Ix73 epi fluorescent microscope

Fluorescence and DIC images were acquired on an Olympus IX73 epi-fluorescent microscope fitted with an Andor iXon Ultra 888 EMCCD and controlled by MetaMorph (v7.8). Fluorescence illumination was provided by a Lumencor Spectra X light engine containing a solid state light source. DAPI: excitation filter = 390 (40) nm, emission filter = 435 (48) nm. GFP: excitation filter = 482 (18) nm, emission filter = 528 (38) nm. NIR: excitation filter = 640 (14) nm. Emission filter = 705 (72) nm.

HeLa Kyoto cancer cells were seeded on to an eight well chamber slide (Ibidi) at a density of 1 × 10⁴ cells per well 24 h before imaging. Cells were cultured in Dulbeccos Modified Eagles Media supplemented (DMEM) with 10% fetal bovine serum (FBS), 1% L-glutamine, and penicillin-streptomycin (1000 U/mL), and incubated at 37 °C and 5% CO₂. The slide was place on the microscope stage surrounded by an incubator to maintain the temperature at 37 °C and CO₂ at 5%. J-aggregated 1 (8 µM) was added to cells and images acquired after 1 h incubation. DIC imaging was used to choose a field of view and focus on a group of cells. Fluorescence and DIC images were acquired on an Olympus IX73 epi-fluorescent microscope fitted with an Andor iXon Ultra 888 EMCCD and controlled by MetaMorph (v7.8). Fluorescence illumination was provided by a Lumencor Spectra X light engine containing a solid-state light source. NIR: excitation filter = 640 (14) nm, emission filter = 705 (72) nm. Images were acquired using a 60×/1.42 oil PlanApo objective (Olympus). Image processing was completed by using software ImageJ 1.52n (National Institutes of Health, USA). SRRF imaging were obtained from a stream of 100 images taken at frame rates of 35 - 50 per sec with Image J plugin Nano-J-SRRF used to generate high quality SRRF images.

MDA-MB 231 cells obtained from Caliper Life Sciences. MDA-MB 231 human breast cancer cells were seeded on to an eight well chamber slide (Ibidi) at a density of 1 × 10⁴ cells per well 24 h before imaging. Cells were cultured in Dulbeccos Modified Eagles Media supplemented (DMEM) with 10% fetal bovine serum (FBS), 1% L-Glutamine, and penicillin-streptomycin (1000 U/mL), and incubated at 37 °C and 5% CO₂. The slide was place on the microscope stage surrounded by an incubator to maintain the temperature at 37 °C and CO₂ at 5%. DIC imaging was used to choose a field of view and focus on a group of cells. Fluorescence and DIC images were acquired on an Olympus IX73 epi-fluorescent microscope fitted with an Andor iXon Ultra 888 EMCCD and controlled by MetaMorph (v7.8). Fluorescence illumination was provided by a Lumencor Spectra X light engine containing a solid-state light source. NIR: excitation filter = 640 (14) nm, emission filter = 705 (72) nm. Images were acquired using a 60×/1.42 oil PlanApo objective (Olympus). Image processing was completed by using software ImageJ 1.52n (National Institutes of Health, USA).