Cfi60 plan apochromat lambda 40 objectives

Study 1: a Nikon NIU microscope equipped with CFI60 Plan Apochromat Lambda 10,40, 60, and 100× Objectives, an Andor Clara Digital Camera, and a 64‐bit imaging workstation running NIS Elements software (Nikon, U.S.A). Study 2: Nikon Eclipse NI‐E microscopes equipped with CFI60 Plan Apochromat Lambda 40× Objectives, C‐FL AT GFP/FITC Long Pass Filter Sets, Hamamatsu ORCA‐Flash 4.0 cameras, and 64‐bit imaging workstations running NIS Elements software.

Imaging was performed on Nikon Eclipse NI‐E microscopes equipped with CFI60 Plan Apochromat Lambda 40× Objectives; C‐FL AT GFP/FITC Long‐Pass Filter Sets; Hamamatsu ORCA‐Flash 4.0 cameras; H101F—ProScan III Open Frame Upright Motorized H101F Flat Top Microscope Stages; and 64‐bit imaging workstations running NIS Elements software (Nikon; Melville, NY). For the reliability studies, these systems were programmed to automatically capture sets of 15 × 15 stitched images containing up to 5,000 sperm. For the acrosome‐exocytosis studies, the C‐FL AT GFP/FITC Long‐Pass Filter Set was modified to have a band‐pass emission filter (ET535/50M). A C‐FL At Cy5/Fluor 647/Draq5 Filter set in combination with a ND16 filter were employed to visualize the Alexa Fluor 647.