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Hepatocyte

Primary Hepatocyte Viability & Live/Dead Cell Concentrations

  • Fast, Accurate Hepatocyte Count & Viability

  • Importance of Accurate Hepatocyte Counts

  • Limitations of Manual Cell Count & Solution

  • Cell Count & Viability by Dual Fluorescence

  • What do I Need to Get Started?

Fast, Accurate Primary Hepatocyte Count & Viability

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Cellometer provides cell sample analysis results that include:

  • Cell count
  • Cell concentration
  • Cell viability
  • Cell diameter and size distribution
  • Cell images, and more

Benefits of Cellometer Dual Fluorescence Viability Assays:

  • Only 20 µl of sample required, important for rare samples
  • Improve accuracy and eliminate errors
  • Ensure user-to-user consistency
  • Store data automatically

Importance of Accurate Primary Hepatocyte Counts

Primary hepatocytes are regularly used for determining the toxicity of drug candidates during the drug screening process. Cytoxicity assays are plate-based assay requiring accurate cell concentration and viability measurements so that the assays can be set-up correctly. Since primary hepatocytes are generally less than 80% viable and tend to form clusters, precise and accurate cell concentration and viability measurements are crucial to obtaining reproducible, high quality data.

Characteristics of Cytoxicity experiments involving primary hepatocytes
  • Hepatocytes lose viability over time, even with special care
  • Multi-well plates are used to screen drug candidates at different concentrations with repeated measurements, representing a large number of data points for analysis
  • Multi-well plates need to be seeded with identical numbers of cells with known viability for correct interpretation of data

Limitations of Manual Counting & Cellometer Solution

Challenges

Reliable concentration and viability data of primary Hepatocytes is critical for accurate analysis of compound toxicity in vitro. Due to hepatocytes' variable morphology, fragile nature and tendency to clump, traditional manual counting methods are time consuming and the subjectivity from operator-to-operator can cause inconsistent results.

Solution

Nexcelom's new method incorporates staining primary hepatocytes with a ready-to-use fluorescent dual staining solution that stains live cells with acridine orange, and dead cells with propidium iodide and then loading 20µL of labeled sample into a disposable counting chamber for analysis. Since the counting chamber is disposable, no washing is required between samples, and the risk of cross contamination is eliminated.

Fluorescent images of the stained cells are captured and using proprietary algorithms, Cellometer Vision's robust operating software accurately analyzes cell images to generate live cell count, concentration and viability percentage. Total analysis time typically takes less than 60 seconds. Cell images and all analysis data, including cell size distribution histograms, can be instantly saved for documentation. Data can also be easily exported to Microsoft Excel spreadsheets for further analysis.

Live and Dead Cell Image Primary Hepatocyte Cellometer Vision is capable of analyzing many species of primary hepatocyte including:
  • Human
  • Mouse
  • Horse
  • Rat
  • Rainbow Trout
  • Monkey
  • Dog
  • Rabbit
By overlaying AO/live and PI/dead hepatocyte images, counting results can be visually confirmed. Live hepatocytes are circled in green and dead hepatocytes circled in red.

Cell Count & Viability by Dual Fluorescence

Dual Fluorescence Viability Assay Principle

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How Cellometer Vision Does Dual Fluorescence Measurement

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What do I Need to Get Started?

Vision and related products

Request a Demo Request a Demo One of our Applications Specialists can provide a 20-30 miniute online demo or arrange a live demo in your lab.

Request a Demo Ask a Specialist Our Applications Specialists are available to answer your questions.

Cellometer Vision System for Primary Hepatocytes
Instrument
Cellometer Vision, 5x Magnification
Optics Modules
VA-535-401
VA-660-501
Counting Chambers
CHT4-SD100
CHT4-PD100
Reagent Kit
Cellometer ViaStain™ AOPI Staining Solution in PBS