LAM KID

[A simple method to detect urate crystals in formalin-fixed tissue]

BÉLY Miklós, KRUTSAY Miklós

OCTOBER 04, 2013

LAM KID - 2013;3(03)

[In our previous study we refuted the thesis that sodium urate crystals are not, or only rarely detectable in formalin-fixed histological samples because they dissolve in the aqueous formalin solution. Our observations indicate that dissolution of urate crystals is primarily caused by haematoxylineosin staining. Undeniably, however, urate crystals are partially dissolved in the aqueous solution of formaldehyde, and thus a small amount of urate deposits may totally dissolve from tissue samples. The aim of the present study was to identify those steps of the staining procedure that are responsible for the dissolution of urate crystals. We found that the dissolution of urate crystals during the course of staining was caused by the combined effects of haematoxylin staining, treatment with 1% aqueous lithium carbonate solution and dehydration with acetone. As the simplest histological method for the detection of urate crystals, we recommend examining unstained sections (mounted with Canada balsam) of formalin-fixed, paraffin-embedded tissue samples in polarised light. According to our previous study, about two thirds of urate crystals remain detectable on unstaied sections, whereas haematoxylin-eosin stained sections of the same tissue samples (derived from patients with gout) did not contain urate crystals. In the samples where urate crystals could be detected in haematoxylin- eosin stained sections using polarised light, the unstained sections contained much more crystals, which shows that dissolution is greatly decreased on unstained sections.]

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[BACKGROUND - The spontaneous osteoporotic fracture of the sacrum, known as a sacral insufficiency fracture (SIF) was first described as an unrecognized syndrome of the elderly by Laurie, in 1982. Numerous case histories and a few series of cases have been discussed in medical journals; however, none have been reported in Hungary. GOAL - To delineate the leading diagnostic steps in the recognition of SIF and review the therapeutic guidelines. CASE HISTORIES, METHODS - Between January 2009 and the first six months of 2010 11 cases of SIF were diagnosed at the National Center for Spinal Disorders. We examined the clinical aspects of the illness, the radiological modalities, the fracture markings, the pace of recovery and duration. RESULTS - The 11 patients were found to have various SIF predestining etiological factors and the following classic fractures - H-type, unilateral, horizontal, unilateralhorizontal and vertical as well as a bilateral pattern. In cases often not showing obvious clinical symptoms and in cases resulting in conventional radiological examinations of low sensitivity and specificity, we used mapping techniques in setting up the exact diagnosis. CONCLUSION - If we consider SIF from patient history and known risk factors, diagnostic procedure (primer original) may be shortened and a number of unnecessary tests (biopsy) may be avoided.]

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[Recurrent osteoporotic bone fractures are less and less considered “natural”, due to the immense variety of products available for treatment. In order to prevent recurrent fractures, treatment should be started in time, and a careful approach is needed to choose the appropriate treatment, and, if needed, to switch therapy. When choosing the therapeutic approach, we have to decide whether it corresponds to the severity of the osteoporosis and the risk of fracture. In order to do this, we have to consider bone quality, previous fractures, the condition of cortical bones and the mode of action of the selected treatment, in addition to the easily evaluated density value. The aim of this article is to provide practical help for the above mentioned decisionmaking process.]

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[The authors studied the solubility of urate crystals in alcohol, in an 8% aqueous solution of formaldehyde and in acetone, respectively. The urate crystals were least soluble in alcohol. In comparison, the amount of urate crystals decreased in the aqueous solution of formaldehyde, which confirmed the suggestion of our predecessors that tissues suspected to contain urate crystals should be fixed in alcohol. Urate crystals dissolved in greatest amounts in acetone. Acetone is widely used by histological laboratories for dehydration of tissue blocks before embedding them in paraffin, which, in case of fixation in aqueous formaldehyde, contributes to the dissolution of urate crystals. In our earlier studies, we found that dissolution of urate crystals from haematoxylineosin stained sections is caused by the staining of nuclei in haematoxylin, therefore urate crystals are preferably demonstrated in unstained tissue sections.]

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[In our previous study we refuted the thesis that sodium urate crystals are not, or only rarely detectable in formalin-fixed histological samples because they dissolve in the aqueous formalin solution. Our observations indicate that dissolution of urate crystals is primarily caused by haematoxylineosin staining. Undeniably, however, urate crystals are partially dissolved in the aqueous solution of formaldehyde, and thus a small amount of urate deposits may totally dissolve from tissue samples. The aim of the present study was to identify those steps of the staining procedure that are responsible for the dissolution of urate crystals. We found that the dissolution of urate crystals during the course of staining was caused by the combined effects of haematoxylin staining, treatment with 1% aqueous lithium carbonate solution and dehydration with acetone. As the simplest histological method for the detection of urate crystals, we recommend examining unstained sections (mounted with Canada balsam) of formalin-fixed, paraffin-embedded tissue samples in polarised light. According to our previous study, about two thirds of urate crystals remain detectable on unstaied sections, whereas haematoxylin-eosin stained sections of the same tissue samples (derived from patients with gout) did not contain urate crystals. In the samples where urate crystals could be detected in haematoxylin- eosin stained sections using polarised light, the unstained sections contained much more crystals, which shows that dissolution is greatly decreased on unstained sections.]

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[A dogma of histochemistry that seems to be refuted - histological detectability of urate crystals]

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[In medical practice there are a number of “truths etched in stone” that are passed on from textbook to textbook and learned by generations before they become obsolete. This short study aims to eliminate a misbelief from the diagnosis of gout that is related to the histological detectability of urate deposits. According to the generally accepted thesis, urate crystals obtained from patients with gout are dissolved in formalin solution, therefore, tissue samples should be fixated in alcohol. The authors have found that urate crystals can be detected on conventionally mounted, native (unstained) sections, despite formalin fixation, whereas the great majority of urate crystals are dissolved during haematoxylin-eosin staining. Therefore, for the detection of urate crystals the tissue samples should be examined on native, unstained sections.]