BD Accuri News
David W. Galbraith, PhD, is Professor in the School of Plant Sciences and the BIO5 Institute for Collaborative Bioscience at the University of Arizona. On February 1, he will discuss plant genome analysis at our webinar, and is co-author of the related white paper.
Q: Tell us about the research program in which you are using the BD Accuri™ C6.
Dr. Galbraith: One theme behind all of my research for the last thirty years is the analysis of plants using flow cytometry and cell sorting. We developed technologies that are now used all around the world, such as looking at plant genomes by releasing nuclei from plants using a simple chopping procedure and then measuring their DNA fluorescence with a flow cytometer. I've always had in the back of my mind to measure the genome sizes of as many plant species as possible. We don't even know how many plant species there are—we estimate maybe 600,000 across the world, which is a large number! And we know genome sizes for only about 2% of all known plant species.
It's a Noah's ark approach to science now, driven by the fact that humans are altering the landscape and driving many species into extinction. We risk the chance of losing species before we've described them. We're measuring genome sizes as a first step toward cataloguing all of these plant species. The next step would be DNA sequencing of their genomes.
Q: Why did you choose the BD Accuri C6? What features are important to you?
Dr. Galbraith: The simplicity is great. It's essentially portable. There are very few user adjustments, so it's unambiguous. You just put the sample on, and off you go.
It also has a very large dynamic range. Flowering plant genomes span a remarkable range—about a 4,000-fold difference across species. The BD Accuri C6 can accommodate that information on a single histogram. We demonstrated that it was possible to use propidium iodide on the BD Accuri C6 to measure genome sizes across a very broad range.
Q: What does BD Accuri's motto, "Flow Cytometry Within Reach®," mean to you?
Dr. Galbraith: Scientists tend to view flow cytometry with an aura of mystique—that it's a complicated process, like X-ray crystallography or NMR or mass spectrometry, that belongs in a core facility. They think it's tricky to work with and you need a sophisticated understanding. Although I wouldn't argue with the need for sophisticated understanding, the instrumentation has become a lot simpler to operate. Many core facility applications now can be transferred to the individual lab.
Studying Plant DNA and Ploidy with the BD Accuri C6 Flow Cytometer
A fundamental parameter in the biology of any plant is its haploid nuclear DNA content (C-value). Known C-values of higher plants vary from 0.1 pg (Fragaria viridis) to 127.4 pg (Fritillaria assyriaca), or more than three decades. This remarkable range, along with mitotic variation, endoreduplication, and the task of identifying cell nuclei within a background of autofluorescent cellular debris, can make it difficult to determine plant C-values. As a result, C-values are available only for about 2% of angiosperm species.
Two factors make the BD Accuri C6 ideal for determining the nuclear DNA contents of plants. First, its sophisticated digital signal processing system provides an exceptional six-decade dynamic range that encompasses almost the entire known span of plant C-values. Second, the BD Accuri C6's red and blue lasers optimally excite the DNA fluorochrome propidium iodide (PI). A high correlation between PI-DNA emission in the FL2 and FL3 channels places the nuclear events in a narrow diagonal region from which it is easy to exclude fragments and debris.
Webinar: Using Flow Cytometry to Speed Determination of Eukaryotic Genome Sizes (C-values) and Cell Type-specific Gene Expression
Join Dr. David Galbraith, Professor, University of Arizona, and Dr. Charles Cannon, Professor, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, who will describe how they use the BD Accuri C6 in their research on plant biology. The applications discussed will include analysis of plant genome size, quantification of reporter and reference plasmid expression, and a homogenation method for preparing plant cells for flow cytometric analysis. This webinar is cosponsored by BD Biosciences and Genetic Engineering News.
White Paper: Using the BD Accuri™ C6 Flow Cytometer for Rapid and Accurate Analysis of the Nuclear DNA Contents of Flowering Plants
BD™ DNA QC Particles on the BD Accuri C6
Checking resolution with BD™ DNA QC Particles
You can use the BD DNA QC Particles kit (Cat. No. 349523) to validate the performance of your BD Accuri C6 before analyzing DNA test samples. Chicken erythrocyte nuclei (CEN)—in singlets, doublets, triplets, and larger aggregates—are used to check instrument linearity and resolution, while calf thymocyte nuclei (CTN) help to assess doublet discrimination and resolution for a cycling population. Propidium iodide (PI) is included to stain the CEN and CTN DNA.
Webinar—Multicolor Flow Cytometry: Setup and Optimization on the BD Accuri™ C6 Flow Cytometer
Date: Wednesday, February 15
Time: 11 AM PST/2 PM EST
Join BD Accuri scientist Clare Rogers for a look at how multicolor flow cytometry can be used on the BD Accuri C6 to analyze multiple cellular parameters simultaneously. This webinar, a reprise of a popular webinar from last September, will focus on key principles of multicolor panel design, instrument setup, sample preparation, and assay optimization as they apply to the BD Accuri C6. Using a complex blood immunophenotyping application as an example, the talk will also highlight online tools such as the new BD FACSelect™ Multicolor Panel Designer.
New on the Website: BD Accuri Applications
The BD Accuri C6 has proven useful in a broad range of flow cytometry applications, and our new Applications page surveys several of the most prominent. You'll find usage notes, sample data, and links to relevant BD Biosciences resources and reagents.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
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