With 2021 now well underway, we felt it might be timely to highlight some of the newer technologies available to enhance laboratory workflows.

Posted Apr 1, 2021

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 moving away from immortalized cell lines in favor of more biologically accurate primary cell models. Here we examine the reasons for this trend and its implications for flow cytometry analysis.

Posted Mar 1, 2021

Whatever cell type you’re studying, our top tips for sample prep will help you get the most out of your flow cytometry experiments.

Posted Feb 1, 2021

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 cytometry refers to the process of staining and analyzing proteins localized inside a cell. In this newsletter, we will go over the basics of intracellular flow cytometry, some important considerations, and how FluoroFinder can help you set up your intracellular flow cytometry experiments.

Posted Dec 1, 2020

Advances in microscopy techniques now allow for ultra-high-resolution imaging of fluorescently labelled markers within living three-dimensional (3D) cell cultures. This provides researchers a more accurate spatial visualization of cell-to-cell interactions, sub-cellular structures, and protein assemblies without significantly affecting cell viability. Here we review the history of and applications for fluorescence-based 3D live-cell imaging…

Posted Nov 1, 2020

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 important for any cell-based experiment using fluorescent antibody detection, as markers with high density will produce a stronger (brighter) signal and should therefore be paired with dimmer fluorophores…

Posted Oct 1, 2020

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 how the virus interacts with host cells, and understand how the immune system responds to the infection….

Posted Sep 1, 2020

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 mirrors and bandpass filters to detect discrete ranges of wavelengths on individual photomultiplier tubes, spectral analyzers incorporate different optical approaches to achieve…

Posted Aug 1, 2020

FluoroFinder’s panel builder tool is the obvious choice to simplify your flow cytometry experiment design, but you will also need to choose a software for efficient data analysis.
Your lab or core facility may already have one or more analysis software packages installed into their flow cytometry workflow. However, not all software are created equally…

Posted July 1, 2020

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 immunophenotyping suspended cell samples and is particularly useful for analyzing whole peripheral blood (WPB) samples for a variety of diseases…

Posted June 1, 2020

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 fluorescent dye lines – spanning the light spectrum from IR to UV. Additionally, researchers can now…

Posted May 1, 2020

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 at the level of individual virions [PMID: 23925291]. Here we examine the progression of flow virometry and some of its most common uses today.

Posted April 1, 2020

Image flow cytometry (IFC) is a technique that combines fluorescent immunostaining and microfluidics with powerful digital imaging software. IFC has become a common technique for a wide variety of research applications. Here we examine the main reasons for, and potential solutions to, IFC’s limited use in clinical and diagnostic applications.

Posted March 1, 2020

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 conclusions entering the collective scientific knowledgebase. Here we explore a few of the organizations committed to promoting open science and some of the data sharing tools developed to address the replication crisis.

Posted February 1, 2020

As you and your lab continue to progress into 2020, discover what was trending in 2019 based on our review. Find out what fluorochromes, cytometers, clones etc. made it to the top of our list… We can’t wait to see what is in store for 2020!

Posted January 6, 2020

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 to detect specific antibody bound markers within a heterogeneous cell population.

Posted December 10 , 2019

As advanced fluorescence detection creates added complexities in experimental design, it is important to step back and understand the fluorescent molecules used in your flow cytometry experiments…

Posted November 6 , 2019

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, fluorescent microscopy methods. Here we explore the history, advantages, common uses and wide selection of fluorescent proteins.

Posted October 7 , 2019

Background fluorescence includes any signal detected beyond what is generated by the fluorochromes being measured.  Here we will explore the causes of and best methods for minimizing or compensating for each of the three background fluorescence sources…

Posted September 3, 2019

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 (IHC) stain…

Posted September 3, 2019

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.

Posted Aug 3, 2019

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 (IHC) stain.

Posted July 9, 2019

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. We’ve developed a quick list of Do’s and Don’ts to help new flow cytometry researchers navigate the basics of compensation.

Posted June 4, 2019

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 expressed markers.

Posted May 7, 2019

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 more biomarkers than previously possible.

Posted April 5, 2019

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 the simplicity and ease-of-use. However, certain fluorescent antibody experiments may require indirect detection with a labeled secondary…

Posted April 5, 2019

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…

Posted March 5, 2019

The first step to any successful flow cytometry data analysis is proper gating.Here we outline 6 questions that cytometry researchers should ask themselves to help improve your gating techniques.

Posted February 5, 2019

We’ve analyzed millions of data points over the past year to identify trends and opportunities in panel design. Find out if your lab is on the cutting edge or falling behind in our 2018 Trends in Fluorescent Experiment Design report.

Posted January 3, 2019

Explore the use of flow cytometry in apoptosis research and examine various fluorescent apoptosis markers.

Posted December 5, 2018

This month we explore how incorporating additional fluorochromes into your experiment can provide valuable data without making your panel design unnecessarily complex.

Posted November 2, 2018

This month we explain why antigen density is important to flow cytometry studies, describe how it is measured, and list some useful references for known antigen expression densities…

Posted October 2, 2018

Getting started in flow cytometry may seem overwhelming, so we’ve broken the flow cytometry experiment down into seven basic steps…

Posted September 4, 2018

Did you know that Flow Cytometry was originally called ‘pulse spectro-cytophotometry’ or that the first flow cytometer was built to detect WMDs? Learn more by reading the full article…

Posted August 7, 2018

Researchers can now use several tandem dyes to improve the fluorescent signal for their experiments. However, while tandem dyes can serve as powerful research tools, they can also introduce a new set of problems to consider. Here we explore the benefits of tandem dyes and review some of the difficulties that they present.

Posted July 10, 2018

Does compensation leave you feeling incompetent?

Check out this “comprehensive” list of compensation guidelines and best practices, gathered from our from our flow community, to help improve your analysis.

Posted June 5, 2018

The “reproducibility crisis”, or the inability for researchers reproduce findings, remains a growing concern for a wide range of scientific disciplines. With this article, we aim to highlight the various efforts being taken by some of our partner reagent suppliers to help combat irreproducible experimental results.

Posted May 4, 2018

You put a lot of time and effort into your flow cytometry experiments. Follow these 7 tips to ensure that your flow data is high-quality and more likely to be published!

Posted Apr 3, 2018

OMIPs help simplify panel design by serving as a useful starting point for new research. In this article, we review the status of the 44 currently approved OMIPs and explain how new OMIPs are being developed with the help of FluoroFinder’s panel builder tool.

Posted Mar 6, 2018

This month we explore the value of using dump channels and explain how they are best utilized to improve your flow data.

Posted Feb 6, 2018

This guide helps to explain the various types of viability dyes, explore their respective benefits, and summarize the commercially available dyes for each group.

Posted Jan 9, 2018

Find out if your lab is on the cutting edge or falling behind the times in our 2017 Trends in Fluorescent Experiment Design report.

Posted December 12, 2017

Flow cytometry gating can often seem like a daunting task. While there is no single solution, experienced cytometrists can recommend several tips beyond “praying for good data”. Here we define gating, explore gating methodologies, and provide some useful tips for properly gating your cells.

Posted November 14, 2017

Flow cytometry research is continually trending toward larger panels with more colors.  This month we explore navigating the ever expanding selection of commercially available fluorescent antibodies and how researchers are getting better data by designing more colorful panels.

Posted October 10, 2017

FluoroFinder has now integrated cytometer configurations for over 500 cores! With so many researchers using FluoroFinder to design their flow cytometry experiments, we wanted to share some of the excellent feedback we have received.

Posted September 12, 2017

Aside from costing your lab time and grant money, shared cytometers are a valuable resource so booking additional time may be difficult. Therefore, we have compiled this list of tips for maximizing your productivity during your time at the flow core.

Posted August 15, 2017

This resource page compiles information on the various commercially available cytometer platforms, including basic features and user reviews, to help you choose the best cytometer for your needs.

Posted July 11, 2017

FluoroFinder recognizes the importance of collaboration for good experimental design, and we are helping to fight the reproducibility crisis by maximizing the available information for each antibody in our database.

Posted June 13, 2017

We have compiled tips to void the 6 most common pitfalls of cytometry experiment design and analysis. Learn how to avoid failed experiments, save time and frustration and get better data!

Posted May 9, 2017

Flow cytometry can often seem complicated and intimidating. Whether you are new to flow or an experienced researcher considering a new technique, we have compiled a glossary of important terms to help!

Posted April 10, 2017

Immunobeads are useful for a variety of flow cytometry related applications. Here we outline the most commonly used bead types and explain how they can be used for cytometer calibration, compensation determination and even cell sorting.

Posted March 7, 2017

Is your lab on the cutting edge or falling behind the curve? We have analyzed thousands of panels to find these interesting trends in fluorescent experiment design.

Posted February 10, 2017

Are you planning to run a cell sort on a precious sample? Concerned about recovery? Be sure to review our pre-sort checklist of 7 things experienced cytometrists recommend to improve cell recovery.

Posted January 10, 2017

Are you designing multicolor panels around your fluorescent proteins? Would seeing your fluorescent proteins and reagents in a multi-vendor spectra viewer help you better anticipate issues?

Posted December 8, 2016

Designing a multicolor fluorescent panel can take months to develop and optimize. Fortunately, Optimized Multicolor Immunofluorescence Panels(OMIPs) have greatly reduced this process.

Posted November 8, 2016

Could a new product improve your experiment? Has that new antibody been vetted by reliable testing? Linking reagent publication histories to antibodies data gives researchers more reliable info during experiment design!

Posted September 8, 2016

Viability dyes enable you to distinguish between live and dead cell populations in your analysis. Learn why this control is critical to most flow cytometry experiments.

Posted June 30, 2016