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Tuesday, June 6, 2023

Move Cytometry Coaching: 7 High Suggestions

Move cytometry coaching is essential to make knowledgeable selections and designing your protocols to get good knowledge. Do some background studying on key subjects like fluorescence and hydrodynamic focusing. Know why you need to use the movement cytometer and whether or not your utility is common or area of interest. Take part in sensible coaching to cement your data.

So that you’ve received your movement cytometry coaching booked and are one step nearer to that treasured knowledge.

In our expertise, the scholars who hit the bottom working with their movement cytometry experiments are those who absolutely interact with the coaching program. Those that don’t—properly, let’s simply say their first few movement makes an attempt aren’t a lot enjoyable.

Sometimes, movement cytometry coaching will encompass some idea plus a sensible session to study cytometer operation and knowledge acquisition. At our facility, the format is as follows:

  • ‘Intro-to-flow’ on-line course plus some cytometer operation movies.
  • Coaching session 1: (or, ‘how to not break the cytometer!’).
  • That is run in teams utilizing beads supplied by the power and covers fundamental steps akin to cytometer startup, altering sheath/waste, priming, establishing an experiment within the software program, instrument settings (voltages/good points), compensation, and easy hierarchical gating.
  • Coaching session 2: customers convey their samples and arrange their particular experiment, making use of data and expertise from session 1.
  • Comply with-up help as requested till a consumer feels assured.

Right here’s my recommendation on learn how to get probably the most out of your movement cytometry coaching.

1. Know Why You Need to Use Move Cytometry

Don’t rock as much as coaching with out an concept of what movement cytometry is and the way you need to use it in your individual analysis. You’d be shocked what number of college students flip up as a result of their supervisor informed them to, however don’t know what experiments they are going to carry out.

You’ll have higher questions, and facility workers can be higher in a position to goal the session towards your wants.

Take into consideration:

  • The pattern sort. Will you’re employed with entire blood/PBMCs, digested tissue, or cultured cells? Your facility will be capable of advise you on pattern prep and supply ideas for sample-specific instrument setup.
  • The kind of experiment you’ll carry out. Will or not it’s immunophenotyping or a practical assay akin to cell cycle or transfection?
  • Will you carry out a multi-color experiment, or do you will have a easy panel (mixture of antigens/fluorophores and different fluorescent dyes)? This may increasingly affect the kind of cytometer you undertake coaching on and can assist facility workers determine what different ideas you’ll must study, for instance, compensation.
  • Will you be constructing on a protocol already developed in your lab, or do it’s good to begin from scratch? Remember that designing a brand new movement cytometry experiment from the beginning can take 6 months or extra.

2. Belief the Course of: Do the Pre-work

In case your facility asks you to do some prep work upfront of your coaching—do it! They’ve designed it that manner primarily based on years of expertise. The supposed studying outcomes of the sensible session will construct on the pre-work. You don’t need to get caught out!

In coaching session 1, we ask customers to show cytometer operation, akin to altering sheath and priming. Actions construct on ideas from ‘Intro-to-flow’, for instance, gating on single cells.

If there isn’t a pre-work required, do your homework. We advocate that you simply familiarise your self with the next ideas:

3. Be Ready and Be On Time

Core services are busy locations, and facility workers have tight schedules. In case you are late you’ll miss essential data or received’t have time to finish your experiment set-up. Be sure to know the place to search out the power, and in the event you should put together samples, permit loads of time—it all the time takes longer than you suppose! Even higher, stain them the night time earlier than and repair them.

Discover out in case your movement cytometry coaching will use beads supplied by the power or in case you are anticipated to supply samples. We cut up our coaching into two classes. The primary is a bunch session utilizing beads, and the second is bespoke with consumer samples. This allows the learner to use their expertise from session 1 to arrange their precise experiment.

In case you are requested to organize samples, be sure to know:

  • A cell staining protocol. Ask your facility if no one in your lab has carried out this.
  • What buffer to convey your cells in. That is normally PBS or FACS buffer (PBS plus FBS and EDTA).
  • What tubes to convey your cells in (e.g., ‘FACS’ tubes: 5ml spherical backside tubes, Eppendorfs, and so forth.).
  • Which controls are needed to investigate your knowledge, for instance:
    • Compensation (single shade) controls.
    • Fluorescence minus one (FMO) controls for gating.
    • Unstained management.
    • Constructive/unfavourable experimental controls.
  • Biosafety procedures:
    • Do your samples require fixation?
    • How will you transport them safely?
    • What if there’s a spillage – will you clear it up?

One or two samples plus controls is good for coaching. One thing consultant of your actual experiment, that may be thought-about a observe. Don’t flip up with 50 tubes or these treasured medical samples. The objective of coaching shouldn’t be the experiment itself however to get you assured with establishing the cytometer to accumulate high-quality knowledge.

Don’t neglect to convey any handouts given by the power, or a pocket book/laptop computer for taking notes.

4. Go With the Move

Get caught in and have interaction with actions. For our sensible movement cytometry coaching classes and workshops, we design an exercise to ship every studying final result. These require scholar participation with content material and friends. Experiential studying, or ‘studying by doing’, results in a deep understanding in a technical setting. [1]

For instance, we give customers an inventory of fluorophores that can be used within the coaching experiment and ask them to pick out the right laser/filter. This allows us to evaluate their understanding of this idea, and so that you can determine any gaps in your data. It’s not a check. We simply need to goal additional instruction the place required.

Ask questions! Facility workers love to speak about movement cytometry—so ask them. They’ll even be asking you inquiries to examine that studying is going down. Questioning, dialogue, and explaining facilitate studying. Do not forget that there aren’t any silly questions, movement facility workers have been in the identical place as you at one level.

Actually, it’s a two-way course of. College students typically have a perspective we’ve by no means thought-about earlier than, and it helps us study too.

5. Embrace Collaborative Studying

Social constructivism is the speculation that data is acquired when learners work together to assemble understanding. [2] Determining options to duties with others in your coaching group will end in a deeper understanding than if the trainer merely demonstrates or provides all of the solutions.

One thing you may need missed, your peer would possibly bear in mind, and vice versa. Assist your co-learners out if they’re struggling; explaining ideas in your individual phrases helps you consolidate your studying. Then again, don’t be afraid to ask others in the event you’re undecided.

After now we have launched a couple of options within the acquisition software program, we give much less detailed directions (for instance – show the information in P1 and visualize the entire unfavourable inhabitants) and take a step again. The scholars assist one another bear in mind learn how to change the displayed knowledge and set a biexponential axis (an axis with unfavourable many years, i.e. beneath zero).

We additionally discover that college students carry out higher of their second coaching session in the event that they work with a companion. So buddy up in the event you’re engaged on an analogous challenge as one other new consumer.

The important thing phrase right here is collaboration. It’s not a contest, and you must by no means examine your self to others. Everyone seems to be from completely different backgrounds with completely different ranges of expertise and completely different talent units. Sharing expertise makes us carry out higher.

6. Be Well timed

Don’t guide your movement cytometry coaching session in the event you received’t contact a movement cytometer once more for one more six months. You received’t bear in mind something! Attempt to arrange coaching inside a few weeks of your first experiment in order that your data is recent.

We anticipate that the primary and second coaching classes will happen inside a few weeks, and experiments ought to start throughout the similar month.

In fact, plans change. So in the event you aren’t prepared to start experiments, or it has been a while because the final time you touched a movement cytometer, ask your facility in the event you can have a refresher. On the very least, learn via your notes, refresh your self with the speculation, and produce a observe pattern to get reacquainted. You don’t need to launch straight into an essential experiment and are available away with unusable knowledge.

Don’t neglect, facility workers are there that will help you. Ask in the event you can have an assisted session however be sure you do that upfront—not 5 minutes earlier than your reserving as they’re unlikely to be obtainable.

7. Replicate Don’t Refract

Replicate on what you will have realized and the way it applies to your individual experiment. The cliché goes, you don’t know what you don’t know! Listed here are some key components you must take into account:

  • What fluorescent dyes will you utilize, and are they appropriate with the cytometers obtainable? Will it’s good to change any filters?
  • What is going to your gating technique be? How will you utilize the mix of antigens to determine your inhabitants of curiosity?
  • Are you performing any functions akin to cell cycle or calcium flux which will require a selected instrument setup?
  • What controls will it’s good to analyze your knowledge? Any multi-color experiment would require single-color, and FMO controls at least, however there could also be controls particular to your experimental setup.

Information acquisition is just one step of a movement cytometry experiment. Are you going to want coaching and help on different components, akin to:

  • Pattern prep and marking. The adage in movement cytometry is, ‘Rubbish in, rubbish out’. You want an excellent pattern prep to get good knowledge.
  • Panel design. This implies pairing the antigens with fluorophores to attain the most effective decision between populations
  • Information evaluation:
    • In software program akin to FlowJo or FCS Specific.
    • Excessive parameter knowledge evaluation—creating pipelines with dimensionality discount/clustering algorithms.
    • Information becoming fashions akin to cell cycle or cell proliferation.
    • Compensation—figuring out and stopping compensation errors

Be sure to’re on the power distribution checklist, so that you’re within the loop of any workshops. Join mailing lists such because the Purdue cytometry checklist, and be part of societies such because the Royal Microscopy Society (it’s for movement too!) and your regional membership.

Suggestions for Move Cytometry Coaching Summarized

You’ll get the most effective expertise in your movement cytometry coaching you probably have a transparent concept of your proposed experiment, achieve a sound grasp of the theoretical components, turn out to be conscious of your gaps in data, and have interaction in two-way communication together with your facility. Good luck and have enjoyable!


  1. Kolb DA (2021) Institute for Experiential Studying What’s Experiential Studying? (Accessed 23 March 2023)
  2. Edmunds S and Brown G (2012) Efficient Small Group Studying: AMEE Information No. 48. Med Educate. 32(9):715–26

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