Cutting (20microns/section)

1. When you reach the muscle, cut 8 muscle sections and place each section on 1 slide. So after cutting these 8 sections you should have 8 slides with 1 muscle section each 2. Then, cut and discard 8 muscle sections 3. Follow that by again by cutting 8 sections and put 1 of each on the 8 slides you used in step 1. You should now have 8 slides with 2 muscle sections each. 4. Repeat Steps 2 and 3 until you have filled the slides (10 muscle sections per slide) 5. Once you have filled that set of slides, start another set of 8 slides and repeat steps 1-3. Keep filling slides until no more muscle is left

Counting (goal = ~125-150 macrophage per muscle)

1. You should have 10 muscle sections per slide (for the last set of slides you may only have half of the slide filled depending on how big the muscle was, so adjust counting as necessary) 2. You want to count approximately the same number of macrophages per slide with approx 125-150 total count. So if you have 3 slides with 10 muscle sections, I counted 50 macrophages per slide. When my 3rd slide only had 5-6 muscle sections on it, I would count 50 macrophages from slides 1 and 2 and 25 macrophages from slide 3 to get 125 macrophages total 3. For each slide, I would start out by counting all the macrophages in the 1st, 3rd, 5th, 7th, and 9th cut sections (also have some sections from the edge of the muscle and from the center). I arranged my sections horizontally when cutting on the cryostat, so the sections I counted were these (red sections):

10

4. If I was not able to reach 125-150 macrophages using these 5 sections, I would then also count the 2nd, 4th, 6th, 8th, and 10th sections until I reached the goal*

*For the runner muscles, sometimes it was difficult to count up to 125-150 macrophages because there seemed to be fewer macrophages. In that case, I just counted all the macrophages I could find in that slide