Here, we seek to provide a thorough assessment of such a PIV-based approach through synthetic datasets, in which PIV parameters, such as the camera acquisition rate and the size of the interrogation area, are systematically varied, without experimental confounds.
The raw camera acquisition rate was 25 frames/s (40 ms per frame).
Then, on the basis of radial metrology concepts, each single frame of the loading sequence is automatically processed to track 3-D marker positions and, thus, to obtain sample deformation at the camera frame acquisition rate.
The microscope used in this study was a Zeiss Axiovert 200 M inverted microscope (75 W Xe lamp, Zeiss, Germany) fitted with a 100×/1.3 NA oil-immersion microscope objective and an Andor iXon3 EMCCD camera (20 fps acquisition rate).
An optical motion tracking system (VICON, Oxford, UK) consisting of nine high speed MX-13+ cameras was used at acquisition rate of 150 Hz.
Emitted light was filtered (530 560 nm band-pass) and imaged using a high sensitivity, electron multiplying CCD camera (iXon, Andor Technology) at an acquisition rate of 20 frames per second.
To accomplish the high-speed imaging and collection, the CMOS camera (Mintron, MC1310) has an acquisition rate of 500 frames/s at the full resolution of 1000 (H ) × 512 (V ) pixels, with square pixels of 12 μm.
While this is a good level of sensitivity it can be further improved as discussed in the conclusion, it is clear that, apart from the acquisition rate, the camera system does not impose a limit of the achievable performance.
However, wide-field detection requires the use of cameras, which limit the data acquisition rate to 1 kilocycle per second or slower using state-of-the-art charge couple device (CCD) cameras with on-chip amplification.
The image acquisition rate of each camera was equal to 2 Hz.
The presented method calculated automatically the position of the eye centre (x- and y-axis) and both upper and lower beak tips distance in a high level of accuracy, but the model can be improved by using a camera with higher resolution, a higher acquisition rate, and infrared-reflective markers.
