When photographing macro subjects, it's important to pay attention to the angle of the digital sensor (i.e. the back of the camera) relative to the subject. If the angle is oblique, sufficient depth of field will be hard to achieve. If the sensor is parallel with the subject, you can obtain good DOF even with larger lens apertures because all points on the sensor will be approximately equidistant to the subject. To fill most of the frame with this spotted glass frog in Costa Rica, I used a Canon 100-400mm lens plus one extension tube. The tube allowed me to focus closer than the minimum focusing distance of the lens. My settings were 1/500, f/10, 8000 ISO, and then I reduced the noise with Neat Image software.
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