Journal: Methods in enzymology

Article Title: Quantifying Size and Number of Adipocytes in Adipose Tissue

doi: 10.1016/B978-0-12-411619-1.00006-9

Figure Lengend Snippet: Adipocytes size and average area are similar when quantified by MetaMorph or ImageJ. Ten-week-old C57BL6/J mice (N=5) fed a 60% high-fat diet for 6 weeks were sacrificed and epididymal adipose tissue depots harvested, fixed, sectioned and adipocytes counted using either the MetaMorph or ImageJ method described herein. The frequency distribution of adipocyte sizes is equivalent whether analyzed by MetaMorph or ImageJ. No significant differences were found in either the frequency distribution (A, two-way ANOVA with Bonferroni post-hoc analysis) or average adipocyte area (B, Student’s t-test).

Article Snippet: A semiautomated custom image analysis protocol was developed using MetaMorph image analysis software to quantify the area of the individual adipocytes.

Techniques:

Journal: Methods in enzymology

Article Title: Quantifying Size and Number of Adipocytes in Adipose Tissue

doi: 10.1016/B978-0-12-411619-1.00006-9

Figure Lengend Snippet: Inguinal adipose tissue has smaller adipocytes than epididymal adipose tissue. Ten-week-old C57BL6/J mice (N=5) fed a 60% high-fat diet for 6 weeks were sacrificed and epididymal (eWAT) and inguinal (iWAT) adipose tissue depots harvested, fixed, sectioned and adipocytes counted using the MetaMorph method described herein. The distribution of adipocyte areas indicates iWAT has greater numbers of adipocytes with areas below 2000 μm2 and fewer adipocytes with areas greater than 2000 μm2 when compared to eWAT (A, B). The area-under-the-curve of this distribution quantifies this elevated number of smaller (≤2000 μm2) and decreased number of larger (≤2000 μm2) adipocyte in iWAT. The difference in adipocyte size is evident in representative H&E stained samples of both iWAT and eWAT (C). Two-way ANOVA with Bonferroni post-hoc analysis, *p<0.05 compared to respective adipocyte area increment (A and B).

Article Snippet: A semiautomated custom image analysis protocol was developed using MetaMorph image analysis software to quantify the area of the individual adipocytes.

Techniques: Staining

Journal: Methods in enzymology

Article Title: Quantifying Size and Number of Adipocytes in Adipose Tissue

doi: 10.1016/B978-0-12-411619-1.00006-9

Figure Lengend Snippet: The frequency of small adipocytes but not the average adipocyte area changes in epididymal adipose tissue located on the testicle. Ten-week-old C57BL6/J mice fed a 60% high-fat diet for 6 weeks were sacrificed and epididymal WAT harvested and divided into five equal sections according to the proximity to the testicle before fixing, sectioning and counting of adipocytes (~1000 per sample) using the MetaMorph method described herein. Whereas the average size of adipocytes did not differ between samples (A), there was a higher frequency of small adipocytes (1000–2000 μm2) directly adjacent to the testicle (labeled 1). Two-way ANOVA with Bonferroni post-hoc analysis, *p<0.05 compared to samples 2–5.

Article Snippet: A semiautomated custom image analysis protocol was developed using MetaMorph image analysis software to quantify the area of the individual adipocytes.

Techniques: Labeling