Name: ANN NIGKE 6 BARRE Section: _ BAN) A-N Date: _hPe ob, 2023 er | At the end of the laboratory period, the student should be able to: 1. perform the Gram-staining procedure; 2. identify the Gram-staining reaction; and 3. distinguish the fundamental shapes of bacteria. nero | Gram-staining is one of the most basic staining procedures done in the study of Microorganisms. It is a differential stain that groups organisms into gram-positive and gram- Pegative organisms and can aid in the basic identification of organs It aise demonstrates the ic shape of the organism being studied. The following rules will help the student remember Gram-staining reaction of the more important bacteria: ; Corynebacterium, Mycobacterium, aerobic spore- formers (Clostridium). Veillonella, and Branbamella. 1. All bacilli are gram-negative except formers (Bacillus), and anaerobic spore“ 2. All cocci are gram-positive except Neisseria,> 6, Flood the smear with 95% alcohol and let it . ere a ean for 15 to 30 seconds. Repeat this step 7. Wash with water. g. Counter-stain with safranin for 30 seconds. 9, Air dry and examine in the microscope using oil immersion objective. Illustrated Gram-staining procedure @ Iodine 4 KEY ] © Crystal violet / J © Alcohol —— ~ e © Safranin Gram-positive Gram-negative Qroviication of Application @ Alcohol wash @ Application crystal violet of iodine (decolorization) of safranin (purple dye) (mordant) (counterstain) 1. Give the Gram-staining reaction and morphology (shape) of the following bacteria. oe WER Gram stain reaction: Gram gosttwe aS ANG baderia ays Aa ws ia Says gue Set Morphology/Shape: Spores, of Coca ~~ Aut Gram stain reaction: Gram wegaine as Ake backria tums pink ea SA ot hology/Shape: Raa or padi Xe Sy jorphology/Shape: Rod or bau NS. Gram stain reaction: Gram Negative as he Imcuria turns A Morphology/Shape: ted wr bitiMicrobiology and Parasitology: A Textbook and Laboratory Manual for the Health Sciences IL. Indicate the function of the different reagents used in Gram-staining. Til. Expected Result Reagent Punction Gram-positive Gram-negative roan oS ae els appear purple agetat gin Crystal violet bine ae in i mas and "Fives ane ol an iz A eh ANG A al HAE (1 Gram's iodine | obgeax purge yk b etd {mci ty wiritg 184 lh ve 00) [trea ee 95% Alcohol | ip SMliaRNt Paccdune | nae - Lan ™ parte colaeless Capea as fiom | | sekaineA Ye ge : satranin | ant Temes dtig cH agRot | coward and | a dar pete wor shain weds Differentiate gram-positive cell wall from gram-negative cell wall. Features Gram-positive Cell Wall | Gram-negative Cell Wall Yack quter membrane at Shiroundd Nie times ANN peat Peptidoglycan wt We cigs Cong sung of gps) LOE Ria of oR ter Complexity me and han! seagate and phespongi » {Mao ayer * tones a Wwe pertmiage — | 1 Yd, ore) ay ack Teichoic acid o aa paitive pa other er wind ik howd 8 Hide geet dag NYcan yer | Widwy Ala vaccha roi Lipopalysaccharde ed He Plasma, ded on te cute tated df ‘prant um ran iar! Visti lwnony) wo : Endotoxin mp omental: | Heep cauhaice nine Byam ergohive Watson