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Journal of Histochemistry and Cytochemistry
Volume 52 (3): 347-354, 2004
Copyright ©The Histochemical Society, Inc.

Immunofluorescence and Confocal Laser Scanning Microscopy of Chronic Myeloproliferative Disorders on Archival Formaldehyde-fixed Bone Marrow

Rosmarie Suetterlin, Werner Baschong and R. Hubert Laeng

Maurice E. Mueller Institute at Biozentrum, Basle, Switzerland (RS,WB), and Department of Pathology, Kantonsspital, Aarau, Switzerland (RHL)

Correspondence to: R. Hubert Laeng, MD, Dept. of Pathology, Kantonsspital, CH-5001 Aarau, Switzerland. E-mail: laeng{at}ksa.ch

Spatial analysis of the histoarchitecture and photographic documentation at high resolution are the principal advantages of confocal laser scanning microscopy (CLSM) over conventional fluorescence microscopy (CFM) if combined with appropriate software. Restrictions for the use of CFM and CLSM, on the other hand, include nonspecific background fluorescence, fading of photolabile fluorochromes, and both tissue-specific and fixation-induced autofluorescence. Most of those shortcomings can now be avoided. Autofluorescence, the most limiting factor of high-resolution CLSM, was recently controlled also for paraffin sections of archival formaldehyde-fixed tissues. This allowed the present study on cytoskeletal fibers and extracellular matrix proteins in both neoplastic cells of myeloproliferative disorders and in medullary stromal cells using CLSM under proper autofluorescence control. By multiple fluorescence labeling, we found that the intracellular smooth muscle {alpha}-actin (SMA) fibers and the two extracellular adhesive matrix proteins tenascin and fibronectin vary in their presence in stromal and/or myeloid cells according to the degree of bone marrow fibrosis in chronic myeloproliferative disorders (CMPDs). CLSM offers further insight in our attempts to understand a complex interplay between the two cellular compartments. (J Histochem Cytochem 52:347–354, 2004)

Key Words: immunofluorescence • confocal laser scanning • microscopy • chronic myeloproliferative • disorders • bone marrow • fibrosis • formaldehyde-fixed paraffin • sections • archival tissue


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