Image acquisition.

1. Take out a frame.

2. For GLP reasons it is advised to label the frames with an identifier. Preferentially the ID should be used, which corresponds to the ID-System used by the software. The ID pattern is the following “AAAAAA_BB_CCD_EE”, whereas “_“ is a mandatory separator.

   2.1. AAAAAA:  ID of the study in six characters,
   2.2. BB: ID of the hive in two numeric characters, e.g. “05”,
   2.3. CC: ID of the frame in two numeric characters, e.g. “03”,
   2.4. D: ID of the side of the frame e.g. “a” or “b”,
   2.5. EE: ID of the BFD, e.g. 00 for BFD 0 (the day of study start).
   An example of a label on the frame would look like: “Study1_01_02a” for a permanent label or “Study1_01_02a_00”
   for a label made specifically for the day of image acquisition.

3. For unequivocal identification of the image, the label is attached on the front side of the  frame and must be visible and to be photographed at every recording.

4. Make a picture of the frames using a fixed distance.

   4.1. The minimal photographic distance is calculated in order to allow visibility of 
         at least 75% of the bottom of a cell at the outermost rim of the image. The 
         calculation is based on an average cell diameter of 5.3mm and an average cell
         depth of 11mm.The photographic distance fulfilling of the above requirement   
         is 11/(5.3 * 0.25)/2 = 4.15 fold the long axis of the frame.
   4.2. The camera to be used should be connected to and controlled by a computer.
          The control software is necessary for a number of reasons: It enables
          triggering of the camera without the need to touch it. It enables a magnified
          live-view of the image allowing the directed focusing on the eggs. The
          camera’s autofocus will always focus on the upper rim of the cell wall.
   4.3. Ideally, a setup should be created, allowing keeping the fixed distance,
          defined illumination and minimal vibrations.
   4.4. Illumination should be optimized to minimize reflections.

5. After the pictures are made, download the pictures from the camera to the computer.
   5.1. The images have to be re-named according to the pattern  “AAAAAA_BB_CCD_EE.jpg”.
   5.2. The image files of the same frame should be stored in the same folder.