My sample (erythrocytes diluted in PBS) only contains negligible amounts of other cell types.
I stumbled across difficulties using the DeNovix CellDrop, as cell count estimates would vary vastly between measurements. Furthermore, I simply could not find any settings allowing for complete detection of erythrocytes - detection rates varied a lot between consecutive measurements (thus influencing total cell count estimates).
Moreover I was not able to obtain any protocols/setting files for the alternative availiable in our Lab - a Casy cell counter. Are there any protocolls/settings availiable for reliable erythrocyte counting with this machine? (Perhaps also allowing for reliable quantification of cell debry?)
I am truly looking forward to reading your constructive comments,
Michael
Visual counting of erythrocytes can be challenging. They are almost colorless and you have to dilute them properly so that they can be separated from each other without too much dilutional inaccuracy. I would also check the discriminators, erythrocytes from different animals have different sizes.
A typical human RBC has a thickness of 2-3 μm, diameter of 6-8 μm, cell volume of 90 fl, and surface area of approximately 136 μm2. Depending on species, RBC shape and size vary. https://www.intechopen.com/books/blood-cell-an-overview-of-studies-in-hematology/measurement-techniques-for-red-blood-cell-deformability-recent-advances
Kindly check https://www.intechopen.com/books/blood-cell-an-overview-of-studies-in-hematology/measurement-techniques-for-red-blood-cell-deformability-recent-advances
Also check https://study.com/academy/lesson/red-blood-cells-important-measurements.html
I should have specified: we exclusively perform ex-vivo measurements on human erythrocytes. As the CellDrop did not perform well enough for regular use, we are now solely looking for empirically tested/experimentally used Casy-Cell counter protocolls/settings/tricks that would enable us to further enhance our precision. (We do have basic knowledge about Casy-Cell counter techniques)
Jukka Salminen: Thank you for your answer!
We think that the dilution - although performed properly (right measurement range) - still overwhelms the capacity of the CellDrop to measure the sheer abundance of erythrocytes. Moreover, shape and "distribution"/tendency to tilt might slightly throw off the CellDrops algorithm.
Chinaza Godswill Awuchi I really appreciate your answer!
As our lab solely focusses on erythrocytes we do have a lot of knowledge about their rheology etc. At the moment we are simply searching for settings/tricks, that have been experimentally used in order to adequatly and precisely measure cell counts.
Hi Michael,
My name is Dan Schieffer, I'm a scientist at DeNovix, the company that makes the CellDrop. I may be able to help you by optimizing the protocols for your cells if you still are looking for a solution.
Dear Mr. Daniel Schieffer - please excuse my delayed response.
I´d really appreciate your help! So far, we managed to establish reliable settings for the Casy - yet, using a second option/trying out a different methodological approach for measuring cell counts would certainly be interesting!
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology