《微生物学与免疫学实验》课程教学资源（讲义）细菌的革兰氏染色

微生物学与免疫学实验模块三微生物的形态观察、生理生化反应及鉴定细菌的革兰氏染色模块三

微生物学与免疫学实验 模块三 微生物的形态观察、生理生化反应及鉴定 之 模块三 细菌的革兰氏染色

主要内容常用染色方法的介绍细菌的革兰氏染色方法草兰氏染色方法的原理实验所需要的试剂及器械等操作步骤与注意事项染色结果观案实验目的与要求了解不同的细菌染色方法的原理掌握革兰氏染色方法的操作技术

实验目的与要求  了解不同的细菌染色方法的原理  掌握革兰氏染色方法的操作技术 常用染色方法的介绍 细菌的革兰氏染色方法 革兰氏染色方法的原理 实验所需要的试剂及器械等 操作步骤与注意事项 染色结果观察 主要内容

常用染色方法Negative staining(负染）Negative, indirect, or background staining isachieved by mixing bacteria with an acidicstain such as nigrosin, India ink,or eosin,andirstslidethen spreading out the mixture on a slide toform afilm.The above stains will not penetrateand/stain the bacterial cells due to repulsionbetween the negative charge of the stainsandthe negativelycharged bacterial wall.+++++

Negative staining（负染） Negative, indirect, or background staining is achieved by mixing bacteria with an acidic stain such as nigrosin, India ink, or eosin, and then spreading out the mixture on a slide to form a film. The above stains will not penetrate and stain the bacterial cells due to repulsion between the negative charge of the stains and the negatively charged bacterial wall. 常用染色方法

NegativelyStained Bacillus:Negatively Stained Cocci(A)NegetativeCell(B) Endospore

Negatively Stained Bacillus : (A) Vegetative Cell (B) Endospore Negatively Stained Cocci

常用染色方法Simplestains（简单染色）The use of a single stain or dye tocreatecontrastbetweenthebacteria andthe backgroundisDyerepelledDye aftrected拉referred fo as simple staining.IfsBasic dye (+1 Becterfium (-)Acidic dye(-/Bacterium (--)Call stainedCellclearchiefvalue lies in its simplicity and)Simple stain technique(o) Negative staln techniqueease of use. Simple staining isoffen employed wheninformationaboutcellshape,size,andarrangementisdesired

Simple stains（简单染色） The use of a single stain or dye to create contrast between the bacteria and the background is referred to as simple staining. Its chief value lies in its simplicity and ease of use. Simple staining is often employed when information about cell shape, size, and arrangement is desired. 常用染色方法

常用染色方法Differential stains（差别染色）Differential stainsregctdifferentlywithdifferentkindsofbacteria.e.g.,Gram stain, acid.fast stainClump formAgid-faststain（抗酸染色）：结核分枝杆菌（Mycobacteriumtuberculosis张盆酸酒精脱色16分钟复红加热染5分钟涂玻膜自然干燥镜检美兰复染30流流洗水水冲水CC自然干燥冲中洗洗洗秒

Differential stains（差别染色） Differential stains react differently with different kinds of bacteria. e.g., Gram stain, acid￾fast stain 常用染色方法 涂 痰 膜 自 然 干 燥 复 红 加 热 初 染 5 分 钟 自 然 干 燥 镜 检 流 水 冲 洗 美 兰 复 染 30 秒 5% 盐 酸 酒 精 脱 色 1-3 分 钟 流 水 冲 洗 流 水 冲 洗 Acid 结核分枝杆菌(Mycobacterium tuberculosis) -fast stain (抗酸染色)：

常用染色方法Special stains（特殊染色）Used to color and isolate various structures,such as capsules,endospores,andflagella.FlagellaStainofProteus ShowingPeritrichousArrangementofFlagella

Special stains （特殊染色） Used to color and isolate various structures, such as capsules, endospores, and flagella. 常用染色方法 Flagella Stain of Proteus Showing Peritrichous Arrangement of Flagella

(a)Simple Stains(b)DifferentialStains(c) Special StainsCrystal violetGramstainIndia ink capsule stain ofPurple cells are gram-positive.stain of EschenichiaCryptococcusneoformanscoWRed cells aregram-negative.MethyleneblueAcid-fast stainFlagellar stain of Proteus vuigarisRed cells are acid-fast.AbasicstainwasusedtostainofCorynebactenumBlue cells are non-acid-fast.build upthe flagellaSpore stain, showing spores (red)andvenetative.callsfblue

ShapeArrangementSphericaldiplococcus(pairs)COCCUS(pl.,cocci)streptococcus(chains)staphylococcusCommon Bacterialfrandomorgrapelike clusters)Shapes andmicrococousArrangements(squaregroupsoffourcells)Rod-shapedstreptobacillus(chains)bacillus(pl,bacillnsarcinaSpiral(cubicalpacketsspirillumofeightcells)(pl.,spirilla)Incompletespiralvibrio(pl.,vibrios)Iregularorvariableshapepleomorphic

Common Bacterial Shapes and Arrangements

革兰氏染色VGramStain-ChristianGram.in1884Thegram stainisthemostwidelyusedstainingprocedurein bacteriology.Ifiscalledadifferential stain since it differentiatesbetweengram-positive andgram-negative bacteria.Bacteria which stain purplewiththegramstainingprocedure aretermed gram-positive;thosewhich stain pink are said fo be gram-negative

革兰氏染色  Gram Stain - Christian Gram，in 1884  The gram stain is the most widely used staining procedure in bacteriology. It is called a differential stain since it differentiates between gram-positive and gram-negative bacteria.  Bacteria which stain purple with the gram staining procedure are termed gram-positive; those which stain pink are said to be gram￾negative