© Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALEMicroscopy OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION and © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTIONBacterialNOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC Staining© Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION TechniquesNOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION icroscopy had its beginnings with the ability of Robert Hooke and Anton van Leeuwenhoek to see small objects that often M© Jones & Bartlettwere invisible Learning, to the LLC naked eye. In fact, Leeuwenhoek© Jones & wasBartlett Learning, LLC theNOT first FOR to see SALE bacteria OR DISTRIBUTIONwith his “homemade” microscopesNOT (hand FOR lenses). SALE OR DISTRIBUTION Since then, great strides have been made in microscope design, magnifi- cation, and resolution. Today’s light (compound) microscope, which uses visible light to © Jones & Bartlettmagnify Learning, and resolve LLC objects, is a core instrument© Jones of &many Bartlett microbiology Learning, LLC NOT FOR SALEresearch OR DISTRIBUTIONlabs. Brightfield microscopy, introducedNOT FOR in SALEExercise OR ,DISTRIBUTION is what you will use with your light microscope in many of the laboratory exercises in this lab manual. Bacterial cells are not easy to see with brightfield microscopy because (1) the cells are© extremely Jones & Bartlettsmall and Learning, (2) most appearLLC colorless against ©a Jones & Bartlett Learning, LLC bright background. Although your light microscope will provide the res- NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION olution and magnification to see them, there still is a contrast problem to solve. This is accomplished by using a colored chemical stain that imparts color to bacterial cells or structures (Exercise ) or to the background (Exercise ). Such simple staining procedures provide the contrast needed to© carryJones out & Bartlettcell measurements Learning, LLCand observations (Table© A ).Jones In addition, & Bartlett Learning, LLC differentialNOT FOR SALE staining OR DISTRIBUTIONprocedures, which use two contrastingNOT FOR colored SALE OR DISTRIBUTION stains, will allow you to separate bacteria into one of two basic groups (Exercise ) and to visualize bacterial structures such as endospores and capsules (Exercise ). © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC. NOT FOR SALE OR DISTRIBUTION.5054 © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC TABLE NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION A Types and Uses for Staining © JonesTYPE & OF Bartlett STAINING Learning,PURPOSE LLC © Jones & Bartlett Learning, LLC To measure cell size NOT FORSimple SALE OR DISTRIBUTIONTo determine cell shape (bacillus, coccus,NOT spiral) FOR SALE OR DISTRIBUTION (uses one stain) To determine cell arrangements To measure cell size and determine cell shape and arrangements Differential To separate bacteria into groups (Gram positive and negative) (uses two contrasting stains) To visualize cell structures (endospores, capsules) © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION L earning Objectives When you have completed the exercises in Part , you should be capable of: © Jones & Bartlett Learning, LLC• Operating the light microscope© Jones correctly & Bartlett in order Learning, to measure andLLC observe living NOT FOR SALE OR DISTRIBUTIONand dead microorganisms.NOT FOR SALE OR DISTRIBUTION • Preparing a bacterial smear. • Calculating bacterial cell size and determining bacterial cell shape and arrange- ment using simple stains. • Completing the Gram stain to correctly identify gram-positive and gram-negative © Jonesbacteria. & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR• Identifying SALE OR(1) bacterial DISTRIBUTION spores and capsules using special NOTdifferential FOR stains SALE and OR DISTRIBUTION (2) bacterial motility. © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC. NOT FOR SALE OR DISTRIBUTION.5054.