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Optical Disector Demonstration for Morphometric Analysis

Learn about optical disector technique with an animated cartoon, exploring counting nuclei in consecutive optical sections of rabbit testis. Understand the counting frames and rules for accurate morphometric research.

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Optical Disector Demonstration for Morphometric Analysis

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  1. Demonstration of the optical disector Zheng-Wei Yang Morphometric Research Laboratory North Sichuan Medical College Nanchong city, Sichuan province, China (This English version is written on 2 July 2010 as suggested by Nigel G. Wreford & Robert I. McLachlan, Australia)

  2. Introduction to the animated cartoon on page 4: The cartoon consists of 15 consecutive optical sections (spaced 1 m apart) observed through a high aperture oil immersion lens within a single 25 m thick methacrylate-embedded (physical) section of rabbit testis. Particles seen in the sections are nuclei of spermatids approximating spheres. Superimposed on the sections are 4 counting frames (each 21  15 m), around which the profiles (sections) of 16 nuclei (a-p) formed on the optical sections Nos. 3-13 are indicated.

  3. Question: If we count nuclei from the 3rd optical section down to the 13th optical section, i.e. count the nuclei within the 10 m (thickness) of section between the 3rd and 13th sections, which nuclei should be counted “within” the 4 counting frames (disectors)? • Answer: Starting at the 3rd optical section all nuclei are counted as they come into focus “within” the counting frame according to the disector counting rule[1-2]. That is, we count newly appearing nuclei. See page 5 for further answers. • References • Sterio DC. The unbiased estimation of number and sizes of arbitrary particles using the disector. J Microsc 1984; 134(Pt 2): 127-36. • Gundersen HJ et al. The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. APMIS 1988; 96(10): 857-81.

  4. Play this slide to see an animated cartoon showing the optical disector

  5. Upper left frame: All 3 nuclei (a, b, c) should be counted. • Upper right frame: 4 nuclei (d, e, f, g) should be counted; 1 nucleus (h) should not. • Lower left frame: 2 nuclei (i, j) should be counted; 1 nucleus (k) should not. • Lower right frame: 4 nuclei (m, n, o, p) should be counted; 1 nucleus (l) should not.

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