Quantum MESF for Intracellular Fluorescence: Telomere Content Measurement
From time to time, we’re asked about whether we have tools to help with quantitation of intracellular markers, probes, etc. To which we respond: "Heck yeah…!”
As cytometers don’t mind whether cellular fluorescence is internal or external, our Quantum systems for fluorescence quantitation are just as useful for analyzing intracellular fluorescence as they are for surface expression analyses. As with surface-labeled cells, the beads in the kit are run on the same day and at the same fluorescence (PMT, compensation) settings as internally-labeled samples. Channel values for beads are entered into our QuickCal template to calculate a regression associating channel value to standardized fluorescence (MESF) units. See the Quantum MESF and QuickCal datasheets and the video below for more information.
There are many types of quantitative intracellular analyses, but as one notable example, Quantum MESF kits are commonly used in flow-FISH (fluorescence in situ hybridization) methods for the determination of telomere length in studies of aging, carcinogenesis, cardiovascular disease, etc. Protocols are provided in the below references:
Kelesidis T, Schmid I. (2017) Assessment of telomere length, phenotype, and DNA content. Curr Protoc Cytom; 79:7.26.23
Wand T, Fang M, Chen C, Hardy N, McCoy Jr JP, Dumitriu B, Young NS, Biancotto A. (2016) Telomere content measurement in human hematopoietic cells: comparative analysis of qPCR and flow-FISH techniques. Cytometry A; 89(10):914-921.
Baerlocher GM, Vulto I, deJong G, Lansdorp PM. (2006) Flow cytometry and FISH to measure the average length of telomeres (flow FISH). Nature Protocols; 1:2365-2376.