Grants & Publications Archives | Expert Cytometry | Flow Cytometry Training Grants & Publications Archives | Expert Cytometry | Flow Cytometry Training


5 Techniques For Dramatic Improvements In Reproducibility

Reproducibility has been defined as “the ability to get the same results using the same experimental system.” This is the minimal standard for which we ought to aim. But ideally, we want replicability, which is the opportunity to obtain the same results using different experimental systems. Repro for Everyone has some guidelines to ensure we meet these minimum standards for reproducibility, but I’m also going to cover 5 aspects of lab operating procedures that can be streamlined and controlled in the interest of enhancing reproducibility. These 5 practices will help your reproducibility as well as saving your time and money. If that isn’t enough, high quality and reproducible experiments might even get you a few more citations due to the ease of following your work.

Read More

3 Compensation Mistakes That Will Ruin Your Flow Cytometry Experiments

Compensation mistakes will ruin your flow cytometry results

Compensation is necessary due to the physics of fluorescence. Basically, compensation is the mathematical process of correcting spectral spillover from a fluorochrome into a secondary detector so that it is possible to identify single positive events in the context of a multidimensional panel. Good compensation requires that your controls tightly adhere to three rules. If the controls don’t meet this criteria, it will lead to faulty compensation resulting in false conclusions and poorly reproducible data. Even among flow cytometry veterans, a strong foundation is occasionally in need of a tune-up. And in a topic as complex as flow cytometry, it’s important that we review the fundamentals on a regular basis. In fact, it is the longtime cytometry expert who must check themselves for any sort of faith in older compensation practices. Science is ever a work in progress, and traditional methods are not always the right methods.

Read More

7 Individual Artifacts In Fluorescence Microscopy And How To Minimize Them

There are 7 different common “artifacts” that may be affecting the quality of your imaging. Before digging into the details, let’s begin by defining an artifact: Essentially, it is any error introduced through sample preparation, the equipment or post-processing methods. This is an important concept to grasp because the effects can cause false positives or negatives, and they can physically distort your data. This is, of course, at odds with your mission to obtain reliable quantitative data. So what can you do to stop these artifacts? The problems can range from dirty objectives to bigger issues like light path aberrations.

Read More

We Tested 5 Major Flow Cytometry SPADE Programs for Speed – Here Are The Results

In the flow cytometry community, SPADE (Spanning-tree Progression Analysis of Density-normalized Events) is a favored algorithm for dealing with highly multidimensional or otherwise complex datasets. Like tSNE, SPADE extracts information across events in your data unsupervised and presents the result in a unique visual format. Given the growing popularity of this kind of algorithm for dealing with complex datasets, we decided to test the SPADE algorithm in 5 software packages, including Cytobank, FCS Express, FlowJo, R, and the original, free software made available by the author of SPADE. Which was the fastest?

Read More

Mass Cytometry Revolves Around These 5 Things

Because mass cytometry allows users to characterize masses so effectively, data can be normalized much more efficiently than what traditional fluorescent flow will permit. If there is no working CyTof at your institution, you can still partner with CyTof-friendly research institutions that have the technology on hand. And because the samples are fixed, you can ship them overnight. This way, they will be analyzed for you. Today’s article will summarize the functionality of mass cytometry technology. This tech has been commercialized largely by Fluidigm in the CyTof systems. There are 5 key points to cover, or takeaways, that cytometrists should keep in mind as they perform their research. The 5 points include how mass cytometry works, panel design, proper sample preparation, data analysis, and imaging mass cytometry.

Read More
Flow Cytometry Education And Consulting - Affordable. Effective. Leading Edge.