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Detailed step-by-step procedures for microscope setup were published earlier (Gonen, 2013 ▸).
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For the F/G actin quantifications, the same microscope setup was used.
Our microscope setup was able to provide linear temperature gradients within an acceptable range.
The wide-field epifluorescence microscope setup was controlled by Metamorph 7.5 (Molecular Devices, Sunnyvale, CA).
The performance of our microscope setup was tested during ESA's 53rd PFC operated by Novespace in October 2010.
Thus, optimized microscope setup is required to avoid spectral overlap between Nile red and GFP fluorescence emission.
Provided the microscope setup is equipped with the appropriate filter sets, first superficial information on the intracellular distribution can be obtained from conventional fluorescence microscopy.
For AF studies, microscope excitation setup was adjusted to obtain a light fluence rate quite comparable to that used for microspectrofluorometric analysis (6.35 mJ cm−2 per second).
A previously described confocal microscope setup (29) was used to image immunostained islets with excitation/emission at 488/527 nm for Alexa Fluor 488 and 561 and/or >645 nm for RFP.
The optical setup is a coherent microscope based on DH in Mach-Zehnder configuration interferometer as illustrated in Fig. 1.
The setup was visualized by a microscope (at 600-fold magnification) connected to a digital camera (Axis, Lund, Sweden).
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