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The PEFC was set on an automatic rotating stage, and a picture was taken every 2.25° by using a cooled CCD camera with grey scale of 16 bit.
A xenon flash-lamp is used as the single excitation source for the four sensing regions, while multichannel phosphorescence detection is achieved by using a cooled intensified charge-coupled device (ICCD).
Digital images were obtained by using a cooled CCD camera, Coolsnap HQ (Photometrics), coupled to a Leica DMRA2 microscope using the same exposure time for all aquisitions.
Fluorescence emission was imaged by using a cooled charge-coupled device camera; the exposure times were 500 ms for CFP and YFP images.
The best capture efficiency for each section was obtained by using a cooled slide (4°C) that was fixed 30 sec with a ice cold 70% ethanol solution (-20°C).
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The other method is by using a cooling machine attached to a cap, which ensures a more constant temperature.
Losses of volatiles, through the gas exhaust port, were minimized by using a cooling condenser attached to a RTE7 water bath (Thermo Fisher-Scientific, Sunnyvale, CA, USA) kept at 4 °C.
The condenser heat removal is done either by using a cooling tower or through cold air circulation in the condenser.
This is achieved by using a cooler branding iron and applying it to the coat for a shorter time so as not to kill the hair growth cells of the branded area.
All images of each specimen were captured by using a cool CCD camera (Media Cybernetics, Silver Spring, MD, USA).
Therapeutic hypothermia was started immediately after admission to the emergency department and was applied by using a cooling technique combining the administration of intravenous ice-cold fluids and the application of a surface cooling device (Arctic Sun System; Medivance, Louisville, CO, USA), according to the protocol in use in our institution [ 6, 12].
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