pewarnaan kuman
DESCRIPTION
pewarnaan bakteriTRANSCRIPT
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Nurul Wiqoyah
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• Living state: Wet Mount Method Hanging drop Preparat
• Fixed state: pewarnaan
• Fiksasi:• Melekatkan spesimen pada obyek glass• Membunuh kuman tanpa merubah morfologinya• Mencegah enzym yang dapat merusak morfologi sel• Sebagai intensifier sehingga kuman mudah di cat
• Macam fiksasi:Secara phisic: pemanasan
Secara chemis: methyl/ethyl alkohol, formalin, asam asetat
•
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• Sifat cat:• Mempunyai : struktur cincin benzena gugus chromofor menentukan warna cat gugus auxochrome menentukan sifat
elektrolit dissosiasi/ kemampuan membentuk salt linkages dengan protein jaringan dll.
Macam cat:
Basic dyes: garam-garam cat yang chromofor (ion-ion cat)nya kation dengan anion: Cl, SO4, asetat, oksalatex. Methylen blue, safranin, crystal violet
Acid dyes: khromofornya anion dengan kation: Na+, K+, Ca+, NH4+
Ex. Fuchsin, acid fuchsin, congo red dll
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• Preparasi smear:
• Smear: lesi, exudate, darah, jaringan atau koloni kuman yang terisolir
Smear fiksasi pewarnaan
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Types of stain
• Simple stains
• Differential stains
• Special stains
• General stains
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Pewarnaan sederhana
• Menggunakan satu macam zat warna• Kuman terwarnai zat warna
• Ex. Methylen blue kuman berwarna biru
Safranin kuman berwarna merah
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Pewarnaan negatif
• Zat warna Iodin Ink
• Kuman tidak terwarnai dengan bakcground gelap
• Kapsul jernih
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Pewarnaan differensial
1. Pewarnaan Gram:
Paling umum digunakan Tehnik pengecatan diagnostik
Cat: crystal violet (primary stain) Lugol (mordan) Alkohol Safranin (counter stain)
Bakteri Gram positif purple
Bakteri Gram negatif merah
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positivenegative
primary stain
mordant
decolorization
counterstain
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Kuman Gram positif Kuman Gram negatif
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2. Pewarnaan tahan asam ZEIHL-NEELSON METHOD
• Pewarnaan differensial untuk bakteri tahan asam kelompok kuman ycobacterium
Prinsip: tergantung pada ketahanan bakteri terhadap dekolorisasi
alkohol asam dinding sel mengandung asam mikolat yang
berikatan dengan polisakarida yang mencegah karbol fuchsin dari sel
Acid fast cells merah
Non Acid fast counter stain
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• Pewarnaan kapsul
Congo Red METHOD
• MUIR’S METHOD
• Carbol Fuchsin• Ethanol• MUIR’S Mordant• Counterstain: Loeffler’s methylen blue
• ANTHONY METHOD:Crystal violet20% CUSO4
Pewarnaan khusus
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Pewaranaan spora• Types:
• Moeller Modified Method Carbol Fuchsin, Ethanol, Loeffer’s Methylen Blue Vegetative cell biru, spora merah
• Pewarnaan Schaeffer fulton− Bahan malachite green, safranin− Spora kuman hijau− Vegetative cell merah− Pewarnaan Bartolomev dan Mittwer
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Pewarnaan NEISSER• Zat warna Neisser AB Neisser C
Melihat granula metakhromatikGranula metakhromatik biru gelapVegetatif sel kuning coklat
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• Pewarnaan flagella
• Pewarnaan Leiffson• Pewarnaan Bailey
mordan Asam tannin