This is a basic difference to film exposures with their limited density range of and a fixed exposure time resulting from the film sensitivity (ISO film speed) of the selected film system class and the density requirements.
Briefly, fluorescence images of the posterior most coelomocytes (either ccDL or ccDR) were captured using a fixed exposure time of 360 milliseconds.
A Hamamatsu CCD camera and Openlab software (Improvision) were used for image acquisition at the brightest focal plane and a fixed exposure time.
Images were collected at fixed exposure times using a filter set optimized for dityrosine fluorescence (Ex. 320 nm; Em 410 nm) (Omega Optical, Brattleboro, VT).
Images were obtained with a Zeiss Axioplan 2 microscope with Axiovison software (fixed exposure time 728 ms), and immunofluorescence was quantified in a set of regions of interest (14300 pixels), using Image J 1.40 from NIH, Bethesda, Maryland, USA (http://rsb.info.nih.gov/ij/).
Images were captured using an LSM 510 confocal microscope (Zeiss) using a fixed exposure time.
All images were acquired with the MetaMorph software using a fixed exposure time and gain.
For analysis of superoxide levels, 5 8 images of each heart sample were captured using fixed exposure time for all groups.
Coverslips with adhering cells were photographed in randomly selected fields under a ×40 objective while maintaining fixed exposure parameters.
