Charles J. Churukian

Improved Puchtler's Congo Red Method for Demonstrating Amyloid

Abstract This paper describes a modification of Puchtlersa lkaline Congo red method for demonstrating amyloid in human tissues. Special emphasis is given to the stability of the new staining solution, which has much greater stability than the one reported by Puchtler. This was accomplished by reducing the amount of sodium chloride and alcohol in the solution. The method is less time consuming than Puchtlers method because we found it unnecessary to pretreat tissue with alkaline sodium chloride in alcohol. Tissue sections are stained with a modified Weigert iron hematoxylin and placed into the alkaline Congo red solution for 20 min. They are then dehydrated rapidly in alcohol, cleared in xylene, and mounted with synthetic resin. Staining results obtained with this method are consistent and reliable. (The J Histotechnol 23:139, 2000)

Alcian blue pyridine variant–a superior alternative to alcian blue 8GX: staining performance and stability

We compared the staining performance, dye content, solubility, and visual absorption maximum of two batches of alcian blue pyridine variant and of five batches of alcian blue 8GX (C.I. 74240). Whenever possible, we also compared results to those obtained with the same dye batches produced at an earlier date to provide information concerning dye stability. Both alcian blue pyridine variant batches were of high dye content, stable, of satisfactory solubility, and performed well in both the routine Mowry mucin stain and in the critical electrolyte concentration (CEC) stain. Of the five alcian blue 8GX samples, some were also of appropriate dye content, were sufficiently stable, and gave good staining in the two procedures. Two batches, however, were unstable, and three batches were unsatisfactory in staining performance and solubility in the CEC stain. Consequently alcian blue pyridine variant is a superior substitute for alcian blue 8GX.

Staining Problems with Eosin Y: A Note from the Biological Stain Commission

In 1980, eosin Y was the certified dye with which technologists encountered most problems. The specific problem most frequently brought to the attention of the Biological Stain Commission was that solutions of eosin Y formed a precipitate and failed to stain cytoplasm red when used as a counterstain to hematoxylin.

A Simple Colloidal Silver Method (Autometallographic Technique) for Demonstrating Inorganic Mercury in Brain Sections

Abstract Standard methods for mercury measurement estimate total mercury burden in tissues but are not informative about cellular localization, for which histochemical techniques are needed. Because the main target of the toxic effects of mercury is the nervous system, it is of particular interest to localize its reservoir within the brain. This paper presents a simple, autometallographic method for demonstrating inorganic mercury in paraffin sections of brain by utilizing a buffered colloidal silver solution that contains hydroquinone. Formalin fixed, paraffin processed brain sections are cut at 7 μm and hydrated with deionized water. The sections are placed in a buffered colloidal silver nitrate solution containing hydroquinone and kept in a 37°C oven for 18 to 20 min. They are then washed immediately in hot running water and rinsed in deionized water. Following treatment with 2% sodium thiosulfate for 1 min, they are rinsed in deionized water and counterstained with nuclear fast red for 3 min or with cresyl violet acetate for 5 min. After the slides are rinsed in deionized water, they are dehydrated in alcohol, cleared in xylene, and mounted with synthetic resin. Mercury stains black and the results are consistent and reliable. (The J Histotechnol 23:337, 2000)

The Importance of Fluorochrome Staining for the Identification of Mycobacterium Tuberculosis

Abstract A patient with a history of chronic obstructive pulmonary disease was hospitalized after suffering second-degree burns to her mouth, nostrils, and the right side of her face. She was rushed to the hospital, immediately intubated, and sedated with Ativan. Her chest x-ray confirmed changes consistent with chronic obstructive pulmonary disease. Later chest x-ray confirmed showed her to have a calcified granuloma of the right base posteriorly as well as increased changes bilaterally. She died a few days later. At autopsy, granulomas were found in the right lower lobe of her lung. Special stains for acid-fast bacteria were performed. The Ziehl-Neelsen carbol-fuchsin method revealed only a few mycobacteria whereas Churukians modification of Truants fluorochrome stain showed large numbers of the microorganism. (The J Histotechnol 29:41, 2006) Submitted May 20, 2005; accepted with revisions July 26, 2005.

Microwave Methods for Demonstrating Melanin

Abstract Microwave modifications of Churukians ammoniacal silver, Lillies melanin bleach, Lillies ferrous ion uptake, Lillies Nile blue A, Schmorls for reducing substances, and the Giemsa stain are described and recommended for use in demonstrating melanin. The use of the microwave oven reduces the time required to perform these methods without compromising the quality of the staining results. In fact, the results obtained with these methods are usually better than those obtained with the conventional nonmicrowave techniques. Even though these methods are not as specific or sensitive as the monoclonal antibodies HMB-45 and melanin A and the polyclonal antibody S-100, they are still useful for demonstrating melanin in malignant melanomas. (The J Histotechnol 26:239, 2003)