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Ask These 7 Questions Before Purchasing A Flow Cytometer

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Written By: Tim Bushnell, Ph.D.

Using funds to make a capital purchase can be an exciting time in a facility. If you don’t have the funds in hand, planning for future purchases requires due diligence to make sure the investment is worth it, and that it will satisfy the needs of the community. That $200,000 or more (sometimes much more) is an investment in the future research capabilities of the facility, so invest wisely.

Over the course of writing instrumentation grants, developing business plans, and acquiring instruments, the following questions should be your go-to checklist — what I look at and what you need answered before spending that hard-won funding.

1. What role does the instrument need to fill?

At the top of the list is identifying the instrument’s role. What does the research community need at the moment, and in the near future? Develop good tracking metrics on current trends in usage, and ask major users about their 1-to-3-year plans. And, don’t forget to investigate funding status.

Your data may suggest you need another cell sorter, but as you investigate gran ...

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6 Microscopy Assays To Determine Cell Health and Improve Your Experimental Results

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Written By: Heather Brown-Harding, Ph.D.

When you’re performing imaging, we always want to make sure that any phenotype we see isn’t just an artifact of unhealthy cells or if you’re doing drug discovery, you want to ensure that the treatment isn’t highly toxic to non-target cells. Therefore, it’s important to understand the health of your cells.

To begin to understand if your cells are healthy you need to ask yourself:

  • Is the cell still dividing?
  • Is the metabolism of the cell changed?
  • Are these features of apoptosis or necrosis?
  • Are the cells dead or on the verge of dying?

Western Blot and flow cytometry are just 2 of the possible methods for answering these questions, but when spatial or temporal information is important, microscopy is best for cell health.

Since cell health is such a crucial aspect to measure, here are 6 types of assays to help you determine the health of your cells using a microscope.

1. Look for morphology loss – or loss of asymmetry.

Use bright field imaging to determine whether your cells are spread out – do they lo ...

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7 Things You Didn’t Know About Imaging Cytometry

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Written By: Tim Bushnell, Ph.D.

We are visual creatures. We seek to capture and describe the world around us. Some of the earliest evidence of this is found in the cave paintings that have been found around the world, like this painting of a horse found in the caves in Lascaux, France.

Figure 1: Cave painting of a horse. From Wiki.

With the development of reliable microscopes, such as those developed by the dutch draper Antonie van Leeuwenhoek. For the first time, we were able to see what we could not see before, probing the unseen and learning in great detail how organisms worked.

Figure 2: Antonie van Leeuwenhoek and his microscope. The figures represent his drawings of red blood cells

Over time, the field of cytometry, that is the analysis of biological processes at the whole-cell level, has exploded and expanded in so many different directions. The flow cytometry can be thought of as a microscope with very poor resolution. The power of flow cytometry lies in its ability to analyze thousands of cells through many dimensions, providing an amazingly detailed understanding of the ...

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4 Steps To Implementing a QC Program For Your Flow Cytometry Experiments

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Written By: Tim Bushnell, Ph.D.

With the emphasis on reproducibility in science, it is important to look at the process of quality control and quality assurance and how these can help improve the reproducibility of data.

First, what is the difference between quality control (QC) and quality assurance (QA)? For the purposes of this article, we will define QC as the activities that are performed to ensure that the product, in this case, flow cytometry data, meets the standard. QA, on the other hand, is focused on how the processes used to develop the product and how these can be improved as well as establishing methods that prevent poor quality product.

A lot of time is spent talking about QC, especially in regards to how the instrumentation performs. Less time is focused on QA. Looking at these two processes in the context of a flow cytometry experiment, QA is focused on the development of the panel, while QC is focused on how well and consistently the panel performs once it is put into production. This blog will focus on one manner of implementing a flow cytometry panel and how to Q ...

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5 FlowJo Hacks To Boost The Quality Of Your Flow Cytometry Analysis

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Written By: Tim Bushnell, Ph.D.

Primary data analysis, that is the analysis at the sample or tube level, is where the populations of interest are identified and the necessary data is extracted for secondary analysis. Since the creation of the FCS standard, flow cytometrists have had the ability to analyze data in third party software because of the communities agreement on the standard. The most recent standard of FCS3.1. The FCS standard divides the file into two components, the listmode file that contains the sequential data from all the detectors. The header file contains what are termed ‘keywords’. These include define keywords that are added to the file automatically, as well as terms that can be defined by the user.

These keywords are only the tip of the iceberg when it comes to performing advanced analytics. This article will focus on the power of FlowJoX (FJX) and provide some tips and tricks to improve the researchers analysis.

1. Embedding and using keywords.

The header information of the FCS file contains a great deal of information that is very useful to review and e ...

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