4 Fluidics Tips That Will Change Your Flow Data For The Better
Friday is the 4th of July in the US – and we celebrate that day with picnics, spending time OUTSIDE the lab and fireworks. And our Independence, but I’m not up for a political discussion right now. For flow geeks, fireworks are like flow cytometry – they happen in the dark, they are full of many bright colors, and we’re all looking for the patterns the colors make. So in honor of a day outside the lab, it seemed appropriate to talk about fluidics… going with the flow for best results.
A flow cytometer has three major components – fluidics, electronics and optics. From setting the run speed (‘flow rate’), to cleaning the instrument after a run, to changing the sheath fluids, the typical researcher interacts mainly with the fluidics side of the system.
The majority of flow cytometers on the market use a differential pressure to move the fluids and cells around the system. In these systems, the pressure of the sheath fluid sets the speed of the flow. The low, medium and high buttons on the instrument change the differential pressure between... Read More
Counting Cells Will Save Your Flow Cytometry Experiment, If You Do It Like This
Fast, accurate or efficient – pick two. How to decide when you can’t have it all.
The hemocytometer is considered the gold standard for cell counting. Invented by Louis Charles Malassez, this precision etched microscope slide can allow the researcher to count their cells under the microscope with amazing accuracy. It is inexpensive, relative to other methods, but is by no means the most efficient or fast method out there.
The single biggest key to using a hemocytometer is training, training and more training. Since the investigator is visually inspecting the cells within a boundary, the rules of what cells to count and what to exclude on those boundaries becomes critical.
If counting more than one sample, proper cleaning of the hemocytometer is a second critical step. If the cleaning solution is not removed completely, it can cause cell lysis... Read More
Top Flow Cytometer
What is the top flow cytometer? The easy answer is the flow cytometer that matches your needs and fits within your budget. However, before running off to spend cash, consider the following.
What are the current needs of the users? Evaluating the users needs will help define the parameters needed for the flow cytometer. This will include what excitation sources should be available, how many detectors are needed, including specialized detectors (small particle detectors), as the need for automatic sampling. All these factors will in weigh in on the
What are the projected future needs of the users? This is where the crystal ball comes out. Predicting what the future needs of the userbase is a difficult task. The best thing is to evaluate if the cytometer of interest is future proof? That is can it be expanded in capacity and capabilities as the research needs change.
Will the instrument be needed for clinical samples? This... Read More
Flow Cytometry Education
As with all complex technology there are many different levels of education that users should avail themselves of.
Basic instrument operation – this level of education is akin to learning how to drive a car. At this level, the focus will be on how to put the sample on the instrument, how to adjust voltage and collect data. Specific policies of the facility should also be covered.
Advanced instrument operation – at this level of education, the focus should be on advanced techniques on the instrument. This would cover basic troubleshooting – identifying problems and how to solve them. Advanced training should prepare the investigator to independently operate the instrument when the staff are not around.
Data analysis – in parallel with learning how to operate an instrument to acquire data, learning the software to analyze the data is essential. This software specific training should... Read More
Cell proliferation can readily be measured by flow cytometry. Depending on the research question, there are several different techniques that can be used.
Cell counting experiments – this relatively straightforward experiment where the investigator uses one of several counting techniques to see if there is an increase in the populations. This can be done using a microscope and a hemacytometer, a flow cytometer with counting beads, image based tools (like the T-20, the Countess, or the Cellometer), or coulter counter type instruments (including the Coulter Counter the Scepter and the Casy counter)
Cell cycle experiments – one of the earliest techniques developed in flow cytometry. Using a DNA binding dye, flow cytometry can readily identify the major phases of the cell cycle. This technique can be used in comparing samples treated with different conditions to compare how quickly cells move through the cell cycle, or if the cells... Read More