Biophysics & Instrumentation

UNIT –I: BASIC LABORATORY INSTRUMENTS

Buffers: Henderson and Hesselbach equation, pka and pkb. Preparation of Buffers.Viscosity: specific, intrinsic and reduced viscosities, viscometers, determination of molecular size and shape through viscosity.

Osmosis: osmosis in relation to molecular size and molecular weight, osmometer, partial, specific volume and diffusion co-efficient, Dialysis, membrane filtration and its application.

Principle and working of: pH meter, Sterilization techniques

UNIT – II: SEPARATION TECHNIQUES

Centrifugation: Types of centrifuge machines, analytical centrifuges, differential centrifuge, and Ultracentrifuge. Basic principles and applications: of gel filtration chromatography, ion exchange and affinity chromatography, gas liquid chromatography (GLC), high pressure/performance liquid chromatography (HPLC). Basic principles of Electrophoresis, Agarose gel and SDS- PAGE, isoelectric focusing, 2D Electrophoresis.

UNIT -III: SPECTROSCOPY, RADIOGRAPHY AND TRACER TECHNIQUES

Spectroscopic techniques: theory and applications of UV, Visible, IR, NMR, Fluorescence, AAS and AES. Principles and applications of tracer techniques in biology: Radioactive isotopes and half life of isotopes, Effect of radiation on biological system; Autoradiography, Cerenkov radiation, Liquid scintillation counter, Dosimetry. Laboratory procedures and safety aspects.

UNIT -IV: MICROSCOPY AND STAINING OF MICROORGANISMS

Microscopy: Principles and microscope application of bright-field and dark-field microscopy, Phase- contrast microscopy, fluorescence microscopy, Electron Microscopy : TEM, SEM and AFM , Confocal microscopy Cytophotometry and flow cytometry. Fixation and staining: Simple staining, negative staining, gram staining, acid fast staining, structural stains (Endospore, capsule and flagella).