Flow Cytometry Analysis of Primary Cell Types: 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 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
Tips and Tools for Flow Cytometry Sample Prep Success
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
Introducing FluoroFinder's Fluorescent Dye Directory
To help researchers find the right dyes for their experiments, FluoroFinder is rolling out a new resource to our community: our Fluorescent Dye Directory. We have compiled vital information on hundreds of fluorochromes, fluorescent dye products, and fluorescent proteins into a single Fluorescent Dye Directory to allow researchers to have one place to search for the best fluorescent product for their experiment.
Posted Jan 15, 2021
2020 Trends in Experimental Design
2020 was marked as one of the most significant years for advancements in antibody research within the last decade. As you and your lab continue to progress into 2021, discover what is trending based on FluoroFinder data and feedback.
Posted Jan 1, 2021
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 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
Live-Cell 3D Imaging with Fluorescence Nanoscopy
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: Why Brighter Is Not Always Better
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 Fights COVID-19
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
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 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
Considerations for Choosing a Flow Cytometry 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 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
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 immunophenotyping suspended cell samples and is particularly useful for analyzing whole peripheral blood (WPB) samples for a variety of diseases…
Posted June 1, 2020
Life is Brighter with a Buddy-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 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
The Potential and Limitations of Image Flow Cytometry
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
Fighting the Replication Crisis with Open Research
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
2019 Review: Trends in Experiment Design
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
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 to detect specific antibody bound markers within a heterogeneous cell population.
Posted December 10 , 2019
Understanding Fluorochromes for Flow Cytometry
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
Fluorescent Protein Expression
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
FLISA vs. Fluorescent ELISA
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
Advances in Flow Cytometry: AI and Spectral Cytometry
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
Getting More From Less With Multiplex IHC
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
The Do's and Don'ts of Compensation in Flow Cytometry
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
Using the Stain Index to Design Better Panels
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
Breaking the Fluorescent Microscopy Color Barrier
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
Matching Secondary Antibodies for Indirect Flow Cytometry
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
A Guide to Flow Cytometry Cell Cycle Analysis
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
Six Simple Questions to Improve your Gating Technique
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
2018 Trends on Panel Design
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
Are you getting the most out of your cytometer?
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
Antigen Density Explained
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
7 Steps of a Successful Flow Cytometry Experiment
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
18 Facts About the History of Flow Cytometry
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
There's Nothing to FRET about Tandem Dyes
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
Comprehensive Guide to Compensation
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
Join the Fight Against the Reproducibility Crisis
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
7 Tips to Help Get Your Flow Data Published
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: Stop Reinventing the Wheel!
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
A Guide to Choosing the Right Viability Dye
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
Analysis of Trends in Fluorescent Experiment Design
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
The Art of Flow Cytometry Gating
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
Expand Your Experimental Palette
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
Report: Best Practices in Panel Design
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
Get More From Your Flow Core
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
Cytometer Selection Made Easy
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
Addressing the Reproducibility Crisis Through Better Experiment Design
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
Avoiding Common Mistakes in
Panel Design & Analysis
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 Dictionary
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
Your Guide to Understanding Flow Cytometry Beads
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
Report: 2016 Trends in Panel Design
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
Your Essential Pre-Sort Checklist
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
Tracking Fluorescent Proteins
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
Jumpstart Your Panel Design with OMIPs
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
Designing Better Panels with Antibody Validation Data
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 – A Control You Can’t Live Without
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