Uplans apo 100x objective lens

Immunofluorescence images were taken using an Olympus UPlanS APO 100X objective lens (numerical aperture 1.40) on an Olympus IX-70 microscope equipped with the DeltaVision RT (Applied Precision) imaging system. Z-section image series were collected at 0.2 μM intervals over a total of 3–4 μM through the center of the cells and deconvolved using softWoRx (Applied Precision).

Images were taken using an Olympus UPlanS APO 100X objective lens (numerical aperture 1.40) on an Olympus IX-70 microscope equipped with the DeltaVision RT (Applied Precision, GE Healthcare) imaging system. Z-section image series were collected at 0.2 μM intervals over a total of 3–4 μM through the center of the cells. All protein and DNA localization was done with images that had been deconvolved using SoftWoRx (Applied Precision, GE Healthcare).
The images were converted into RGB.tif files using the image processing software Fiji. To reduce background, the red, green and blue channel levels were all set to a minimum level of 25 on a scale from 1 to 255 using Photoshop.

Cells were imaged on a DeltaVision wide-field fluorescence microscope (Applied Precision, Inc.). It has a digital camera (CoolSNAP HQ2; Photometrics). An oil immersion Olympus Plan Apo 60x objective lens (N.A.=1.42), Olympus UplanSApo 100x objective lens (N.A. = 1.40), or Olympus LUCPlanFLN 20x objective lens (N.A. = 0.45) was coated with 1.518 refraction index low autofluorescence immersion oil, Olympus Type F (Fischer Scientific, NC0297589). A 250 watt Xenon Arc lamp was used to direct excitation lighting from the back of the microscope and focused from below onto the coverslip held on an Olympus IX-71 stage. Dichroic filter set uses the Alexa setting: FITC excitation: 475/28 Emission: 523/36; A594 excitation: 575/25 emission: 632/30; CY5 excitation: 632/20 emission: 67634; DAPI excitation: 390/18 emission: 435/38. Exposure times varied depending on the type of experiment and staining conditions.