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4 Ways To Analyze Tissues By Flow Cytometry

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Written By: Tim Bushnell, Ph.D.

Why are we interested in cytometry? What are its practical applications? Let’s start by defining cytometry as the measurement of cellular processes at the whole-cell level. This definition is useful because it includes not only flow cytometry, but any technique that measures at the level of the whole cell. Microscopy, for instance, is a great example of cytometry.

But, why measure in tissues? When flow cytometry is practiced, the cells are broken up. Therefore, any cellular interactions within the sample are also broken up. This includes cell-to-cell contact and virtually any information about the microenvironment. As we continue to discover, the microenvironment can play a dramatic role in cell development, influencing how cells grow and change.

Here are 4 ways to analyze tissues by flow cytometry…

1. Multi-photon cytometry

Multi-photon microscopy doesn’t rely on traditional fluorescent excitation. In the traditional process, a single photon will excite the molecule. Following this, there is a fall back to the ground state along with the de ...

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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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