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SUMMARY Expression of human milk fat globule (HMFG1) in immunohistochemically stained sections ofbreast carcinomas was assessed subjectively and objectively from 82 women (age range 41-96 years) to determine its prognostic importance. No correlation was observed between the degree of staining and prognosis even when the subcellular distribution ofantigen expression was assessed. The total absence of staining with HMFG1 was possibly associated with a favourable outcome, although this did not quite achieve significance with the small numbers involved. The Quantimet 970 was used for objective semiautomated measurement of immunohistochemical reactions in paraffin wax sections and was found to produce better resolution and to eliminate subjective error.In his study of squamous carcinoma of the lip Broders was one of the earliest workers to note the association between the histological differentiation of a tumour and its prognosis.' The degree of differentiation of most tumours is now considered by many pathologists and clinicians to have some prognostic value. Bloom and Richardson applied this hypothesis to breast carcinomas and separated them into three histological grades.2 This grading method, which subjectively assesses tubular differentiation, mitotic index, and nuclear characteristics, has been shown to lack interobserver reproducibility and is only moderately predictive of prognosis for grades 1 (low) and 3 (high), with less certainty for grade 2.3 There has been some evidence to suggest that the combination of histological grade and tumour stage provides a better prognostic index.4The recent development of monoclonal antibodies directed against normal constituents of breast duct epithelium5 has raised the possibility that expression of these antigens may be related to the degree of cell differentiation and hence prognosis. Present methods of grading antigen expression in paraffin wax sections are subjective and lack precision. This problem may be overcome by image analysis which permits accurate quantitation of immunoreactivity.In this retrospective study of cases of known survival of breast carcinoma we present our findings on HMFGI quantitation by standard light microscopical examination and automated image analysis.

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SYNOPSIS This describes the sodium sulphate-Alcian Blue (SAB) method for staining amyloid in paraffin sections. Its value lies in the possibility of subsequent counterstaining and thus of revealing the structural relationships of amyloid.In the kidney the topical disposition of amyloid closely resembles the disposition of fibrin in the kidney of diabetics; this suggests that upset in vascular permeability plays a part in determining the site of the amyloid deposits. Furthermore, an aging process in amyloid can now be envisaged resembling the aging of extraluminal fibrin. Both materials proceed to a hyalin material that, staining like collagen, merits the name pseudo-collagen. This term we apply to a hyalin, staining like collagen, for which,we can postulate a specific precursor.The I ight microscopist has generally accepted amyloid as a hyalin substance, an acellular firm gel, situated interstitially, and distinguished from other hyalins by particular staining reactions, notably the metachromatic reaction with methyl violet and an affinity for Congo red. Neither of these methods is ideal for the study of the structural relationships of amyloid.Our attempts to demonstrate and study amyloid, as seen in postmortem material, were progressing unprofitably when suddenly one divagation revealed possibilities. A rationally evolved although perhaps inaccurate idea led one of us (W.S.) to produce a modified Alcian Blue solution. This enabled us to stain amyloid, in sections from paraffin, with a dye that could be stabilized in situ and thus allow a variety of counterstainings. Although, as with most other dye-staining methods, the results were not chemically specific for amyloid, with suitable counterstaining they proved sufficiently selective to allow study of the precise situation of the amyloid deposits. Unfortunately the dye makers, in 1955, changed the constitution of the dye, and our results altered miserably.