Articles

Conventional fluorescence microscopy uses fluorophore-labeled antibodies to visualize cellular structures, providing insights into cellular physiological and pathological states. However, the wave nature of light imposes a resolution limit of approximately 200 nm on...

In recent years, technologies for single-cell analysis have evolved rapidly, revealing huge differences between cells once categorized as being the same type. This variability is most apparent within the cells of the immune system, which must undergo multiple dynamic...

The “optimized multicolor immunofluorescence panel” OMIP publication format was launched 11 years ago as a collaborative platform to establish criteria for experimental design, data collection, and analysis [1]. An OMIP is, by definition, a peer-reviewed publication...

A technique that combines flow cytometry with digital microscopy promises novel insights Imaging flow cytometry (IFC) has seen increased uptake in recent years, largely due to instrumentation and software improvements that have made the technique more accessible. By...

The widespread adoption of fluorescence applications is a testament to the importance of fluorescent dyes in biochemical and biological research. The speed, reproducibility, and sensitivity of fluorescence-based analysis methods explain their rapid adoption in all...

Typing “flow cytometry” into the Google search box returns 11,300,000 results! A daunting amount of information to process for anyone approaching the technology for the first time. To help anyone wanting to learn more about flow cytometry navigate this huge body of...

The recent publication of a 43 color panel for the characterization of Peripheral Blood Mononuclear Cells (PBMC) is yet another example of the multiplexing potential of spectral flow cytometry. The technology, first demonstrated by Dr. Paul Robinson at Purdue...

Enhance the quality of your flow cytometry data by using recombinant antibodies Recombinant antibodies have many advantages over conventional antibody reagents. These include exceptional batch-to-batch reproducibility, highly scalable in vitro production, and the...

Breaking through the diffraction barrier for deeper biological insights Super-resolution microscopy (SRM) improves conventional fluorescence microscopy by achieving higher resolution. It enables fine mapping of structures such as microtubule protofilaments,...

Analysis of flow cytometry data requires the identification of a homogenous cell population. This is achieved in a multistep process known as gating. It involves narrowing down the parameters that define the cell population of interest and excluding dead cells and...

From Research to Therapy Flow cytometry is widely used, with many applications in both research and clinical labs. For pharma and biotech labs, the ability to quickly perform multi-parameter quantitative analysis on large numbers of heterogeneous cells makes flow...

Maximize the benefits of spectral flow cytometry by using the right fluorochromes Spectral flow cytometry enables researchers to conduct larger and more comprehensive flow cytometry experiments than ever before, in some cases with over 40 different fluorochromes in a...

Some of the latest developments advancing scientific research With 2021 now well underway, we felt it might be timely to highlight some of the newer technologies available to enhance laboratory workflows. Modern dyes designed to increase panel size have proven hugely...

Moving Beyond Immortalized Cell Lines For decades, immortal cell lines such as HeLa, HEK 293, A549, and Jurkat have provided researchers a useful and inexpensive platform in which to model normal and disease cellular pathways. However, researchers are increasingly...

Improve your flow cytometry sample prep for more reliable results Sample material is precious, so it’s essential that flow cytometry data is both reliable and reproducible. This means optimizing every step of your flow cytometry protocol, beginning with sample prep,...

Introducing FluoroFinder’s Fluorescent Dye Directory Innovations in fluorescent dye technology coupled with increased demand for more efficient research tools have resulted in the development of an incredible amount of new fluorescent dye products in recent years....

Intracellular Flow Cytometry Fundamentals Proteins produced by cells get shuttled around and end up in a variety of locations, whether they are embedded on the cell surface, contained internally within the cell, or secreted outside of the cell. Intracellular flow...

Live-Cell 3D Imaging with Fluorescence Nanoscopy Advances in microscopy techniques now allow for ultra-high-resolution imaging of fluorescently labeled markers within living three-dimensional (3D) cell cultures. This provides researchers with a more accurate spatial...

Antigen Density Antigen density refers to the number of target molecules in a specific cell subset.  Antigens expressed at high levels are densely populated on the cell, making them more readily available for antibody detection.  Understanding antigen density is...

How  cytometry  is  helping researchers  fight  a pandemic Flow cytometry is playing a central role in shaping our understanding of COVID-19 disease and the SARS-CoV-2 virus. Here we highlight how research teams are using flow cytometry to detect SARS-CoV-2, identify...

State of Spectral Flow Cytometry Spectral analyzers are the next generation of multicolor flow cytometers offering unparalleled sensitivity, increased detection range, do-it-all flexibility, and ease of use. Unlike conventional flow cytometers which use dichroic...

Analysis Software   FluoroFinder’s panel builder tool is the obvious choice to simplify your flow cytometry experiment design, but you will also need to choose software for efficient data analysis. Your lab or core facility may already have one or more analysis...

Flow Cytometry Applications for Blood Immunophenotyping Immunophenotyping refers to the process of detecting expressed protein markers to identify and quantify specific subsets of cells within a heterogeneous population.  Flow cytometry is the most common method of...

Tandem Dyes The commercial availability of antibody-linked fluorescent dyes has expanded rapidly over the past decade. Where researchers were once limited to a handful of common dyes (FITC, PE, APC, PerCP, etc.), they can now choose from an extensive list of...

Flow Virometry Multicolor flow cytometry can be used to detect and characterize viral antigens. This technique, called flow virometry, allows researchers to define viral particle profiles and better understand the relationship between infectivity and transmissibility...

Image Flow Cytometry Image flow cytometry (IFC) is a technique that combines fluorescent immunostaining and microfluidics with powerful digital imaging software. Like flow cytometers, image cytometers analyze heterogeneous cell populations across multiple parameters...

The replication crisis, or data irreproducibility crisis, is now a well-established problem across all scientific disciplines. This inability for researchers to replicate published experimental results exposes data integrity issues and risks potentially faulty...

Flow Cytometer/Cell Sorter Instrument Types Flow cytometers and cell sorters are two powerful tools for the accurate and efficient characterization of cells.  Both instrument types measure forward scatter (FSC), side scatter (SSC) and multiple fluorescent parameters...

Advances in flow cytometry instrumentation, technology and chemistries have enabled researchers to expand their experimental parameters rapidly. Experiments that were once limited to the detection of three to four parameters have increased to detect up to 40 compounds...

Fluorescent proteins are an important part of any molecular biologist’s toolkit. Recombinant DNA techniques allow researchers to incorporate a fluorescent protein into an engineered plasmid and track its expression over time with flow cytometry or, more commonly,...

The enzyme-linked immunosorbent assay (ELISA) offers a simple and effective method for rapid biomarker detection.  The main benefits of ELISA include good specificity, low cost, and straightforward readout.  However, the limited sensitivity of standard colorimetric...

Background fluorescence includes any signal detected beyond what is generated by the fluorochromes being measured.  The three major sources of background fluorescence include autofluorescence, spectral overlap, and undesirable antibody binding. Because false signals...

The field of flow cytometry continues to progress toward the goal of collecting and analyzing more data. Here we explore two recent advancements: spectral analyzer platforms to collect more data, and machine learning algorithms to analyze larger datasets. Machine...

Pathologists and life science researchers are increasingly performing multiplexed assays on formalin-fixed, paraffin-embedded samples.  This allows for the collection of more data from a single tissue specimen, far beyond the typical single-color immunohistochemistry...

Compensation is the process of correcting for spillover when one fluorophore is detected in multiple channels.  It is required for most experiments of four or more colors to identify the correct signal that should be measured in each channel.  This quick list can help...

When designing a flow cytometry experiment, it is important to consider the relative brightness of each fluorescent label on your specific instrument.  Generally, it is best to assign brighter fluorochromes to weakly expressed markers, and dimmer labels to strongly...

Flow cytometry can be performed directly using a fluorescently labeled primary antibody, or indirectly using an unconjugated primary antibody with a labeled secondary. Most researchers prefer direct detection due to its simplicity and ease of use. However, certain...

The inability to resolve signals with overlapping color spectra has traditionally limited fluorescence microscopy to a “color-barrier” of 5 colors. However innovative new developments in microscopy techniques and analysis software are allowing researchers to resolve...

Flow cytometry cell cycle analysis typically involves using a DNA binding dye to determine each cell’s total DNA content. Sorting cells by cell cycle phase is then possible, as the total DNA content fluctuates as cells pass through G0/G1, S, and G2/M phases. Cells in...

The first step to any successful flow cytometry data analysis is proper gating. For a complete overview of gating terminology, types and strategies, check out our previous Gating Newsletter. Here we outline 6 questions that cytometry researchers should ask themselves...