A1 resonant scanning confocal microscope a1r sime

iSLK-ORF20_STOP or TREX-BCBL1 cells were grown on coverslips and fixed in 4% formaldehyde for 20 min at room temperature. Cells were then permeabilized in 1% Triton X-100 and 0.1% sodium citrate in phosphate-buffered saline (PBS) for 10 min, saturated in bovine serum albumin (BSA) for 30 min, and incubated with the designated antibodies at a 1:100 dilution. After 1 h, coverslips were washed in PBS and incubated with Alexa Fluor 594 or Alexa Fluor 488 secondary antibodies at 1:1,500 (Invitrogen). Coverslips were washed again in PBS and mounted in DAPI-containing Vectashield mounting medium (Vector Labs) to stain cell nuclei before visualization by confocal microscopy on a Nikon A1 resonant scanning confocal microscope (A1R-SIMe). The microscopy data were gathered in the Light Microscopy Facility and Nikon Center of Excellence at the Institute for Applied Life Sciences, UMass Amherst, with support from the Massachusetts Life Sciences Center.

iSLK-ORF20_STOP or TREX-BCBL1 cells were grown on coverslips and fixed in 4% formaldehyde for 20 min at room temperature. Cells were then permeabilized in 1% Triton X-100 and 0.1% sodium citrate in phosphate-buffered saline (PBS) for 10 min, saturated in bovine serum albumin (BSA) for 30 min, and incubated with the designated antibodies at a 1:100 dilution. After 1 h, coverslips were washed in PBS and incubated with Alexa Fluor 594 or Alexa Fluor 488 secondary antibodies at 1:1,500 (Invitrogen). Coverslips were washed again in PBS and mounted in DAPI-containing Vectashield mounting medium (Vector Labs) to stain cell nuclei before visualization by confocal microscopy on a Nikon A1 resonant scanning confocal microscope (A1R-SIMe). The microscopy data were gathered in the Light Microscopy Facility and Nikon Center of Excellence at the Institute for Applied Life Sciences, UMass Amherst, with support from the Massachusetts Life Sciences Center.