2.  현미경의 3 주요 부분  Light source  Lens  Condenser ( 집광기 )  현미경의 3 주요 기능  Magnification ( 배율 )  Resolution ( 해상도, 분해능 )  Contrast ( 대비력 )  Light microscope (LM) Electron microscope (EM) Scanning probe microscope Microscopy

3.  Light (Optical) Microscope  Bright field microscope  Dark field microscope  Phase contrast microscope  Differential interference contrast (DIC) microscope : 3D  Fluorescence microscope  Confocal scanning laser microscope  Electron microscope  Scanning electron microscope (SEM)  Transmission electron microscope (TEM)  Scanning probe microscope ( 주사탐침현미경 )  Scanning tunneling microscope  Atomic force microscope (AFM)

4.  Resolution  Abbé equation d = 0.5λ / n sinθ d : minimum distance between two objects that reveals them as separate entities λ : wavelength n sinθ : numerical aperture (NA, 조리개수 )  λ↓ - light source  NA ↑ - lens, immersion oil  Contrast  Instrumental : phase contrast  Staining

5.  Fixation ( 고정 )  Heat fixation  Chemical fixation chemical fixatives : ethanol, acetic acid, mercuric chloride, formaldehyde, glutaraldehyde  Staining ( 염색 )  Dye : chromophore ( 발색단 ) Basic (cationic) dye : 세포 염색 Acidic (anionic) dye : 바탕 염색 (negative staining) Neutral dye : 지질 등 염색  Mordant ( 매염제 ) iodine, ferrous sulfate, phenol, tannic acid Staining

6.  Basic (cationic) dye methylene blue, crystal violet, safranin, basic fuchsin, gentian violet, methyl violet, malachite green, rosaniline, hematoxylin  Acidic (anionic) dye nigrosin, eosin, rose bengal, erythrosin, picric acid, orantia, acidic fuchsin, orange G  Neutral dye Sudan III (Sudan Black) : lipid Schiff’s reagent (Feulgen procedure) : sugar (DNA) Giemsa stain, Wright tain, Leishman stain Dyes

7.  Staining  Simple staining ( 단순염색 ). size, shape, arrangement. crystal violet, basic fuchsin, methylene blue nigrosin (negative staining)  Differential staining ( 분별염색 ). stain specific cells :. Gram stain : Crystal violet & Safranin Acid-fast stain : Basic fuchsin & Methylene blue Mycobacterium. stain specific structures flagella, endospore, capsule, nuclear stain

8.  Gram stain

9.  Acid-fast stain Mycobacterium tuberculosis mycolic acid in cell wall impenetrable by aqueous stain solutions Koch & Ehrlich simultaneously introduced a stain for M. tuberculosis Ziehl & Neelsen method modifications carbol fuchsin stain, acid alcohol decolorizer, counter stain Muller & Chermock method addition of a surfactant (wetting agent) Kinyoun method Fluorescence microscopy : auramine-rhodamine stain Preliminary diagnosis of active tuberculosis : sputum samples