1 – Instrumentation

Getting A New Flow Cytometer? Try Before You Buy (And 2 Other Tips)

By: Tim Bushnell, PhD

There are 4 major ways to sort cells. The first way can use magnetic beads coupled to antibodies and pass the cells through a magnetic field. The labeled cells will stick, and the unlabeled cells will remain in the supernatant. The second way is to use some sort of mechanical force like a flapper or air stream that separates the target cells from the bulk population. The third way is the recently introduced microfluidics sorter, which uses microfluidics channels to isolate the target cells. The last method, which is the most common––based on Fuwyler’s work––is the electrostatic cell sorter. This…

5-Point Guide To Buying A New Microscope For Your Lab

By: Heather Brown-Harding, PhD

Have you ever noticed how painful it can be to purchase a new microscope? It would be hard to miss – this can be a frustrating process. A lot of scientists and students consider the new microscope hunt quite scary for a variety of reasons. It might be that you’re worried you won’t get the right microscope and that you’ll regret it, or you may find that dealing with salespeople, in general, makes you kind of uncomfortable. But remember, salespeople are just human beings like you and me, and if we can treat them as such, the whole process of…

Ask These 7 Questions Before Purchasing A Flow Cytometer

By: Tim Bushnell, PhD

I am still convinced that my first cell sorter was possessed. The number of issues that I had with the system remains hard for me to believe, even after all these years. It had been purchased, in part, from one vendor because the sales rep for a competitor was nowhere to be found. At that time, I admit I wasn’t overly diligent in my research process. Since then, I’ve pinpointed some critical questions that need to be answered before purchasing a new instrument. At the end of the process, a shiny new instrument will arrive at your facility. Make sure…

Instrument Quality Control For Reproducible Flow Cytometry Experiments

By: Tim Bushnell, PhD

The flow cytometer is an integral component of any flow cytometry experiment, and special attention should be paid to ensuring that it is working correctly and consistently. As an end-user, the researcher should be able to sit down at a machine and know that it is performing the same way today as it was yesterday and last week. Equally important is that if any changes in instrument performance have occured, the end-user knows how they have been addressed and corrected, rather than letting them fester and potentially affect the results. Quality control measurements can include a variety of targets, such…

How to Optimize Flow Cytometry Hardware For Rare Event Analysis

By: Tim Bushnell, PhD

Preparing for rare event analysis requires an understanding of the power and limitation of the instrument to be used. From how fast to run the fluidics, to how the signal is processed to the number of gates that can be used in the sorting experiment, each factor impacts the outcome of the experiment.

3 Ways The ZE5 Cell Analyzer Accelerates Flow Cytometry Research Opportunities

By: Tim Bushnell, PhD

Some technological advances are incremental, while others are significant game-changing tools that offer the researcher the ability to significantly improve current assays while allowing for new and novel avenues of research to be performed. With speed, sensitivity, and capacity to spare, the ZE5 fits into the game-changing category. Reduced carryover, increased speed of acquisition, and a large number of parameters all open up new and novel assays while improving the quality and reproducibility of ongoing ones.

3 Advantages Of Using The ZE5 Cell Analyzer

By: Tim Bushnell, PhD

Since the first laser was mounted to create the first flow cytometer, there has been a push for more - more lasers, more detectors, more colors. As a result, today’s researchers require a large number of lasers and detectors to ensure current panels can be run and new, expanded panels can be developed. This can be problematic because, in general, making one decision to improve a cell analyzer can limit the analyzer in other ways. It may seem like an impossible task, but the team of Bio-Rad and Propel Laboratories, collaborated to bring the ZE5™ Cell Analyzer to the market…

3 Advantages FCS Express 6 Has Over Other Flow Cytometry Data Analysis Software Programs

By: Tim Bushnell, PhD

FCS Express is the ideal data analysis software program to use when analyzing your flow cytometry experiments because it is the most user-friendly program available that is both aligned with current data analysis best practices and maintains rigorous quality control standards.

How To Use A Threshold To Reduce Background Noise In Flow Cytometry

By: Tim Bushnell, PhD

Getting a clear signal with reduced noise is an essential component to good data. Adding a threshold when acquiring flow cytometry data is one way to do that. It reduces the number of events by setting a bar that a signal pulse must clear before it is counted as an event. Depending on the importance of the data, the downstream applications for the data (or sorted cells) will dictate how critical the threshold is. In combination with proper sample preparation, appropriate thresholding will reduce debris and ensure best outcome.

How To Set And Monitor Optimal Voltages For A Flow Cytometry Experiment

By: Tim Bushnell, PhD

The best way to take out the fear and agony of setting voltages is to use some optimization methods. The peak 2 method is a useful and robust method of identifying optimal PMT voltage ranges. Refining that to the voltage walk with the actual cells and fluorochromes of interest will further improve sensitivity, which is especially critical for rare cell populations or emergent antigens. This article describes how to set up, monitor, and maintain optimal voltage settings for your flow cytometry experiment.

How To Troubleshoot The Flow Cytometer Fluidics System

By: Tim Bushnell, PhD

Most of the interactions that a user has with a flow cytometer is with the fluidics system, and many of the issues that users will face in troubleshooting problems on the instrument will also be here. Understanding how the fluidics system works on your flow cytometer will help you prevent many common issues, prepare your samples correctly, and protect your data. Here are four important things to consider about the fluidics system in a flow cytometer.

How To Get A Flow Cytometry Job In 5 Steps

By: Tim Bushnell, PhD

Flow cytometry is a powerful technique impacting both clinical and research. When looking for a career, flow cytometry can take you many places. An experienced flow cytometrist can find a job in a biotechnology company, academia, a clinical setting, and more. To be successful in the field, it’s important to seek out new educational opportunities and network with your peers. Here are 5 tips that can help you turn flow cytometry into a successful career.