Zona-free oocytes were incubated in mKSOM with 1μM fura-2 AM/0.02% pluronic F-127 (Invitrogen, Carlsbad, CA) prior to addition of 10μM PF04594755 or 0.1% DMSO as a control. After a 15 minute fertilization period, the fertilization droplets were monitored at 37°C on a Nikon TE2000-U microscope equipped with a Lambda 10-2 Optical Filter Changer (Sutter Instruments, Novato, CA, USA). Fura-2 was excited at 340 and 380 nm and emissions at 530 nm were collected at 15 second intervals for 45 minutes. The fluorescence excitation ratio at 340/380nm was monitored and quantified by Metafluor software (Meta Imaging Devices, Downington, PA, USA).
Lambda 10 2 optical filter changer
Manufactured by Sutter Instruments
Sourced in United States, Germany
The Lambda 10-2 Optical Filter Changer is a laboratory equipment device designed to automatically control the positioning of optical filters. It allows for the rapid and precise selection of various filters for use in optical experiments or imaging applications.
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Lab products found in correlation
Axioskop 2 plus, by Zeiss (2 mentions)
Fura 2 am, by Thermo Fisher Scientific (1 mentions)
Pluronic f 127, by Thermo Fisher Scientific (1 mentions)
Orca flash 4.0 lt scmos camera, by Hamamatsu Photonics (1 mentions)
Mitosox probe, by Thermo Fisher Scientific (1 mentions)
Mitotracker green fm, by Thermo Fisher Scientific (1 mentions)
2 protocols using lambda 10 2 optical filter changer
1
Monitoring Fertilization Dynamics in Zona-Free Oocytes
2
Mitochondrial Membrane Potential and Oxidative Stress
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ΔΨm and mtROS were determined in neuron cultures by staining with 100 nM TMRM and 2.5 μM MitoSOX probes (Invitrogen), respectively [12 (link)]. In the TMRM staining, the non-potentiometric probe MitoTracker Green FM (100 nM, Invitrogen) was also included to normalize TMRM signal by the signal of mitochondrial mass. Images were acquired in an Axioskop2 Plus (Zeiss, Oberkochen, Germany) with Lambda 10–2 Optical Filter Changer (Sutter Instrument, Novato, California, USA) coupled to an ORCA-Flash4.0 LT sCMOS camera (Hamamatsu, Hamamatsu, Japan) from Optical and Confocal Microscopy Facility at CBMSO. Specificity of TMRM staining was assessed by the addition of 5 μM oligomycin or 5 μM FCCP to collapse ΔΨm.
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