Imaging Flow Cytometry (IFC)

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Imaging Flow Cytometry (IFC)

Method Introduction

Imaging flow cytometry (IFC), sometimes called multispectral imaging flow cytometry (MIFC), combines two techniques: flow cytometry and imaging microscopy.

Like a regular flow cytometer, IFC obtains forward and side-scatter information and records several fluorescence signals simultaneously from micrometer-sized particulate objects in a sample stream. In IFC, however, these data are recorded using a high-resolution microscope set-up, allowing the subsequent visualization of brightfield, darkfield, and fluorescence patterns on individual or overlaid images.

As a rather novel technique in particle characterization, IFC is mainly used as a research tool supporting more established techniques, such as flow imaging microscopy, during particle identification and troubleshooting. A clear benefit over other orthogonal techniques is the potential to connect particle morphology and fluorescence information on a particle-by-particle basis.

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Applications

Imaging flow cytometry is typically used to analyze whole cell populations but can also be employed to analyze protein particles or other particulate matter present in biopharmaceutical drug products.

Quality and Biosafety Level

We provide all our analytical services with the highest quality standards. Experienced scientists carry out each project, and a scientific reviewer comprehensively checks every report or data presentation.

We offer this technology with the following quality and biosafety levels:

R&D level

We offer this method under R&D. Our GRP system assures the highest-quality research standards.

Up to biosafety level 1

This method can be applied to proteins, nucleic acids, and most viral vectors, including AAVs and more.

Imaging Flow Cytometry (IFC) Frequently Asked Questions (FAQs)

  • Imaging flow cytometry (IFC), also known as multispectral imaging flow cytometry (MIFC), is a hybrid analytical technique that combines the quantitative power of flow cytometry with high-resolution image capture. It enables simultaneous acquisition of brightfield, darkfield, and fluorescence images to support detailed particle analysis.

  • IFC supports particle identification and morphological assessment in biologic drug products by visualizing individual particles and measuring their size, shape, and fluorescence. It is a valuable tool for troubleshooting, comparability assessments, and formulation optimization.

  • Unlike traditional flow cytometry, IFC captures actual images of each particle or cell analyzed. This enables both qualitative and quantitative evaluations, improving resolution and allowing for direct visualization of structural features and fluorescent signals.

  • Coriolis uses IFC to analyze protein particles, viral vectors (such as AAVs), nucleic acids delivery systems, and other subvisible particles within complex biologic formulations. The method is suitable for BSL-1 materials.

  • At Coriolis, IFC is currently offered under non-GMP (R&D) conditions within a Good Research Practice (GRP) environment. All studies are executed by trained scientists with rigorous data review procedures in place.

  • Yes, IFC is designed to detect and characterize micrometer-sized particles, making it well suited for subvisible particle analysis. It complements techniques such as micro-flow imaging (MFI), light obscuration (LO), and dynamic light scattering (DLS).

  • Fluorescence channels allow identification of specific components within a sample, such as labeled proteins or DNA. This is especially useful for determining the composition or origin of particles in mixed formulations.

  • Yes. Coriolis applies IFC to BSL-1 viral vectors, including AAVs, supporting the development of gene therapy products through high-resolution imaging of vector-associated particles.

  • IFC is ideal when particle morphology and fluorescence characteristics must be analyzed simultaneously. It is especially useful in early development, formulation screening, or when troubleshooting particle-related issues.

  • Coriolis combines technical expertise with access to a wide range of orthogonal particle characterization tools. This allows us to deliver robust insights tailored to complex biologic formulations and unique client challenges.

Analytical Method Development, Qualification and Validation

For common sample types, we can often apply standardized methods with little setup effort. However, when needed, our experienced analytical experts create or optimize custom methods tailored to your active pharmaceutical ingredient, product type and development phase.

Method Development

Our method development approach tailors sample preparation, method settings and data analysis to the needs of your project and sample.

We include representative samples and, where available, suitable reference standards and stressed/degraded materials, allowing our analytical scientists to design a highly suitable, stability-indicating, robust and repeatable method. Upon request, we will compile a detailed description of the method for your records.

Method Qualification

Method qualification is the initial assessment of an analytical procedure’s performance to show its suitability for its intended purpose.

During method qualification, our analytical scientists perform documented testing demonstrating that the analytical procedure meets criteria in several categories. Criteria may include factors such as repeatability, specificity and robustness. We compile a qualification plan and report, including all relevant data.

Method Validation

Under GMP conditions, method validation confirms that an analytical procedure’s performance suits its intended purpose. Depending on the method’s scope, a broad range of method characteristics, such as specificity, accuracy, precision, limit of detection/limit of quantification (LOD/LOQ), linearity and range, is considered.

During method validation, our analytical scientists perform documented testing demonstrating that the analytical procedure consistently produces a result that meets the predetermined acceptance criteria. We compile a validation plan and report that includes all relevant data.

Depending on the development phase, a fit-for-purpose validation approach can be offered, adjusting the validation required efforts in a phase-appropriate way to meet the method’s needs.

Method Verification

Compendial method verification confirms that a compendial method (e.g., from Ph. Eur. or USP) is suitable and reliable for its intended purpose under the specific conditions of the laboratory.

Unlike full method validation, compendial method verification is often considered a partial validation since the method has already undergone extensive testing and validation during its inclusion in the compendium. The extent of method verification depends on the type of method.

During method verification, our analytical scientists perform documented testing demonstrating that the developed analytical method performs adequately for the specific product or matrix being tested and within the specific laboratory where the method will be employed.

Talk to Our Experts or Request a Quote

Our expert team is ready to answer your questions and guide you to the services best suited to your program’s modality, stage and challenge. If your needs are well-defined, we’ll begin the quotation process.

Description

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Particle Characterization – Overview Analytical ultracentrifugation (SV-AUC / SE-AUC) Asymmetrical flow field-flow fractionation (AF4) Backgrounded membrane imaging (BMI) Dynamic light scattering (DLS) Electrical sensing zone (ESZ) / microfluidic resistive pulse sensing (MRPS) Fourier-transform infrared (FTIR) microscopy Hollow fiber flow-field-flow fractionation (HF5) Light microscopy (LM) Light obscuration (LO) Mass photometry (MP) Micro-flow imaging (MFI) Nanoparticle tracking analysis (NTA) Nephelometry / turbidity Quantitative laser diffraction (qLD) Raman microscopy Resonant mass measurement (RMM) Scanning electron microscopy coupled energy-dispersive x-ray spectroscopy (SEM-EDX) Semi-automated visual inspection (SAVI) Total holographic characterization (THC) Transmission electron microscopy (TEM) Visual inspection

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