Stacking and Stitching of photomicrography
Microscopes are excellent tools for revealing the great details of tiny things but the pictures taken from the microscope are limited to the size of the field of view and furthermore limited by the size of your camera sensor. If you want to take picture of a subject larger than the field of view, you would need to take series of photos containing part of the subject then stitch them together. It's further complicated by the dept of focus. You would need to stack series of photos focusing on different focal plane. Here, I am demonstrated the use of freely available software allows you to stacking and stitching a silverfish photos taken from SM-Plus stereo microscope.
1. Take photos for each part of the silverfish for each focal plane. You can either take photos from upward or downward on the focal plane. I am going from downward in one area then move to the next area in upward direction.
Figure 1. Individual photos of silverfish. Each photo represents one area of the silverfish on each photo plain.
Figure 2. The above picture is one of the component picture from above. The focus plane was right around the edge of the insect and the eyes. The tips of the antenna and top of the thoracic are fuzzy.
2. Stacking the photos with Combine ZP. This step will produce the stacked photos of each area. The software is extremely easy to use. Just select the desirable stacking method than click on GO. The stacking involved in several processes, such as balance color and brightness, frame alignment, finding details, noise removal, gap filling and filters. If you like to tweak your own stacking method or experience the stacking process, you can enable the menu. The software can be downloaded from http://hadleyweb.pwp.blueyonder.co.uk/CZP/Installation.htm.
Figure 3. Combine ZP photo stacking software package
3. Once I completed each part of the silverfish, I began to assemble the stacked photos into one single picture with Microsoft Image Composite Editor. As you can see, the first pass was not very successful. The three spikes at the end of abdomen were lost during the assemble process. My solution was to assemble the terminal segment of the abdomen and three spikes first then assembled the whole insect.
Figure 4. Microsoft Image Composite Editor - Panorama
Figure 5. Composite the three spikes and the end of the abdomen first
Figure 6. Composite the photos in two passes, first with the end of abdomen then the complete picture, I successfully assemble the whole insect.
4. Painted over the edges of the background with gray. Ideally, the photo of the background should also be taken to prevent the need to paint the background.
Figure 7. Completed photos of the silverfish with photo stacking and stitching (Due to the limitation of the content management system, I have to reduce the file size from 7010 x 17804 to 631 x 1600 to allow file upload.)
For compairson, another photo was taken with Olympus Stylus in super macro mode. Although I can get close enough and take the shot of the whole insect, the macro shot lacks much details of the insect.
Figure 8. Macroshot taken with Olympus Stylus 7010.
If you would like to practice the stacking and stiching the silverfish, I made some smaller files that you can down load and try it yourself. It can be download here:
http://www.onfocuslaboratories.com/support/files/silverfish_small_version.zip
