Frozen-section fluorescence microscopy and stereology in the quanti cation of neuronal death within dorsal root ganglia
2004 (English)In: Journal of Molecular Histology, ISSN 1567-2379, E-ISSN 1567-2387, Vol. 35, no 6, 565-580 p.Article in journal (Refereed) PublishedText
Histochemical and morphological research increasingly relies upon quanti cation of complex biological systems. For such investigations to be meaningful, quanti cation techniques must meet the seemingly conflicting requirements of being theoretically robust, yet sufficiently practical to facilitate widespread applicability. Validity ought to be enhanced by theoretical simplicity, use of measured rather than assumed variables, and minimising observer interpretation. Practicality is facilitated by simplifying and reducing measurements, broadening applicability, and reducing costs and analysis time. As a result, quanti cation systems that rely upon sampling and estimation have been favoured over serial reconstruction techniques. To provide reliable estimates, sampling must be valid at all levels from tissue harvest, to the selection of microscope fields in which quanti cation is performed by techniques that account for the anisotropic distribution, and variable size of many elements in biological systems. These principles are embodied in the development of a stereological approach to the quanti cation of neuronal death within dorsal root ganglia after peripheral nerve injury. This frozen section technique is efficient and flexible, since it permits simultaneous morphological examination, TUNEL, or standard fluorescence immunohistochemistry, broadening its applicability. Section shrinkage is minimal, and counting by optical disection has proved to be time-efficient and sufficiently reproducible to reliably detect losses in the order of 5%, with minimal inter-observer variation. As is discussed, stereology has not yet met with universal acceptance, but by balancing theoretical validity with practical applicability, it has proved an excellent approach to the investigation of neuronal death within dorsal root ganglia.
Place, publisher, year, edition, pages
Dordrecht: Kluwer Academic Publishers, 2004. Vol. 35, no 6, 565-580 p.
primary sensory neurons, counting methods, cell-death, pathological research, unbiased estimation, peripheral axotomy, tissue-sections, disector method, sciatic nerve, time-course
Cell and Molecular Biology
IdentifiersURN: urn:nbn:se:umu:diva-122161DOI: 10.1007/s10735-004-2187-5ISI: 000225903600004PubMedID: 15614610OAI: oai:DiVA.org:umu-122161DiVA: diva2:941253