- Sony Biotechnology
- Sony Biotechnology
Much of cell research happens in the context of cultured or modified cells. More biologically relevant experiments can be done with blood. However, blood cannot entirely capture what takes place within a live organism. In their paper Futamura, et al.1 describes a method to study the movement and subsequent interactions of immune cells in live mice by flow cytometry.
To study cell movement, KikGR knock-in mice were used.2 KikGR mice express the photoconvertible fluorescent protein KikGR in a cell-specific manner. KikGR fluorescence changes irreversibly from green to red upon exposure to violet light. In these experiments, inguinal lymph node (LN) of KikGR mice were exposed to violet light for photo-conversion of KikGR. All of the cells in the inguinal LN were subsequently labeled with KikGR-Red signal. Twenty-four hours after photo-conversion, the cell migration from the photo-converted LN to other anatomical organs and the subsequent replenishment of cells in photo-converted LN was studied with an 11-color panel using an SP6800 spectral cytometer.
With the SP6800, phenotyping the T and B cell subsets of the LN was easily obtained with spectral unmixing without the usual complexities of color compensation. Furthermore, cellular autofluorescence of the various cell types assisted in phenotyping the subsets. Read more about this research at http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.22725/full.
1 Futamura, Koji, et al. Novel full-spectral flow cytometry with multiple spectrally-adjacent fluorescent proteins and fluorochromes and visualization of in vivo cellular movement. Cytometry Part A 2015:87:9 pp. 830-842 doi:10.1002/cyto.a.22725 http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.22725/full
2 Tomura M, Hata A, Matsuoka S, et al. Tracking and quantification of dendritic cell migration and antigen trafficking between the skin and lymph nodes. Scientific Reports. 2014;4:6030. doi:10.1038/srep06030. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4129424/