Cfi plan apo lambda 100

Prior to imaging, 1 mL cell culture was concentrated to 50 µL by centrifugation at 1500 x g, 30 s. 2 µL cell suspension was loaded under a 22 × 22 mm glass coverslip (VWR, thickness: 1.5).
Spinning-disk confocal images were captured with Eclipse Ti-E inverted microscope fitted with Yokogawa CSU-X1 spinning disk confocal scanning unit, equipped with ILE 405 nm 100 mW, ILE 488 nm 50 mW and ILE OBIS 561 nm 50 mW laser lines, Nikon CFI Plan Apo Lambda 100× (N.A. = 1.45) oil objective and Andor iXon Ultra U3-888-BV monochrome EMCCD camera. Imaging was performed at 30 °C with acquisition of 13 z-slices, 0.58 µm per slice, controlled by either Andor iQ 3.6.5 (Oxford Instruments) or Andor Fusion software 2.3.0.36 (Oxford Instruments). Temperature control was achieved by the Okolab cage incubator set at 30 °C.
Epifluorescence brightfield images were acquired using Zeiss Axio Observer Z1 fluorescence microscope fitted with α Plan-FLUAR 100×/1.45 NA oil objective lens (Carl Zeiss) and the Orca-Flash4.0 C11440 camera (Hamamatsu). All images were taken at the medial focal plane, controlled by Zen 2012 software (blue edition, Carl Zeiss Microscopy GmbH).

The microscope system consists of a Nikon Diaphot TE300 microscope equipped with CFI Plan Fluor 60× oil, numerical aperture (NA) 1.4 and CFI Plan Apo Lambda 100× oil, NA 1.45 objectives, a 10× projection lens (Nikon, Garden City, NY) and a Prior motorized stage. Fluorescent images were obtained with a SensiCamQE CCD camera (Cooke Corp., Romulus, MI) using dual filter wheels (Prior) with single band excitation and emission filters for FITC/TRITC/CY5/DAPI (Chroma, Rockingham, VT). Cell images were processed and analyzed using IP Lab software version 3.9.4r4 with a fluorescence colocalization module (Scanalytics, Inc., Fairfax, VA). Images acquired using the 60× objective have dimensions of 78 µm × 58 µm and images acquired using the 100× objective are 41 µm × 30 µm. Calibration bars on images are 10 µm.