the main differences between classic cell counting with the famous hemocytometer and the automated Inish Analyser. You will learn the benefits of automatisation and when to choose one method or another depending on your lab needs.
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 are made at wide frequency ranges, providing information on cell size, membrane capacitance, and cytoplasm conductivity, along with cell concentration. The Inish Analyser is rated for cell concentrations between 50 thousand and 2 million cells per ml for SIM60.
Comparison between Inish Analyser and a Hemocytometer
First, a sample is prepared with an unknown high concentration, X cells per ml. The concentration is measured using a hemocytometer.
The Inish Analyser is rated for cell concentrations between 50,000 and 2 million cells per ml, so if your sample concentration is above 4 million cells per ml, dilute it down to 4 million cells per ml. This is labelled D0, and will be treated as the stock solution.
However, if your sample concentration is below 4 million cells per ml, just take that concentration as D0, and make sure that in the serial dilutions, your concentration never dips below the rated concentration of 50 thousand cells per ml.
With D0 prepared, the next step is serial dilution. D0 is diluted by half and labelled D1.
When preparing and measuring the concentration serial dilutions, the solution must be homogenous, so in order to avoid the error of having a sample separate into layers, the sample needs to be agitated using a vortex mixer, or by pipetting the solution up and down.
The solution is measured using a hemocytometer and using the Inish Analyser, three times. We are measuring the concentration at least 3 times to calculate the mean and standard deviation of the measurements for a rigorous comparison.
It’s important to take the 3 measurements in parallel; i.e you must measure the concentration first using a hemocytometer, then using the Inish Analyser, then using the hemocytometer, then the Inish again.
D1 is diluted by half again to make D2, which is again measured using a hemocytometer, and using the Inish Analyser at least three times as before.
This process is repeated for D3, D4 and D5.
Note: it’s important that the measurements are taken after every dilution, to avoid the error of time variability. The measurements won’t be clear if all of the serial dilutions are made, and measurements taken afterwards.
When you have finished taking all your data, in order to plot the values to see the correlation, you can find the mean and standard deviation for the measurement of each solution on both the hemocytometer and the Inish analyser, and plot them as shown.
Repeating this for every value and drawing a line of best fit should give you a straight line of y = x, showing that the Inish analyser is at least as accurate as the hemocytometry process. The smaller standard deviation for the Inish Analyser results shows that the Inish Analyser is much more precise, and doesn’t suffer from the same human errors found in using a hemocytometer.
The Inish Analyser gives results you can trust, empowering your research.
When choosing your cell counter, consider your lab's needs. Is it a large laboratory with future expansion plans? If this is the case, investing in an automatic cell counter can be highly beneficial.