When red blood cells are stimulated by toxic compounds (e.g. mercury) they change morphology to acanthocytes. Most part of papers report SEM analysis; is it possible to apply a flow cytometric method?
Yes, but you'd have to use imaging flow cytometer, such as Amnis Image Stream (which is pretty much like high-throughput microscopy + flow).
Are there papers about? A classic procedure predicts the use of annexin V-FITC?
Thanks a lot
In theory, you might be able to identify your acanthocyte population simply by using Forward and Side scatter, as these cells appear to be smaller and possibly more granular than typical erythrocytes. Have you ever looked at your control and toxin treated cells via SSC/FSC? Happy to have a look, if you could post a picture of your flow cytometry data.
AnnexinV is used to detect phophatidylserine exposure - this is for example the case for apoptotic neutrophils that can be thus phagocytosed by macrophages. I have found one article, where AnnexinV was used to detect changes in the phospholipid structure of erythrocyte membranes (PMID: 8562945), so might indeed be worth using this as a stain.
I think you can only determine the change in cellular size by flow cytometry by looking at Forward scatter and sometimes cellular complexity by side scatter. As far I know morphology can not be determined by this technique.
You cannot directly measure morphology. However, as Balkrishna Chaturvedi mentioned, you can get an idea from light scatter. I caution that the relationships between light scatter and cell size and shape are not always simple, so don't try to over-interpret them, and always compare to standard particles. I believe that has sometimes been used for looking at osmotic changes.
Some instruments (such as the InFlux) also record directly the time it took for cells to pass (trigger pulse width). On the FACS Canto, it's the pulse width (e.g., SSC-W), I think, or you can get the relationship from height to area (area of the pulse is not directly measured, but calculated from pulse height and pulse width of each signal, assuming a normal shape).
These approaches would allow you to track changes in morphology quickly over time or quickly determine the proportions of cells that might differ in morphology (using direct microscopic analysis to see what the morphological change corresponds to)
A final caution is that light scatter may vary simply due to the orientation of a particle as it transits the laser beam.
Yes, the alteration of RBC morphology can be studied by flow cytometry from the scatter parameters. However the instrument setup and appropriate controls of normal discoid and positive control of spheroid RBC is required for analysis. You may follow the link bellow
https://www.researchgate.net/publication/236252002_Quantifying_morphological_alteration_of_RBC_population_from_light_scattering_data?ev=prf_pub
Article Quantifying morphological alteration of RBC population from ...
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