Report from the President

Although the original Commission on Standardization of Biological Stains was first organized in 1921, it was not until 1944 that this was incorporated as an independent, nonprofit organization known as the Biological Stain Commission (see Clark 1974). The certification of dyes, as indicated by special labels purchased by manufacturers or vendors for attachment to the dye containers, originated with the parent organization and has continued to this day. The objectives of the Biological Stain Commission (BSC) are 1) to identify and standardize the content and performance of dyes and dye preparations used in staining biological tissues and products, 2) to issue labels of certification to companies that buy these to inform consumers that their certified dyes meet the specifications of the BSC, 3) to carry out and to support investigations on dyes and their performance, 4) to publish scientific data concerning biological stains and their use, and 5) to maintain, through scientific meetings and correspondence, an active “dialogue” among scientific and industrial personnel concerned with biological stains. The present report summarizes Commission activity and some of the changes that have occurred during the past five years.     

Report from the President the Biological Stain Commission: Its Goals, Its Past and Its Present Status

This is a brief overview of the goals, evolution, and present status of the Biological Stain Commission. The main function of the Commission is the testing and certification of dye batches intended for biological applications. The testing is supported by charges made for batch testing and by the sale of certification labels affixed to individual dye containers. Submission of dyes for testing is voluntary, depending on the cooperation of the companies who sell them and the consumers who buy them. The supportive role of the University of Rochester School of Medicine and Dentistry—both past and present—is not well known and should be. Increasingly federal regulations affect the production, availability, and cost of dyes. Commission income from the sale of labels has decreased in recent years. Continuation of its work requires changes that will produce more income. Much dye is now sold in solutions instead of dry powders. The value of using Stain Commission certified dyes whenever possible is illustrated by the case of basic fuchsin. Years ago this dye was a mixture. Most basic fuchsin now marketed consists mainly of either pararosanilin (Colour Index No. 42500) or rosanilin (C.I. No. 42510). The Biological Stain Commission discovered that some certified batches of both pararosanilin and rosanilin sold as “basic fuchsin” had incorrect C.I. numbers on the labels. Sometimes that caused failure of the aldehyde fuchsin stain. Unless made with pararosanilin, aldehyde fuchsin does not stain pancreatic islet B-cells, elastic fibers, and hepatitis B surface antigen in unoxidized sections. Mislabelling by packagers may interfere with other applications of pararosanilin and rosanilin. The Commission acted to publicize and correct this problem. Biological Stain Commission publications help educate microscopists and histotechnologists about dyes and their best use. Stain Commission representatives from member scientific societies provide valuable input about changes in the availability and quality of such dyes as hematoxylin and others; they also provide useful feedback to their societies about dye problems. Each new generation of biologists and histotechnologists should be taught the importance of using only Stain Commission certified stains when available. They should be taught also to notify the Stain Commission whenever they experience problems with any certified dye.     

News from the Biological Stain Commission No. 11

The 11th issue of News from the Biological Stain Commission (BSC) provides our first impressions of the REACH and ECHA programs. We intend to give a more thorough account of what these important programs actually mean in later editions of News from the Biological Stain Commission. Under the heading of Regulatory Affairs, the Biological Stain Commission’s International Affairs Committee presents information from the opening session of the meeting of the International Standards Organization ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 2-4 June 2010 in Seoul, Republic of Korea.     

News from the Biological Stain Commission No. 11

The 11th issue of News from the Biological Stain Commission (BSC) provides our first impressions of the REACH and ECHA programs. We intend to give a more thorough account of what these important programs actually mean in later editions of News from the Biological Stain Commission. Under the heading of Regulatory Affairs, the Biological Stain Commission's International Affairs Committee presents information from the opening session of the meeting of the International Standards Organization ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 2-4 June 2010 in Seoul, Republic of Korea.     

Purity of commercial non-certified European samples of Pyronin Y

Summary The purity of six European non-certified samples of Pyronin Y was compared with that of two American samples certified by the Biological Stain Commission. The methods used were spectrophotometry and a Methyl Green-Pyronin staining test (both as applied by the Biological Stain Commission), thin layer chromatography, mass spectrometry, determination of pH, and content of some electrolytes. It was found that none of the European batches of Pyronin Y passed the complete test as prescribed by the Biological Stain Commission. Their dye content was uniformly low (between 5 and 19%). Furthermore, thin layer chromatography and mass spectrometry revealed that two of the dye samples contained no Pyronin Y or only traces.It is concluded that assessment of an unknown sample of a dye labelled Pyronin Y should be initiated with thin layer chromatography. The pH and content of electrolytes in an aqueous solution of the dye should also be determined in order to obtain reproducible staining results. Finally, the value of the work performed by the Biological Stain Commission is underlined, although more sophisticated methods are necessary for testing the purity of dyestuffs.     

News from the Biological Stain Commission No. 13

Abstract

In the 13^th issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission’s International Affairs Committee presents information from the first plenary meeting of the International Standards Organization ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 17–19 October 2011 in Las Vegas, Nevada.     

News from the Biological Stain Commission No. 15

In the 15^th issue of News from the Biological Stain Commission (BSC), under the heading of Regulatory affairs, the Biological Stain Commission’s International Affairs Committee presents information from the plenary meetings of the International Standards Organization ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on August 22–24, 2012 in Berlin, Germany. An additional discussion of the use of food dyes in India also is included.     

News from the Biological Stain Commission,No. 17

In the 17^th issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission’s International Affairs Committee presents information from the 20^th meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on October 15 ? 17, 2014 in Toronto, Canada, and from the 29^th meeting of CEN/TC 140 In vitro diagnostic medical devices held on February 3, 2015 in Berlin, Germany.     

News from the BSC No. 8

Abstract

In the 8th and following issues of News from the Biological Stain Commission (BSC), under the heading of Regulatory affairs, the BSC's International Affairs Committee will present information from a meeting held in Ghent, Belgium on 15–18 June 2009 concerning the progress achieved by the International Standards Organization Committee ISO/TC 212 Clinical Laboratory Testing and in Vitro Diagnostic Test Systems since the last meeting held in Vancouver, Canada in 2008. A note on the meaning and significance of E numbers found on the labels of foodstuffs and beverages sold for human consumption concludes this edition of News from the Biological Stain Commission.     

News from the Biological Stain Commission,No. 16

Abstract

In the 16^th issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission’s International Affairs Committee presents information from the 28^th meeting of CEN/TC 140 In vitro diagnostic medical devices held on October 23, 2013 in Berlin, Germany. Information is also presented from the 19^th meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on October 19 ? 21, 2013 in Singapore.     

Progress in the Standardization of Stains Research on the Chemistry of Dyes

One of the services that the Stain Commission has been able to perform for biologists is in assisting them to identify dyes of unknown composition with which desired staining effects have been secured. This enables them both to obtain new supplies of the same dyes and to publish their results in such a form that other biologists may duplicate them.     

News from the Biological Stain Commission No. 7

Abstract

In this issue of News from the Biological Stain Commission (BSC), under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from a meeting held in Berlin by the International Standards Organization ISO/TC 212/WG 1, “Quality and Competence in the Medical Laboratory,” on 11–12 December 2008. After this, we turn again to problems with impure dyes and find that solvent dyes are impure even for non-biological use.     

Special Symposium Biomarkers—The New Frontier in the Pathology of Invasive and Preinvasive Neoplasias

The papers in this symposium were presented at the 1996 Annual Meeting of the Biological Stain Commission. The authors hope that the discussion of biomarkers in the papers included in this issue and the literature reviewed in them will be useful to diagnostic pathologists as well as to investigators studying neoplastic processes in general.     

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.     

Only aldehyde fuchsin made from pararosanilin stains pancreatic B cell granules and elastic fibers in unoxidized microsections: problems caused by mislabelling of certain basic fuchsins

The most distinctive property of aldehyde fuchsin is its staining of certain nonionic proteins and peptides in unoxidized cells and tissues. These substances include granules of pancreatic islet B cells, elastic fibers and hepatitis B surface antigen. Aldehyde fuchsin made from two different basic fuchsins, each certified by the Biological Stain Commission and labelled C.I. (Colour Index) No. 42500 (pararosanilin), did not stain pancreatic B cells at all. Stain Commission's records and retesting showed that each of the "faulty" basic fuchsins was not pararosanilin, but rosanilin, whose Colour Index number is 42510. These basic fuchsins were labelled with the wrong Colour Index number when packaged. Additional basic fuchsins were coded by V.M.E. and tested by R.W.M. for their capacity to make satisfactory aldehyde fuchsins. Only certain of these aldehyde fuchsins stained unoxidized pancreatic islet B cells. The same aldehyde fuchsins stained elastic fibers strongly. Each basic fuchsin whose aldehyde fuchsin was judged satisfactory proved to be pararosanilin. Aldehyde fuchsin solutions made from other basic fuchsins stained elastic fibers only weakly and did not stain pancreatic B cells at all in unoxidized sections. Each basic fuchsin whose aldehyde fuchsin was unsatisfactory proved to be rosanilin. It appears that only aldehyde fuchsin made from pararosanilin stains unoxidized pancreatic B cell granules dependably. We found that basic fuchsins from additional lots of Commission-certified pararosanilin and rosanilin were also labelled with incorrect Colour Index numbers when packaged. Steps were taken to prevent recurrences of such mislabelling which has made it difficult until now to correlate differences in the properties of pararosanilin and rosanilin. A table is provided of all basic fuchsins that have been certified by the Biological Stain Commission since 1963 when they began the practice of subdesignating basic fuchsins according to whether they are pararosanilins or nonpararosanilins. The consumer can readily determine from the certification number on the label the correct subdesignation of any Commission-certified basic fuchsin listed here. Until now, mislabelling of some lots of pararosanilin as rosanilin and vice-versa has confused and frustrated the users of basic fuchsins in other applications such as the carbol fuchsin staining of tubercle bacilli and certain cytochemical tests, e.g. esterase and acid phosphatase, that utilize hexazotized pararosanilin as a coupling reagent. Consumers experiencing trouble with any Commission-certified dye should look to the Biological Stain Commission for help. This is an important reason for purchasing, whenever possible, only Biological Stain Commission certified dyes.     

News from the Biological Stain Commission No. 12

Abstract

In this 12^th issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission’s International Affairs Committee presents information from the meetings of ISO/TC 212/WG 1 Quality and competence in the medical laboratory and ISO/TC 212/WG 3 In vitro diagnostic products both held on 2 – 3 June 2010, plus information on the second plenary meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 4 June 2010. All meetings took place in Seoul, Republic of Korea. Finally, information is provided concerning the 25^th meeting of CEN/TC 140 In vitro diagnostic medical devices held on 23 June 2010 in Berlin, Germany.     

Edmund Keffer Kline 1894-1976

It is with deep regret that the friends of Dr. E. K. Kline learned of his death on January 26th, 1976. Dr. Kline served with the Biological Stain Commission for many years, joining the Board of Trustees as representative of the American Public Health Association shortly after the reorganization of the Commission during the latter stages of World War II. Dr. Kline served as Vice-president of the Commission from 1963 to 1966, and as President from 1966 to 1969. His warm good nature and sound judgement combined effectively to help advance the goals of the Commission throughout his period of service, and his absence will be keenly felt.     

News from the Biological Stain Commission no. 12

In this 12(th) issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from the meetings of ISO/TC 212/WG 1 Quality and competence in the medical laboratory and ISO/TC 212/WG 3 In vitro diagnostic products both held on 2 - 3 June 2010, plus information on the second plenary meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 4 June 2010. All meetings took place in Seoul, Republic of Korea. Finally, information is provided concerning the 25(th) meeting of CEN/TC 140 In vitro diagnostic medical devices held on 23 June 2010 in Berlin, Germany.     

A method for determining identity and relative purity of carmine, carminic acid and aminocarminic acid

Carmine is one of the few dyes currently certified by the Biological Stain Commission that is not assayed for dye content. Existing assay methods are complex and do not differentiate the three cochineal derivatives carmine, carminic acid and aminocarminic acid. The latter dye is relatively new to the food trade as an acid-stable red colorant and may eventually enter the biological stains market. The assay proposed here is a two-step procedure using quantitative spectrophotometric analysis at high pH (12.5-12.6) followed by a qualitative scan of a low pH (1.90-2.10) solution. Carmine is distinct at high pH, and the remaining dyes are easily distinguished at low pH. Four instances of mislabeling are documented from 18 commercial products, but the mislabeled dyes were not certified dyes. Samples from nearly all lots of carmine certified by the Biological Stain Commission from 1920 to 2004 proved to be carmine, but they varied widely in dye content. Batches from 1920 through the 1940s were significantly richer in dye content. Variability has been extreme since 2000, and most of the poorest lots have been submitted since 1990.     

A method for determining identity and relative purity of carmine,carminic acid and aminocarminic acid

Carmine is one of the few dyes currently certified by the Biological Stain Commission that is not assayed for dye content. Existing assay methods are complex and do not differentiate the three cochineal derivatives carmine, carminic acid and aminocarminic acid. The latter dye is relatively new to the food trade as an acid-stable red colorant and may eventually enter the biological stains market. The assay proposed here is a two-step procedure using quantitative spectrophotometric analysis at high pH (12.5–12.6) followed by a qualitative scan of a low pH (1.90–2.10) solution. Carmine is distinct at high pH, and the remaining dyes are easily distinguished at low pH. Four instances of mislabeling are documented from 18 commercial products, but the mislabeled dyes were not certified dyes. Samples from nearly all lots of carmine certified by the Biological Stain Commission from 1920 to 2004 proved to be carmine, but they varied widely in dye content. Batches from 1920 through the 1940s were significantly richer in dye content. Variability has been extreme since 2000, and most of the poorest lots have been submitted since 1990.