The cytometric method used in the analysis of different intracellular molecules, utilizes intracellular staining protocols. To label intracellular antigens, the cells are fixed and then permeabilized before adding a detection antibody. This fixation/permeabilization procedure allows the antibody to enter the cell through the cell membrane without losing its morphological characteristics.
Analysis using cationic lipophilic dyes such as JC-1, TMRE (tetramethyl rhodamine ethyl ester), DiOC6 (3), DiIC1 (5), CMXRos (MitoTracker Red), LDS-751, and rhodamine 123, which localize themselves across the inner mitochondrial membrane. Loss of the membrane potential is detected as a decrease in membrane staining or the fluorescence intensity, as the transmembrane potential decreases.
Cell cycle analysis is a very popular application of flow cytometry. Utilization of the DNA-specific staining allows you to determine a DNA profile, thus e.g. the percentage of cell populations in phases G0 / G1, S, and G2 / M of a cell cycle. This information may be useful, for example, to monitor the effects…
Isolation of cells’ subpopulations from cell cultures or tissues by FACS (fluorescence-activated cell sorting).
Cell viability is measured as the percentage of living, healthy cells in a population. Cell viability tests are used to determine the general health status of cells, to optimize cell culture or experimental conditions, and to measure cell survival after treatment with tested compounds (e.g. in the drug screening process). Apoptosis tests enable the detection…
Cytokine detection using bead-based multiplex immunoassays and BD CBA (Cytometric Bead Array) technology as well as BioLegend LEGENDplex kits.
Cytometric method that is suitable for the intracellular signalling pathways analysis. It allows to distinguish different cell types based on surface markers and at the same time enables analysis of signal protein phosphorylation states.
Cytometric data analysis using various programs: FlowJo – allows the analysis and visualization of cytometric data, an additional algorithm tSNE enables the graphic representation of heterogeneous cell’s populations based on multi-parameter cytometric detection, ModFit- allows to analyse the cell cycle and differentiate cells’ populations based on phases.