Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Aug 26;11:4275. doi: 10.1038/s41467-020-18046-y

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 3 — Optimal results of experiments can be obtained by following these steps: 1. Experimental design. 2. Tissue processing for optimal histological analysis. 3. Sectioning into thick (e.g., 40 μm) sequential sections and store in long-term tissue storage reagent (e.g. cryoprotectant). 4. Selection of representative sections using systematic random sampling (SRS). 5. Histological staining of the tissue with appropriate antibodies for cell types of interest. 6. Establishing the height of counting frame for optical fractionator (h). 7. Establishing the area sampling fraction (asf) to define the fraction of the specimen that will be quantified to represent the entire specimen. This is done by setting the sampling grid and defining the size of the counting frame (X and Y dimensions) for optical fractionator. 8. Quantification of sampled regions using optical fractionator and calculation of total numbers of markers in the specimen using the parameters set above and the number of cells counted.