Nov 21, 2020

Public workspaceCELL COUNT- 03 - Automated cell count with Trypan Blue Solution by Cellometer Auto T4 Cell Counter

  • 1Center for Research in Medical Pharmacology, University of Insubria (Varese)
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Protocol CitationMarco Cosentino, Elisa Storelli, Alessandra Luini, Massimiliano LM Legnaro, Emanuela Rasini, Marco Ferrari, Franca Marino 2020. CELL COUNT- 03 - Automated cell count with Trypan Blue Solution by Cellometer Auto T4 Cell Counter. protocols.io https://dx.doi.org/10.17504/protocols.io.bpxtmpnn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 21, 2020
Last Modified: November 21, 2020
Protocol Integer ID: 44755
Abstract
Published work using this protocol:

- Kustrimovic N., Comi C., Magistrelli L., Rasini E., Legnaro M., Bombelli R., Aleksic I., Blandini F., Minafra B., Riboldazzi G., Struchio A., Mauri M., Bono G., Marino F., Cosentino M. Parkinson’s disease patients have a complex phenotypic and functional Th1 bias: cross-sectional studies of CD4+ Th1/Th2/T17 and Treg in drug-naïve and drug-treated patients (2018). Journal of neuroinflammation, 15(1), 205. https://doi.org/10.1186/s12974-018-1248-8


- Kustrimovic, N., Rasini, E., Legnaro, M., Bombelli, R., Aleksic, I., Blandini, F., Comi, C., Mauri, M., Minafra, B., Riboldazzi, G., Sanchez-Guajardo, V., Marino, F., & Cosentino, M. (2016). Dopaminergic Receptors on CD4+ T Naive and Memory Lymphocytes Correlate with Motor Impairment in Patients with Parkinson's Disease. Scientific reports, 6, 33738. https://doi.org/10.1038/srep33738


- Cosentino M., Ferrari M., Kustrimovic N., Rasini E., Marino F. (2015). Influence of dopamine receptor gene polymorphisms on circulating T lymphocytes: A pilot study in healthy subjects. Human immunology, 76, 10, 747-752. https://doi.org/10.1016/j.humimm.2015.09.032


- Boydum A.Isolation of mononuclear cells and granulocytes from human blood. Scand.J.Clin.Lab. Invest. 21 (Suppl.97): 77-89, 1968


- Alex Mabou Tagne, Franca Marino, Massimiliano Legnaro, Alessandra Luini, Barbara Pacchetti and Marco Cosentino. A Novel Standardized Cannabis sativa L. Extract and Its Constituent Cannabidiol Inhibit Human Polymorphonuclear Leukocyte Functions. Int J Mol Sci2019 Apr; 20(8): 1833. Published online 2019 Apr 13. doi: 10.3390/ijms20081833.


- Angela Scanzano, Laura Schembri, Emanuela Rasini, Alessandra Luini, Jessica Dallatorre, Massimiliano Legnaro, Raffaella Bombelli, Terenzio Congiu, Marco Cosentino, Franca Marino. Adrenergic Modulation of Migration, CD11b and CD18 Expression, ROS and interleukin-8 Production by Human Polymorphonuclear Leukocytes. Inflamm Res. 2015 Feb;64(2):127-35. doi: 10.1007/s00011-014-0791-8. Epub 2015 Jan 6.

Materials
Instruments needed:
- Plastic labware

Materials:
Trypan blue catalog number: T8154-100mL Sigma, Italy
Safety warnings
Cancerogenic solution, BE CAREFUL WHEN HANDLING IT!
For automatic cell count with Cellometer machine use Trypan Blue.

The machine will calculate the n°of cells/ml and the % of viability.


Take Amount10 µL of cell suspention and add an equal amount of Trypan Blue. Use all the volume to place it in the given counting chamber. Place the chamber inside Cellometer and count.



Document
SOLUTION- 09 - Trypan Blue solution
NAME

SOLUTION- 09 - Trypan Blue solution

CREATED BY
Farmacologia Medica

Equipment
Cellometer Auto T4
NAME
Automated cell counter
TYPE
Nexcelom Bioscience
BRAND
EuroClone
SKU