How to count multilocular adipocytes on a slide with a white adipose tissue sample stained with H&E? I had done the count on the entire slide, but the articles put it in % of cells or adipocytes/mm². The total area of the slide is too large to be photographed under the microscope at 5X. And I can only be sure if it is multilocular with the 40x lens. So, I have no way of taking photographs and measuring them later in the ImageJ program. The articles do not detail how I do this measurement. The objective is to compare whether one group has more multilocular adipocytes than another: checking browning activity. Please if you can help, I would appreciate it
I would recommend scanning the whole slide and creating a digital image and then using QuPath (https://qupath.github.io/), which is open source and free, to anotate and analyse the areas.
Hi Marilia Cavalcanti,
If you already counted the total adipocytes in the sample, you will certainly need to measure the sample area. Without a scanner, you can try making a collage of photomicrographs of all the sample, but it can be tricky.
Another way: I think you could estimate adipocyte/mm² by counting adipocytes in a certain amount of photomicrographs of your sample taken with the 40x lens. Then, calculate the area of a photomicrograph if you know the calibration and do the calculations to obtain how many adipocyte are per mm² of the sample.
For example: Take 10 photomicrographs of sample 1 with 40x lens. Count white and brown adipocytes (WA, BA) in each image using ImageJ/FIJI (you can use Cell Counter plugin) or any other software. Estimate the total area of the image at 40x. Obtain the WA/area and BA/area of each image and calculate the mean for sample 1. Then, do the math to obtain WA//mm² and BA/mm².
Hope it helps.
Good luck
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