The Countleader FL 2000 is a fully automated, high-throughput cell counter that utilizes the patented FL24-plate with microfluidic channel design. It features automatic staining and supports AO/PI dual fluorescence staining mode, allowing analysis of 24 samples in just 17 minutes, or Trypan Blue staining mode in 12 minutes.

Principle  

The Countleader FL 2000 is an image-based detection system equipped with two fluorescence channels, integrating an advanced optical system and image recognition technology. It supports both classic brightfield (Trypan Blue staining) and fluorescence (Acridine Orange (AO) / Propidium Iodide (PI)) analysis modes, enabling simultaneous and accurate analysis of up to 24 samples. By combining statistical analysis with fluorescence imaging, it delivers precise information on cell concentration, cell viability, and cell morphology.

Automatic Staining Technology

FL24-Plate-A features a unique design with pre-embedded dyes. The AO/PI dyes are pre-loaded in the microfluidic channels. Simply add 40 µL of cell sample per well, and as the cells flow through the microfluidic channels, staining and analysis are performed automatically. This is ideal for high-precision counting of a large number of samples that need to be measured daily.

The FL24-plate-B is an empty 24-channel consumableplate. Only 20uL of cell sample and 20uL of TrypanBlue dye are needed per well. As the sample flowsthrough the microfluidic channels, automatic mixingand staining are completed simultaneously. lt is idealfor cell counting during the daily high-volume samplingstages of process development and optimization.

FL24-Plate Microfluidic Automatic Mixing and Staining System

Microfluidic Channel Preloaded with AO and PI Sample Loading Port Utilizing advanced microfluidic channel technology and an automated mixing design, this patented system effectively eliminates manual mixing errors, significantly improving sample counting efficiency and result accuracy!

Single-Use Design: Each sample remains isolated within the FL24-plate to prevent cross-contamination and ensure reliable experimental results.

No Cleaning or Maintenance Required: With no internal fluid pathways, the system requires no cleaning after use, greatly reducing operation time and maintenance costs.

Unique QR Code: Each FL24-plate is equipped with an individual QR code that automatically records and calibrates the analysis volume for each well, ensuring accuracy and consistency of results.

Simple Workflow, Automatic Analysis

Load the Sample

AO/PI Mode: Only 40 µL of cell sample per well is required. Compatible with multichannel pipettes, no pre-staining needed.

Trypan Blue Mode: Only 20 µL of cell sample and 20 µL of trypan blue dye per well, with no manual staining required.

Insert the Plate

Place the FL24-Plate into the designated slot of the Countleader FL 2000, select the wells to be counted, and start the analysis.

Automatic Staining and Analysis

As cells flow through the microfluidic channels, staining and analysis are performed automatically.

• Viable Cell Concentration
• Dead Cell Concentration
• Total Cell Concentration
• Cell Diameter
• Cell Aggregation Ratio
• Cell Fluorescence Images
• Cell Fluorescence Intensity Distribution Charts
• Cell Diameter Distribution Charts

Powerful and Flexible Software

• Supports both AO/PI and Trypan Blue analysis modes; capable of analyzing aggregated cells.

• All analysis data are automatically and continuously saved, with customizable programs and re-analysis functions based on cell status.

• 1.5× magnification, providing an analysis area 12 times larger than that of a traditional hemocytometer, resulting in higher sample volume and reduced statistical error.

• Data can be searched by experiment name, channel number, time, or sample ID.

• Individual sample IDs enable easy browsing and result searching.

• Automatic compensation for dilution factors.

• Generate cell growth curves to monitor and understand cell growth trends.

Supports proportional analysis across different cell size ranges, revealing patterns within heterogeneous populations — ideal for monitoring iPSC differentiation and virus-infected 293 cells.

The software is powerful, user-friendly, and highly flexible, allowing you to select detection channels based on the number of your samples.

It offers default analysis modes for both AO/PI and Trypan Blue staining and also allows users to create customized analysis methods tailored to their specific samples with one-click re-analysis of previously collected data.

Accurate Data with Excellent Reproducibility

Each sample is imaged at 1.5× magnification with automatic focusing, covering an analysis area of approximately 4.7 × 3.1 mm² — about 12 times larger than a traditional hemocytometer. This enables detec tion of tens of thousands of cells per sample. Combined with fully automated staining and analysis, it eliminates errors from manual operations.

CHO cell concentration gradient plot

This guarantees highly accurate and reliable results — ideal for counting CAR-T cells, PBMCs, low-purity cell populations, and irregular or compromised cells. No need to adjust parameters based on cell morphology or size, minimizing user-in duced errors.

PBMC Analysis in Brightfield and AO/PI Fluorescence Modes: Red blood cells and debris (indicated by red arrows in the left image) are not stained or counted in the AO/PI fluorescence images (middle and right images). Only cells containing nuclear DNA are stained and included in the counting and analysis.

Intuitive and Versatile Data Visualization 

Fluorescence images provide a clear, visual display of cell counting and identification.

The framed results indicate whether cells have been correctly recognized — AO (green) and PI (red).

AO/PI Dual Fluorescence Images

Easily generate growth curves from continuous culture data to monitor cell proliferation and refine your cell culture strategies.

Data Management  

Data Management

Obtain cell fluorescence images, flow cytome try–like scatter plots, and histograms.

Flexible Data Export Options

PDF, TIFF, Excel. 

USB 3.0 Output 

Supports export via USB drive, server, or printer. Equipped with at least one USB port and addition al USB expansion ports.

Large Built-in Storage 

2 TB internal storage, capable of saving over 25,000 data entries.

21 CFR Part 11 Compliance 

The Countleader FL 2000 features strict three-level user management and access control, fully compliant with 21 CFR Part 11 requirements.

• Password-protected user login

• User-specific access control

• Audit trail

• Log files

• Electronic records and signatures

IQ/OQ/PQ Services

The Countleader FL 2000 offers full IQ/OQ/PQ validation services, including a range of cell-simulating beads at various concentrations and comprehensive 3Q documentation. Designed to meet the rigorous demands of pharmaceutical manufac turing, and streamlines future regulatory submissions.

Countleader FL2000 offers three different types of consumables to meet various application needs

Applications

During cell line development and process development, there is a high volume of daily samples. Countleader FL2000 offers a high-throughput solution with low sample consumption, fast analy sis, and fully automated staining — dramatically improving counting efficiency!

• Process Development (PD) and Cell Line Development (CLD) in CGT, antibody, and vaccine research

• PBMC and CAR-T cell therapy

• Stem cells

• Tumor cells

• …