Airflow and Flow Cytometry Technologies is an advanced laboratory procedure involving the sifting of cells and other biologic and non biologic materials and using lasers to assess and measure their various physical and chemical characteristics as they flow in a liquid medium.
Basically, the lasers interact with the substances in the medium and produce a unique image and/or light emission pattern depending on how the laser actively interacts with the substance.
Thus a unique set of signals is generated, and quite a number of impacts are recorded. This information is subsequently processed for analysis purposes to find out the properties of the particles in the suspension.
How do Airflow and Flow Cytometry Technologies revolutionise cellular analysis, and what innovative applications can we expect in the future?
This flexible device can be employed in a variety of disciplines such as immunology, virology, molecular biology, cancer biology, and surveillance of infectious diseases.
In medicine and diagnostics flow cytometry is broadly used since it is efficient in counting and characterising important cellular populations, for instance, from living tissues such as tumours.
Select a cyclometer and learn its setup.
In the case where there is access to more than one flow cyclometer available in a core facility, it is advisable to speak with the core manager regarding what instruments would be ideal for the research that you intend to perform.
After deciding on the cyclometer, one must learn the features of the instrument and how its configuration would influence the design of the experiment.
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Make a Multicoloured Experiment Panel for Yourself
Your panel must be equipped with antibodies against all markers of interest alongside the appropriate fluoroscopes within your cyclometer setup.
For cells with lower antigen expression, brighter fluoroscopes are more appropriate, while for antigens that are highly expressed dimmer fluoroscopes are more suitable.
More details need to be considered and care taken to avoid “spillover” or cross detection of fluorescence signals in the channels.
This is where our library of over 500,000 items as well as 550 Mercurochrome from 60 leading manufacturers comes in handy by allowing the user to create, store and fine tune multicoloured flow cytometry panels.
In addition, built-in Spectra Viewers and spillover tips machined to specific designs assist participants in unravelling the maze that is known as panel designing.
Enhance your protocol for staining
It is essential to accurately tit rate all antibodies used in an experiment, as their volume should be appropriate for the specific experiment in question.
Be advised, working concentration ranges recommended for an individual antibody within a panel may not apply when other antibodies are present.
Moreover, adequate blocking serums must be applied to prevent non-specific antibody binding.
Staining some extracellular antigens may also prove a challenge. Also and depending on the markers being looked at, fixative and or destabilization’s solutions may be needed. Also be advised that fixation will kill your cells!
Choose your controls
Controls are the most essential tools while designing any flow cytometry experiment. Based on the experiment, viability dyes, FMO, beads, isotope controls, or any others might be used.
It is essential to carry out fluorescence minus one to determine spillover into each channel.
This is particularly valid whenever there is an assessment of populations that are dim or indiscreet.
For additional details about the flow cytometry controls, please read our piece regarding Common Errors in Panel Design.
In addition, think about using a dump channel where you put all your viability dyes and other controls.
Get your sample ready
Under prepared samples will yield incorrect readings and erroneous outcomes. All live cells must be suspended properly to eliminate clumps and stickiness and then sieved to separate dead cell debris which causes auto fluorescence.
Naturally, it is also essential to handle cells with care to prevent death in the course of the experiment. For more ideas on how to prepare samples, you may refer to our Essential Pre-Sort Checklist.
Conduct your tests
The machine time on flow cyclometers is often extremely costly, hence, ensuring when heading to your flow core, your samples will be ready to run in an efficient manner.
In addition, when reserving a cyclometer to use, always remember to include time for preparing for the experiment before and cleaning the instrument after.
Practice these suggestions for Getting More Out of Your Flow Core to make the most flow core time more efficient.
Perform data analysis
Data from flow cytometry is analysed on computer systems employing advanced algorithms.
A large number of both free and paid software solutions are available that come with special features for data editing, representation and sharing.
Some of those analysis software’s capabilities even include performing automatic compensation for fluoroscope spillover.
To see more of such software available, see our Flow Cytometry Analysis Software List.
FAQ’s
What is the crucial part of flow cytometry?
Flow cytometry is a laboratory technique to define the characteristics of suspended cells or particles by analyzing laser beams.
The technique is broad in its application in clinical setups in evaluating the condition of bone marrow, peripheral blood, and other body fluids.
Which application of flow cytometry is most predominantly utilised?
The most frequently conducted procedure on the cyclometer is known as immunodeficient.
This method in fact enumerates and characterises cell populations in a complex mixture which is usually blood, bone marrow or lymph.
What is the Principal use of flow cytometry?
Microscopy, and more specifically flow cytometry, has as its main objective the possibility of counting accurately all or specific types of cells in the sample provided.
In this way flow cytometry is able to provide statistical data about certain cell populations enclosed in a heterogeneous mixture.
What is the measurement unit of turbidity in flow cytometry?
Turbidity measurement is performed in soda centrifuge at low speed 5, 000 rpm for 10 minutes.
What are the benefits of using flow cytometry?
It enables the assessment of thousands of cells in a few seconds in a rapidly flowing concentrated single cell suspension.
This technique is employed in all the life sciences for simultaneous characterisation of populations of cells by many parameters, while keeping the specimen untouched.
CONCLUSION
Airflow and Flow Cytometry Technologies have presented the theory behind flow cytometry and its practical use. Flow cytometry is being applied more and more not only in basic research but also in clinical laboratory science.
For example, there are additional flow cliometric applications such as DNA analysis, assessment of paroxysmal nocturnal haemoglobin (PNH), reticulated counts, and study and diagnosis of acute amyloid leukemics.
In addition to small bench-top flow cytometry units, currently available haematology cell counters also incorporate flow techniques into their group cell analysis.
