Kronvall G, Myhre E
Acta Pathol Microbiol Scand B. 1977 Aug;85(4):249-54. doi: 10.1111/j.1699-0463.1977.tb01970.x.
Optimal conditions for acridine orange staining of air dried and methanol fixed bacteria on glass slides were studied. The pH of the staining buffer did not influence the fluorescence of an S. aureus and an E. coli strain at dye concentrations of 25-50 mg per litre. 81 bacterial strains representing 15 different species were stained with acridine orange under standard conditions, all strains showing orange fluorescence. The pH of the buffer influenced markedly the staining patterns of human cells and tissue materials, as represented by smears of peripheral blood, buccal scrapings, urethral secretions and tracheal exudates. The fluorescence obtained ranged from low intensity green at low pH values to bright orange at neutral and alkaline pH. This variability indicated a possibility of designing conditions for a differential staining method for the detection of bacteria in clinical specimens. The differential staining effect with a low pH in the buffer was confirmed on smears of buccal scrapings, cerebrospinal fluid samples and urethral secretions, showing orange fluorescence of the bacteria present and green-to-yellow fluorescence of background material, cells and tissue debris.
研究了在载玻片上对空气干燥并经甲醇固定的细菌进行吖啶橙染色的最佳条件。在每升25 - 50毫克的染料浓度下，染色缓冲液的pH值对金黄色葡萄球菌和大肠杆菌菌株的荧光没有影响。在标准条件下，用吖啶橙对代表15个不同物种的81株细菌进行染色，所有菌株均呈现橙色荧光。缓冲液的pH值显著影响人体细胞和组织材料的染色模式，以外周血涂片、颊部刮片、尿道分泌物和气管渗出物为例。获得的荧光范围从低pH值时的低强度绿色到中性和碱性pH值时的亮橙色。这种变异性表明有可能设计出用于临床标本中细菌检测的鉴别染色方法的条件。在颊部刮片、脑脊液样本和尿道分泌物涂片上证实了缓冲液低pH值的鉴别染色效果，显示存在的细菌发出橙色荧光，背景材料、细胞和组织碎片发出绿色至黄色荧光。
