Automated Cell Counter – Principle, Types, and Applications

An Automated Cell Counter is a machine or device that is used to count the number of cells in a biological sample. It uses various technologies such as image analysis, impedance measurement, or fluorescence to accurately determine the number of cells present.

The importance of cell counting lies in its widespread applications in the fields of medical research, cell biology, and clinical laboratory analysis. Knowing the number of cells in a sample is critical in determining the health and viability of cells, monitoring cell growth and proliferation, and in the development and testing of new treatments.

The history of cell counting dates back to the late 19th century when manual cell counting techniques using a microscope and a hemocytometer were developed. With advancements in technology and the increasing demand for more efficient and accurate cell counting methods, automated cell counters were developed in the late 20th century and have since become an essential tool in many biological and medical laboratories.

Cell counting is a standard laboratory technique for counting or quantifying cells, which is utilised in a variety of life sciences and medical diagnostic processes. There are numerous techniques to count cells with equipment, but they have been categorised as either manual or automated. For decades, the manual cell counter has been utilised. Nevertheless, its counting mistake and time-consuming limitations led to the development of more sophisticated equipment with automatic cell counting functions, known as automated cell counters.

A cell counter is laboratory equipment that counts cells automatically. The cells may have originated from humans, animals, plants, bacteria, viruses, or other sources. This device can offer an accurate cell count for a wider range of concentrations (from 5 x 10⁴/ml to 1 x 10⁷/ml).

Principle of Automated Cell Counter

Automated cell counters are essential tools in cell biology, biochemistry, and medical research. These instruments help in counting the number of cells in a given sample volume accurately and efficiently. In this article, we will be discussing the two distinct principles behind automated cell counters – the electrical impedance method and the optical method.

• Electrical Impedance Cell Counting: The Coulter Principle The electrical impedance cell counting principle, also known as the Coulter principle, is based on the increase in electrical resistance or impedance that occurs each time a cell passes through an aperture between two electrodes. The size of the aperture is the same as the cell size, and the change in impedance is directly proportional to the cell volume. This principle helps in counting the number of cells in a sample volume.

• Light Scattering Cell Counting: The Optical Method The optical method of cell counting, also known as the light-scattering cell counting principle, is based on the deflection of light due to cells. When a beam of light passes through a stream of diluted cells, it gets deflected, and the intensity and scatter angle of the deflected light depend on the cell type. A photometer detects the deflection, giving the number of cells present in a given sample volume.

• Sheath Flow Increases Detection Efficiency: In the light scattering principle of cell counting, sheath flow is applied, in which cells are hydrodynamically oriented. The application of sheath flow helps to increase the efficiency of detection.

In conclusion, automated cell counters have become indispensable tools in cell biology, biochemistry, and medical research. The electrical impedance method and the optical method are the two distinct principles behind automated cell counters, each with its unique advantages and disadvantages. Understanding these principles can help researchers choose the right instrument for their specific needs.

Types of Automated Cell Counters

On the basis of the aforementioned concepts, there are three types of automated cell counters;

1. Coulter counter

• These are particle counters based on the counting mechanism of electrical impedance cells.
• It provides the number of cells per particle and their size within the sample.
• They are unable to provide information regarding cell viability.
• It is useful for particle characterisation, haematology, and counting cells such as fat cells, plant cell aggregates, bacteria, etc.

2. Flow Cytometer

• Flow cytometer examines the characteristics of individual cells or particles in a stream by focusing light on one cell at a time. Therefore, distinct wavelengths of dispersed light and fluorescence signals are recorded.
• It is utilised for analysing cell shape, internal and exterior structure, and protein count.

3. Image-based cell counter

• It employs a bright-field or fluorescent microscope in conjunction with digital cameras, which are subsequently examined by image analysis software.
• For cell viability measurement, bright field-based cell counters employ colorimetric dyes, whereas fluorescent-based cell counters use fluorescent dye.

How to Operate an Automated Cell Counter

An automated cell counter is a sophisticated instrument that is used to count cells in a sample. However, these instruments work differently and thus it is essential to understand the operating procedure thoroughly. This article will provide you with a step-by-step guide on how to use an automated cell counter.

Preparation of Sample

Before using the automated cell counter, the sample must be prepared. Depending on whether trypan blue staining is used or not, the sample is prepared in two different ways:

• Without trypan blue staining: Pipette 10 µl of the sample directly.
• With trypan blue staining: Mix 10 µl of the sample with 10 µl of trypan and pipette 10 µl from the mixture for analysis.

Starting the Instrument

• Step 1: Turn on the instrument by pressing the power switch.
• Step 2: Handle the counting slide using the edges to avoid touching the optical surface of the slide.

Counting the Cells

• Step 3: Pipette 10 µl of the cell suspension into the outer opening of either chamber of the counting slide.
• Step 4: Insert the counting slide into a slide slot. The cell counter will automatically initiate the cell count.

Reading the Results

• Step 5: The count results will appear on the “Current Count” screen, showing the total cell count per ml. If the number of cells is above or below the specified range, the “Value Out of Range” message will be displayed on the screen.
• Step 6: To view the image, select the “View Image” key from the Current Count screen.
• Step 7: The dilution calculator calculates the volume adjustments needed to achieve the desired cell concentration.

Completing the Counting Procedure

• Step 8: Once the instrument completes the cell count, remove the slide.
• Step 9: Turn off the instrument after use.

An automated cell counter is a valuable tool for cell counting. Following the operating procedure correctly is crucial for accurate results. Ensure that you read the user manual that comes with the instrument carefully and familiarize yourself with the operating procedure before using the instrument.

Applications of Automated Cell Counter

• In Cell Culture Research: The viability of cell culture lines is a critical aspect of cell culture research. An automated cell counter can be used to determine the cell count and check the viability of the cell culture line being used.
• In Blood Analysis: Blood analysis is an important diagnostic tool used to determine the health condition of individuals. The concentration of various types of blood cells can be determined with the help of an automated cell counter. This information is critical in the diagnosis of various blood disorders and diseases.
• In Urine Analysis: Automated cell counters are also applicable in urine analysis. The number and types of cells present in a urine sample can be determined, providing valuable information about the individual’s health.
• In Cell Viability Measurement: One of the most important applications of automated cell counters is the measurement of cell viability. The fraction of viable and dead cells can be determined, providing critical information about the health of the cell culture.
• In Cell Therapy: In cell therapy, an automated cell counter is used to control the dose of cells administered to patients. This is critical in ensuring the safety and efficacy of the therapy.
• In Microorganism Growth Studies: Studies that examine the growth rate of microorganisms require cell counting. Automated cell counters are highly precise and can accurately count the number of cells, providing valuable information about the growth rate of the microorganisms.

In conclusion, automated cell counters have a wide range of applications in various fields. With their precision and versatility, they are a valuable tool in cell culture research, blood analysis, urine analysis, cell viability measurement, cell therapy, and microorganism growth studies.

Automated cell counter vs Hemocytometer

Feature	Automated Cell Counter	Hemocytometer
Accuracy	High	High
Speed	Fast	Slow
Cost	Expensive	Inexpensive
Ease of Use	Easy	Difficult
Sample Size	Small	Large
Versatility	Limited	Broad
Counting Method	Optical	Manual

An Automated Cell Counter is a machine that uses light scattering and imaging technology to count cells in a sample. They are fast, accurate and easy to use, but expensive and limited in versatility.

A Hemocytometer is a manual counting chamber used to count cells in a sample. It is less expensive, has broad versatility and can handle larger sample sizes, but is slower and requires manual counting, making it more difficult to use.

What is Countess ii fl automated cell counter?

The Countess II FL Automated Cell Counter is a cell counting instrument manufactured by Thermo Fisher Scientific. It uses fluorescence imaging technology to accurately count cells in a sample, making it well-suited for counting cells in complex suspensions and for use with a wide range of cell types. The Countess II FL Automated Cell Counter is fast, easy to use, and has a small sample volume requirement. It also has the capability to differentiate between live and dead cells, which is useful in certain cell biology applications.

Advantages of Automated Cell Counter

In the laboratory, accuracy and efficiency are critical factors. The manual cell counting method can be time-consuming and prone to human error, leading to inaccuracies in results. The automated cell counter is a great alternative for laboratories looking to automate their cell counting process. In this article, we will discuss some of the key advantages of using an automated cell counter in the laboratory.

• Accurate and Reliable Results in Less Time: One of the biggest advantages of using an automated cell counter is the accuracy and reliability of the results it provides. The automated process eliminates the risk of human error, providing results that are more accurate and reliable. Additionally, the process takes a fraction of the time compared to manual counting methods, making it a more efficient option.
• Reduces Workforce Requirements and Cell Count Variance: Automated cell counters significantly reduce the requirement of the workforce, making the process more efficient. The reduced workforce requirement means that the cell count variance is also significantly reduced, leading to more consistent results.
• Efficient and Cost-Effective: The automated cell counting process is more efficient and cost-effective compared to manual counting methods. The time savings and reduced workforce requirements result in significant cost savings, making it a more attractive option for laboratories.
• No Requirement for Replicate Counts at Low Cell Concentrations: Unlike manual counting methods, automated cell counters do not require many replicate counts at low cell concentrations. This results in further time and cost savings, making it a more attractive option for laboratories.

Limitations of Automated Cell Counter

Automated cell counters have revolutionized the way cell counting is performed in laboratories. However, despite its numerous advantages, there are some limitations to its use that should be considered. In this article, we will discuss some of the key limitations of using an automated cell counter in the laboratory.

• Results May Vary Due to Interfering Factors: One of the limitations of using an automated cell counter is that the results may vary due to interfering factors. For example, the presence of large particles, cell clumps, or bubbles in the sample can affect the accuracy of the results. Additionally, the presence of contaminants or other impurities can also impact the accuracy of the results.
• Expensive with High Running Costs: Another limitation of automated cell counters is the high cost of purchasing and maintaining the equipment. The cost of running an automated cell counter is also high due to the need for frequent calibration and maintenance. This can make it less attractive for some laboratories, especially those with limited budgets.
• Less Efficient in Detecting Atypical Cells: Automated cell counters can be less efficient in detecting atypical cells, such as toxic immature neutrophils. This can result in inaccurate results, leading to incorrect conclusions about the health of the cell culture or individual.
• Misclassification of Platelet Clumps: Automated cell counters can also misclassify platelet clumps as a single cell, leading to inaccurate results. This can result in platelets being misclassified as leukocytes or erythrocytes, leading to incorrect conclusions about the health of the individual.

Precautions

Automated cell counting is a crucial tool in various fields of research and diagnostics. However, to ensure the accuracy of results, some precautions must be taken while using the automated cell counter. This article will highlight some of the key steps you can take to ensure your cell counting results are accurate.

1. Proper Mixing of Cell Suspension: One of the most important steps in using an automated cell counter is to make sure that the cell suspension is mixed appropriately before pipetting. Failure to properly mix the cell suspension could result in inaccurate results. To avoid this, you can vortex the cell suspension or pipette it thoroughly to ensure the cells are evenly dispersed.
2. Safe Disposal of Slides: Automated cell counters use slides for cell counting, and it is important to dispose of these slides properly as they may contain biological material. Make sure to follow the safety regulations for disposal of biohazardous waste to avoid contamination and health risks.
3. Avoid Overfilling the Chamber: Another important step in using the automated cell counter is to avoid overfilling the chamber. Overfilling the chamber with sample could lead to biological contamination of the instrument, resulting in inaccurate results. To avoid this, make sure to use the correct volume of cell suspension as specified in the user manual.

By following these simple steps, you can ensure accurate results with your automated cell counter. This can help you save time and resources, while obtaining accurate and reliable data for your research or diagnostic needs.

FAQ

References

1. Green R, Wachsmann-Hogiu S. Development, history, and future of automated cell counters. Clin Lab Med. 2015 Mar;35(1):1-10. doi: 10.1016/j.cll.2014.11.003. Epub 2015 Jan 5. PMID: 25676368.
2. https://abu.edu.iq/sites/default/files/lbrary/20058.pdf