The evaluation of a special liquid fixative for direct immunofluorescence

Skeete, M.V.H.; Black, Martin M.

doi:10.1111/j.1365-2230.1977.tb01537.x

Cited by 32 publications

(9 citation statements)

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“…If biopsies are kept longer than 24 hours in saline, decreased fluorescence of tissue-bound immunoreactants may be encountered, although we have observed bright specific fluorescence of the BMZ in biopsies kept in saline for at least 5 days. Michel's fixative seems to have a similar limitation: Skeete and Black found that biopsies stored in this fixative should be received within 1 day of biopsy for consistently reliable results [4]. …”

Section: Discussionmentioning

confidence: 99%

Enhanced diagnostic immunofluorescence using biopsies transported in saline

et al. 2004

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BackgroundThe demonstration of tissue-bound immunoreactants by direct immunofluorescence microscopy (DIF) is a valuable parameter in the diagnosis of various autoimmune and immunecomplex-mediated skin diseases. For preservation of tissue-bound immunoreactants, biopsies are usually fresh-frozen in liquid nitrogen or transported in Michel's fixative. But even optimally preserved tissue specimens are no guarantee for the correct diagnosis by DIF, especially when weak to moderate IgG fluorescence of the epidermal basement membrane zone is involved. In such cases false negative results are easily obtained due to the relatively high dermal "background" fluorescence produced by polyclonal anti-human IgG fluorescein conjugates.MethodsIn the present study we have compared the use of normal saline (0.9% NaCl) with liquid nitrogen and Michel's fixative as transport medium for skin biopsies. From 25 patients with an autoimmune skin disease (pemphigus, pemphigoid, lupus erythematosus and vasculitis) four matched skin biopsies were obtained and transported in either saline for 24 and 48 hours, liquid nitrogen, or Michel's fixative for 48 hours.ResultsDirect IF microscopy showed significant reduction of background fluorescence (p < 0.01) and relatively enhanced desired specific (IgG, IgA) staining in biopsies transported in saline. A conclusive or tentative IF diagnosis was reached in 92% after 24 h saline, 83% after 48 h saline, 68% after freezing in liquid nitrogen, and 62% after 48 h Michel's medium (n = 25).ConclusionsWe conclude that transporting biopsies without freezing in normal saline for 24 hours is an adequate and attractive method for routine IF diagnosis in autoimmune and immune complex-mediated dermatoses. The superior results with saline incubation are explained by washing away of IgG background in dermis and epidermis.

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Section: Discussionmentioning

confidence: 99%

Enhanced diagnostic immunofluorescence using biopsies transported in saline

et al. 2004

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“…A previous study by Skeete and Black [3] suggested that after long intervals in Michel's medium the intensity of fluorescence might become absent in some cases where snap-frozen sec tions were positive. This tends to occur because catabolism of blood and serum proteins produces a drop in the pH of the medium.…”

Section: Discussionmentioning

confidence: 99%

A Retrospective Analysis of Tissue-Fixed Immunoreactants from Skin Biopsies Maintained in Michel’s Medium

et al. 1994

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656 skin biopsies with positive direct immunofluorescence from the UK and overseas were studied over a 2-year period. The length of time biopsies had remained in Michel’s medium at pH 7.0 in various diseases (pemphigoid, pemphigus, linear IgA disease, epidermolysis bullosa acquisita, lupus erythematosus, vasculitis, amyloid, lichen planus and dermatitis herpetiformis) was analysed. We concluded that direct immunofluorescence remained positive at 6 months and that Michel’s medium is a reliable long-term maintenance medium for skin biopsies.

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“…However, it was claimed that sections obtained for routine histology were not always as good as those after formalin fixation. Recently, in a comprehensive study, Skeete and Black (1977) investigated the value of the method described by Michel et al (1973). It was concluded that the liquid fixative was useful but consistently reliable only if the specimens were received within one day of biopsy.…”

Section: Discussionmentioning

confidence: 99%