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Cfi plan fluor 40x

Manufactured by Nikon

The CFI Plan Fluor 40X is a microscope objective lens designed by Nikon. It provides a 40X magnification and is suitable for fluorescence microscopy applications.

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2 protocols using cfi plan fluor 40x

1

Multiplexed Protein Interaction Analysis

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Images were captured by inverted epifluorescence microscope (NIKON TI-E) equipped with Photometrics CoolSNAP HQ2 CCD camera and an automatized stage for both chip (CFI Plan Apo VC 60XWI, NIKON) and glass slide (CFI Plan Fluor 40X). Image acquisition was performed by using NIKON software. Z-stack images were first treated with Fiji, using background subtraction, gamma and maximum intensity projection functions. Then, PLA signals were quantified using CellProfiler (2.2.0, rev ac0529e) in the following modules: Identify primary objects/identify objects manually for defining nuclei using DAPI staining; identify secondary object for defining cytoplasm using cytokeratin (CK) staining or DAPI staining with N-distance method; enhance speckles features to enhance PLA signals and then identify primary objects for defining PLA signals. For each cell, the presence of cytokeratin staining was determined using the thresholding based on fluorescence signal intensity on the background. Object enhancing and filtering size were adjusted depending on the objective used for control experiments on chip and on glass slides, which also varied between patient samples.
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2

Optical Manipulation and Imaging Setup

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A sketch of the experimental setup is shown in Figure 4. A 660 nm laser beam (Laser Quantum, Opus 660) was expanded with a 5× beam expander (Thorlabs, GBE05-A) and directed to a Nikon inverted microscope (Nikon, Ti-E) with a 100× oil objective (Nikon, CFI Plan Fluor 100XS Oil) for the manipulation experiments inside a microfluidic chamber of ∼120 μm thickness. A Nikon upright microscope (Nikon, Eclipse Ni) with 40× air objective (Nikon, CFI Plan Fluor 40X) was used for automated manipulation experiments. For the dark-field optical imaging, an air condenser (Nikon, C-AA Achromat/Aplanat Condenser) was used to focus the incident white light onto the sample from the top. A complementary metal-oxide-semiconductor (CMOS) camera (Nikon, DS-Fi3) or a charge-coupled device (CCD) camera (Lumenera, 12 fps) was used to record the optical images. A Notch filter (658 nm) was placed between the objective and camera to block the incident laser beam. White light was directed from the top (bottom for the upright microscope) of the stage for bright-field imaging. A xenon lamp (Sutter Instrument Lambda, LB-LS/30) was applied through the objective with a GFP filter cube (457–487/502–538 nm for excitation/emission) for fluorescence imaging. The notch filter was removed in fluorescence imaging. The dichroic beam splitter was selected according to the excitation, emission, and laser wavelength.
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