Single cell sorting was performed using a MoFlo Astrios cell-sorter (Beckman Coulter Life Sciences, Indianapolis, IN, USA) as described previously [18 (link)]. Briefly, the cells were stained with DAPI (Dojindo, Kumamoto, Japan), and DAPI and AcGFP1 were excited with 355 and 488 nm lasers, respectively. In total, 1000 wild-type Pv11 cells were seeded as a feeder layer in each well of a 96-well plate prior to sorting. The sorted and the feeder cells were grown for two weeks, and then a second sorting (bulk cell sorting) was performed to eliminate the feeder cells.
Miyata Y., Tokumoto S., Arai T., Shaikhutdinov N., Deviatiiarov R., Fuse H., Gogoleva N., Garushyants S., Cherkasov A., Ryabova A., Gazizova G., Cornette R., Shagimardanova E., Gusev O, & Kikawada T. (2022). Identification of Genomic Safe Harbors in the Anhydrobiotic Cell Line, Pv11. Genes, 13(3), 406.
Other organizations :
Institute of Agrobiological Sciences, Tokyo Medical and Dental University, National Agriculture and Food Research Organization, The University of Tokyo, Skolkovo Institute of Science and Technology, Kazan Federal University, Endocrinology Research Center, Juntendo University, RIKEN Center for Integrative Medical Sciences
Number of wild-type Pv11 cells sorted and grown for two weeks
Number of feeder cells eliminated after second sorting
control variables
Staining of cells with DAPI
Excitation of DAPI and AcGFP1 with 355 and 488 nm lasers, respectively
Seeding of 1000 wild-type Pv11 cells as a feeder layer in each well of a 96-well plate prior to sorting
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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