What is Flow Cytometry Analysis?
Flow Cytometry Analysis is a method to characterize cells at high speed in larger groups that cell biologists implement as a standard method. Recently, relevant features of cells and target macromolecules has become an exciting topic in biotechnology. Typically, cells receive a chemical modification with a fluorescent probe or a dyer. The method allows the identification of cell cycle steps, viability, or biochemical cell kinetics.
Commercial flow Cytometers are new equipment however, for development projects or medical applications it may be necessary to construct flow Cytometers with practical consequences. Here, we will briefly explained some do-it-yourself devices, the flow Cytometers are robustly built using syringe pumps and integrated with 2D and 3D systems.
Integrating syringe pumps with a webcam-based flow cytometer
Rosenauer Vellekoopdevice developed an integrated three-dimensional 3D adjustable optofluidic lens system for forward-scattering/extinction, in this way tracking the biochemical response of different cells is possible. The microfluidic dosage systems are highly relevant in this process. A commercial webcam of 187 frames/sec can take a resolution of 320 × 240 pixels, for the fluorescent excitation a 1 W at 450nm. The free flow cytometer detects rare cells in large volumes at a 500 μL/min (A simplified scheme is show in Figure 1). For that purpose, five Chemyx 200 syringe pumps apply the pressure flow to make the system work with large volumes. The simplicity and low cost of this device suggests that it may have a potential clinical use for mobile health devices. The flow cytometry analysis for resource-poor settings associated with global health is a necessity to be developed (Rosenauer et al., 2011).
Figure 1. The scheme is adapted from Balsam, et al. 2014, the device uses a webcam with a wide-field flow cytometer. In the A panel, the representation shows the modules, a sensor which is the webcam and the lens for amplification, the excitation source, in this case, the laser module, and the flow-cell would need a Chemyx syringe pump to work properly and feed the cell flow. All the information is sent to a computer to collect and analyze the data. The programmable syringe pump is key to keep the hydrodynamic regime on the B panel where the cells are identified by the detection system (Balsam et al., 2014).
Another low-cost flow cytometer based on a webcam capable of cell detection and quantification in a large volume is a 2D flow-cell system which can be designed with a particular hydrodynamic regime. The integration of the webcam and the 2D flow-cell can be achieved with a syringe pump. The flow cytometer technology enables the detection of fluorescent tagged cells in concentrations at low levels as one cell/mL with high flow rates of 500 mL/min. The statistical effectiveness of this fluorescent integrated detection is 100 cells/mL, and 84% are detected which is better for microscopy with ten cells per mL 79% detected. In general terms, the system allows the detection of tagged cells at high volumes (Figure 1)(Balsam et al., 2014).
Flow cytometers are new devices. However, to generate new information and provide evidence for cell biology research it is essential to produce new tools. A way to research with a new flow cytometer is Do-it-yourself cytometers; these devices are robust and provide meaningful data. Chemyx syringe pumps allow the design of modern flow cytometers with better capabilities.
Balsam, J., Bruck, H.A., Rasooly, A., 2014. Webcam-based flow cytometer using wide-field imaging for low cell number detection at high throughput. Analyst 139, 4322–4329. https://doi.org/10.1039/c4an00669k
Rosenauer, M., Buchegger, W., Finoulst, I., Verhaert, P., Vellekoop, M., 2011. Miniaturized flow cytometer with 3D hydrodynamic particle focusing and integrated optical elements applying silicon photodiodes. Microfluid. Nanofluidics 10, 761–771. https://doi.org/10.1007/s10404-010-0707-z