Articles

One of the most common complaints when using fluorescently-labeled antibodies for staining biological samples is that the specific signal is too low. Another is that the background is too high, obscuring detection of the target. We spoke with Eric Torres, Ph.D.,...

Recombinant antibodies have become indispensable tools for flow cytometry due to the advantages that they offer over traditional polyclonal and monoclonal antibodies. We spoke with Mathilde de Jong, Ph.D., Product Manager for Flow Antibodies at Miltenyi Biotec, and...

DNA-PAINT (DNA points accumulation for imaging in nanoscale topography) is a super-resolution microscopy technique that exploits the transient binding of fluorescently labeled DNA probes. It has been widely adopted for scientific research owing to its accessibility,...

Flow cytometry requires more controls than most other immunoassay techniques because it must account for more potential sources of variation. In addition to experimental controls, which are used to verify the specificity of any staining and ensure assay consistency,...

It has been almost half a century since Georges Köhler and Cesar Milstein first described hybridoma technology, which is still widely used for producing monoclonal antibodies today. Over the years, the methodology has evolved and alternative approaches have been...

When using established methods for immunofluorescent staining of tissue sections, the need to avoid spectral overlap limits the number of markers that can be detected. This can mean that important information is missed, which is of particular significance when working...

Functional probes are essential tools for flow cytometry, with utility for studying a wide array of cellular processes. Here, we speak with Alexis Madrid, Ph.D., Assistant Director of Bioscience at Biotium and Chris Manning, Associate Director, Flow Cytometry at Cell...

Fluorescence recovery after photobleaching is a technique for characterizing the mobility of cellular molecules   Fluorescence recovery after photobleaching (FRAP), also known as fluorescence redistribution after photobleaching, is a microscopy-based technique...

Cellular senescence has important roles in embryogenesis, tissue repair, and tumor suppression. However, prolonged senescence can be detrimental to human health, contributing to the development of conditions such as cancer and various age-related pathologies....

Two-dimensional (2D) cell culture represents a cornerstone of scientific research due to its relative simplicity and the establishment of standard techniques over time. However, its position is under threat from 3D cell culture for the more physiologically relevant...

Cellular function encompasses the basic processes a cell must undertake to do it’s assigned job. That could include protein production, enzymatic degradation, proliferation, apoptosis, cell signaling, mitochondrial respiration, buffering reactive oxygen species, etc,...

So many new fluorophores have been released in recent years, it can be hard to keep track of how they are all similar or if any are truly novel. Sometimes, a company will create a new brand name for a fluorescent chemistry that is already commonly used which can make...

Multiparameter fluorescence imaging offers many advantages for scientific research. Not only does it use significantly less sample than generating a series of single-analyte images, but it also saves time and can provide a more complete picture of the system in...

Although intracellular flow cytometry evolved more recently than surface staining, it is far from new. Reports dating back almost 50 years describe the use of intracellular flow cytometry to track the cell cycle, through differential staining of DNA and RNA with...

Historically, fluorescence microscopy was limited by the number of parameters that could be imaged on a single sample due the spectral overlap of the fluorophores available at the time. There was a prevailing dogma that only one fluorophore could be assigned to each...

Standardization may not be the most exciting or cutting-edge topic in biomedical research but in an era where we lament the lack of reproducibility and distribute blame to reagents, sample prep and general technical know-how, standardization is something every...

Whether you are using a traditional cytometer that employs compensation and does not exceed 18 colors or a spectral cytometer that employs an unmixing algorithm and has the potential, currently, to analyze 50 color panels, the basic tenets of flow cytometry panel...

In 2004, Perfetto and Roederer published the first paper detailing a 17-color flow cytometry experiment by incorporating a relatively new octagon emission array off the violet 405nm laser (1). The additional laser and 8 additional channels for violet excited...

The selection of method and fluorophores for super-resolution microscopy depends on several considerations.    Fluorescence microscopy is one of the most widely used imaging modalities for scientific research owing to its capacity for multiplexed detection and its...

Stem cells offer vast potential for new medical treatments   With the unique abilities to self-renew and rebuild functional tissues, stem cells have long been of interest to researchers as potential tools for regenerative medicine. 2006 saw a major turning point...

Multimodal spatial profiling at a single-cell level promises to transform scientific research   Single-cell analysis techniques such as fluorescence-activated cell sorting (FACS) and single-cell DNA and RNA sequencing have broadened researchers’ understanding of...

What is Spectral Spillover? In flow cytometry, spectral spillover occurs when the emission of one fluorophore overlaps the optimal detection channel of a different fluorophore. Spillover is a concern in flow cytometry, as it can make fluorescent signatures difficult...

Application-specific validation is critical for reproducible results   Antibodies are essential tools for a broad range of research techniques. However, for data to be both accurate and reproducible, antibodies should be thoroughly validated for each of the...

Aberrant cell proliferation and cell death underlie a multitude of disease states Normal tissue homeostasis depends on a critical balance between cell proliferation and cell death. The cell cycle regulates the former, while the latter occurs via controlled...

Intravital microscopy has vast potential to reveal hidden cellular mechanisms   Intravital microscopy (IVM) is a term used to describe the direct visualization of cells and tissues within a living organism. It was first reported in the 17th century, shortly after...

The use of mass cytometry is growing and includes its adoption for clinical applications Mass cytometry is a variation of flow cytometry that uses antibodies labeled with metal tags rather than fluorophores. Also known as CyTOF® flow cytometry, in deference to...

Fluorescent immunoassays meet different experimental needs   When immunoassays such as western blot and enzyme-linked immunosorbent assay (ELISA) were first reported, they typically produced either a radiometric or chromogenic signal. However, many common...

By measuring intracellular proteins with flow cytometry, researchers can better understand cell signaling and functional responses in conditions of health and disease   Flow cytometry is widely used for identifying different cell types based on the expression of...

Choosing the right fluorophores for microscopy-based research is critical for accurate results Using fluorescent reagents for microscopy-based research offers several advantages. Not only does fluorescent detection enable multiplexing, which can be especially useful...

Common problems can often be addressed by optimizing experimental design   Flow cytometry is a technique for analyzing individual cells in suspension. It works by using a stream of fluid to direct the cells in single file past an interrogation point, where one or...

Viability dyes are critical controls for proper flow analysis. The presence of dead cells can result in unwanted autofluorescence, cell aggregation, non-specific antibody binding, and a compromised capacity for RNA/DNA to proliferate in downstream applications. These...

Fluorescent western blot detection can offer many advantages provided protocols are carefully optimized   While it was once common for researchers to use enzyme-labeled antibodies and chemiluminescent substrates to develop western blots to film, there has been a...

Spectral cytometry offers increased flexibility for fluorophore selection but researchers should still apply best practices for panel design.   It has been almost 20 years since spectral cytometry was first described by the Robinson group at Purdue University...

Protocol optimization is essential for clear, reproducible results   Fluorescence microscopy uses fluorophore-labeled antibodies and other fluorescent reagents to visualize targets of interest in cells and tissues. Experiments can be as simple as detecting one...

  Understanding the biological density of proteins or antigens expressed by each individual cell is an imperative component of all cell-based analytical methods. The comparative change in protein expression, also called antigen density, is indicative of the...

Reducing autofluorescence is critical in fluorescence-based research   Techniques such as fluorescence microscopy, flow cytometry, and western blotting often rely on the use of fluorophore-labeled antibodies. The main reason for this is that fluorescence-based...

Fluorescent cellular analytical technologies allow us to “see” beyond what was historically possible with histological stains or morphological scatter profiles. In the early days, microscopy employed excitation sources like arc lamps, isolating specific wavelengths of...