Nexcelom Bioscience


Cellometer Auto X4 Cell Viability Counter

The X4 is a PC-based automated cell counter for 4x and 10x bright field and single channel fluorescent cell count, size, and viability analyses of mammalian cells, yeast, algae, and platelets. Advanced image analysis software provides one-step, size-based concentration and viability measurements using clumpy or irregularly shaped cells, excluding debris, with data in less than 60 seconds.

Automated Fluorescent Cell Counting in <60 Seconds

Cell Counting Made Simple

Automated cell counting made simple
fluorescent cell image analysis

The Cellometer Auto X4 utilizes bright field imaging, fluorescent imaging and pattern-recognition software to quickly and accurately identify and count individual cells. Cell count, concentration, diameter, and % viability are automatically calculated and reported.

5x Faster than Manual Counting 5x Faster than Manual Counting: Load sample, view image, count cells, and obtain results in < 60 seconds

The Cellometer Auto X4 Allows Hemacytometer Users to:

  • Increase throughput
  • Improve consistency
  • Ensure all data is correctly captured
  • Count difficult cells (clumpy, irregular-shaped)
  • Eliminate judgment errors, miscounts, and user-to-user variability

Fluorescent Analysis

Fluorescent dyes that stain DNA can be used to identify nucleated cells in a mixed cell population. Acridine orange (AO) is a nuclear staining (nucleic acid binding) dye permeable to both live and dead cells. It stains all nucleated cells to generate green fluorescence when imaged with the Cellometer Auto X4.

Because mature mammalian red blood cells don't contain nuclei, only mononuclear cells produce a fluorescent signal. Fluorescent-positive nucleated cells are easily counted. There is no need to lyse red blood cells, saving time and eliminating an extra sample preparation step.

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Trypan Blue Viability

Trypan Blue Viability

The Cellometer Auto X4 simultaneously calculates cell concentration and % viability for cultured cells stained with trypan blue.

Trypan blue viability is a dye exclusion method that utilizes membrane integrity to identify dead cells. The dye is unable to penetrate healthy cells, so they remain unstained. Dead cells have a compromised cell membrane that is permeable to the trypan blue dye. Dead cells are stained blue and display as dark cells in the Cellometer software with bright field imaging.

Concentration and Viability Results Within 30 seconds, the Cellometer Auto X4 instrument reports:
  • Live cell concentration
  • Percent viability
  • Mean live cell diameter
  • Live and dead cell number

Counted bright field Image

Counted bright field cell Image Live cells = green circle
Dead (stained) cells = red circle

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Fluorescent Viability Determination

Use of a fluorescent dye, such as propidium iodide (PI), is highly recommended for accurate viability analysis of cell samples containing debris.

Propidium iodide (PI) is a nuclear staining (nucleic acid binding) dye that enters dead cells with compromised membranes. It stains all dead nucleated cells to generate red fluorescence.

Because the Cellometer Auto X4 can count dead cells in the red fluorescence channel, there is no interference from debris and live cells.

With propidium iodide (PI), dead cells are clearly visible in the red fluorescence channel.

Fluorescent cell viability determination

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1-Step Yeast Concentration & Viability with Cellometer Auto X4 (10x)

Bright field and Fluorescent Imaging

1-step yeast cell concentration and viability
Bright field and fluorescent images of yeast cells. Cells appearing in both the bright field and fluorescent image are dead.

The Cellometer Auto X4 (10x) Fluorescent Cell Counter is specifically optimized for simple, 1-step determination of yeast concentration and viability. The Cellometer Auto X4 (10x) is ideal for research laboratories and small and large breweries looking to automate their fermentation monitoring. Performance of the Cellometer Auto X4 (10x) has been proven in the largest breweries in the U.S.

Bright field images can be viewed to confirm cell morphology. Counted bright field images show counted cells within cell clumps. Fluorescent images show dead cells for confirmation of viability results. Analysis of both bright field and fluorescent images from each sample makes 1-step concentration & viability determination possible. Read more about yeast viability.

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Analysis of Clumpy and Irregular-Shaped Cells

Including the NCI-60 human cancer cell lines developed by the National Cancer Institute.

Clumpy Cells

The MCF-7 breast cancer cell line can be very clumpy. The Cellometer pattern-recognition software identifies and counts individual cells within these cell clumps for accurate analysis (shown at right).

For clumpy cells with poorly-defined edges, the Cellometer Auto X4 offers fluorescent counting. Cells can be stained with a nuclear dye, such as acridine orange (AO), then imaged and counted in the fluorescent channel.

counted clumpy cell image
Count irregular-shaped cells

Irregular-shaped Cells

The Cellometer cell roundness setting can be adjusted for recognition and counting of irregular-shaped cells, such as RD cells and activated T-cells.

For clumpy cells with poorly-defined edges, the Cellometer Auto X4 offers fluorescent counting. Cells can be stained with a nuclear dye, such as acridine orange (AO), then imaged and counted in the fluorescent channel.

  • 57% of the NCI-60 cell lines are clumpy, contain debris, or display large variations in cell shape or size.
  • All 59 NCI-60 cell lines have been successfully validated on Cellometer Systems

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Algae Concentration using Cellometer Auto X4 (10x)

Bright feild algae cell image
Fluorescence algae cell imaging

Fluorescence imaging allows for simple, accurate concentration determination in algae samples containing debris.

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Exclusion of Debris & Size-Based Counting

Because Cellometer recognizes cells based on size, brightness, and morphology, cellular debris is easily and accurately excluded from counting results.

  • Cell size parameters can be modified to optimize exclusion of debris from results and enhance the accuracy of counting for a wide range of cell sizes.
  • A fluorescent nuclear dye, such as acridine orange, can be used to more easily identify nucleated cells in samples containing debris.
Counted cell image excludes debris
Cell size histogram based on cell diameter

The Cellometer Auto X4 Cell Counter automatically generates a cell size histogram based on cell diameter.

The minimum and maximum cell diameter settings can be optimized to count specific cells in a sample.

counted mature dendritic cells based on cell diameter
This example demonstrates counting of mature dendritic cells cultured from PBMCs based on cell diameter.

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View, Print, and Save Cell Images and Data Tables

counted cell images of mouse splenocytes
Bright field and fluorescent counted images of mouse splenocytes stained with acridine orange.

View bright field images to check cell morphology.

View fluorescent counted images to confirm exclusion of debris or unwanted cell types and correct counting of cells within clumps.

Export data to Excel

Easily Export to Excel for additional data formatting or sharing. Images and data tables can be pasted into PowerPoint presentations or submitted for publication.

Save data

Easily Save data and images to a network or local computer.

Print data

Print directly from the software screen. The standard printout displays live, dead, and total cell count and concentration, % viability, and cell images.

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User-Changeable Fluorescence Optics Modules

The Cellometer Auto X4 Cell Counter features a user-changeable fluorescence optics module enabling detection from UV to far-red. This enables the Cellometer Auto X4 to be shared by users performing a variety of assays for nucleated cell count, metabolic activity, and fluorescent viability.
User-Changeable Fluorescence Optics Modules
Easily switch optics modules for detection of a wide range of dyes and fluorophores:
375/450nm: DAPI, Hoechst
470/535nm: Acridine Orange (AO), Calcein, FITC
527/595nm: Ethidium Bromide (EB), Propidium Iodide (PI)
470/610nm: Chlorophyll
630/695nm: Allophycocyanin (APC)

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Counting Chambers: No Washing or Contamination

Cellometer Disposable Counting Chambers consist of two independent enclosed chambers with a precisely controlled height. Cell suspension of 20 microliters is loaded into the chamber using a standard single channel pipette.

The chamber is inserted into the Cellometer cell counter and the cells are imaged. This simple sample loading and analysis method is ideal for fragile cells.

Cellometer cell counting chamber
The disposable Cellometer Cell Counting Chambers offer several key advantages:
  • Time savings – no washing
  • No risk of cross-contamination
  • Reduced biohazard risk to users
  • Controlled sample volume
  • Most affordable automated counting consumables

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Dedicated On-line and On-site Applications Support

Nexcelom Technical Support Specialists

Experienced Nexcelom Technical Support Specialists are available from 9am to 5pm EST for phone and on-line support and can assist with:

  • Creation of new cell types
  • Optimization of counting parameters
  • Troubleshooting
  • Training of new users
  • Installation of a new Cellometer Auto X4 Cell Counter


The help button at the bottom right of the Cellometer Auto T4 software screen gives users instant access to:

  • Software features and instructions
  • On-line tutorials and training videos
  • Submission of a Support Ticket

Submit a Support Ticket

Clicking "Submit a Support Ticket" launches a convenient on-line form that is submitted directly to Nexcelom Technical Support

  • T4 instrument information fields are automatically populated
  • Images are easily attached for troubleshooting and applications support
  • A Nexcelom Specialist can assume remote control of the Cellometer Auto X4 Cell Counter for troubleshooting and training

Nexcelom Field-based Applications Specialists

Nexcelom Field-based Applications Specialists are also available for:

  • On-site demonstrations
  • Training
  • Troubleshooting
  • Technical Seminars
All Nexcelom Applications Specialists are 100% focused on image-based cell concentration & viability and cell-based assays using Cellometer Image-Based Cytometry

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Cellometer Auto X4 Accessories

Cellometer Auto X4 Fluorescence Optics Modules

Catalog # Description Compatible Dyes / Fluorophores*
XB-450-301 Cellometer Auto X4 Optical Module
Excitation / Emission: 375nm/450nm
DAPI, Hoechst 33342 and 33258
XB-535-401 Cellometer Auto X4 Optical Module
Excitation / Emission: 470nm/535nm
Acridine Orange (AO) (+DNA), Calcein, Fluorescein (FITC), SYTO®9, SYTO®13
XB-595-501 Cellometer Auto X4 Optical Module
Excitation / Emission: 527nm/595nm
Propidium Iodide (PI),Ethidium Bromide (EB), Rhodamine B, SYTOX® Orange
XB-610-401 Cellometer Auto X4 Optical Module
Excitation / Emission: 470nm/610nm
XB-695-601 Cellometer Auto X4 Optical Module
Excitation / Emission: 630nm/695nm
Allophycocyanin (APC)

IQ / OQ Validation Package

Installation Qualification (IQ) and Operational Qualification (OQ) package for the Cellometer Auto X4 Cell Counter

Catalog # Description Size Unit
Cellometer Auto X4-IQOQ Counting chambers, reference bead solution, and validation protocol for the Cellometer Auto X4 100 counts 1 Set

On-Site Service, Maintenance, and Instrument Warranties

Trained Nexcelom Applications Specialists are available for on-site preventative maintenance, IQ validation, and OQ validation.

Service Description
Preventative Maintenance Available for all Cellometer Systems
Please inquire for scheduling and pricing.
IQ Validation* Available for Auto T4, Auto X4, and Vision Systems
Please inquire for scheduling and pricing.
OQ Validation* Available for Auto T4, Auto X4, and Vision Systems
Please inquire for scheduling and pricing.
Extended Warranty Available for all Cellometer Systems currently under a valid warranty.
Please inquire for pricing.
*IQ/OQ Validation Package must be purchased separately

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