Immunofluorescence staining of paraffin sections: creating DAB staining like virtual digital images using CMYK color conversion

• Дата: 2008
• Автори: Buchynska, L., Kashuba, E., Szekely, L.
• Формат: Стаття
• Мова: English
• Опубліковано: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2008
• Назва видання: Experimental Oncology
• Теми: Technical advance
• Онлайн доступ: http://dspace.nbuv.gov.ua/handle/123456789/139927
• Назва журналу: Digital Library of Periodicals of National Academy of Sciences of Ukraine
• Цитувати: Immunofluorescence staining of paraffin sections: creating DAB staining like virtual digital images using CMYK color conversion / L. Buchynska, E. Kashuba, L. Szekely // Experimental Oncology. — 2008. — Т. 30, № 4. — С. 327–329. — Бібліогр.: 14 назв. — англ.

Crystal violet treatment of formalin fixed paraffin embedded tissue slides greatly reduces the endogenous autofluorescence, and allows immunofluorescence (IF) staining with FITC or Alexa488 conjugated antibodies. Using cold CCD camera to capture the fluorescence images makes this staining method very sensitive. Here we show that combination of IF with the simultaneous recording of crystal violet induced red and Hoechst 33258 induced blue fluorescence permits the localization of the IF signal over a cytoplasmic: nuclear red:blue stain that visualizes the microscopic anatomy of the underlying tissue. To make the visual interpretation of the IF staining easier for microscopists, who are used to DAB staining over weak hematoxilin-eosin background, we created a simple color conversion procedure that turns the captured three-color fluorescence RGB (red, green, blue) images over a black background into four color CMYK (cyan, magenta, yellow, key color (black)) images.