cover page as mentioned below: below: mentioned Youas arepage instructedcover the to updateupdate to the coverinstructed pageare asYou mentioned below: Increase the font size of the Course Name. Name. 1. IncreaseCourse the theof fontsize sizefont ofthe the CourseIncrease 1. Name. use the following as a header in the Cover Page. Page. Cover 2. the usein the followingheader a as as a headerfollowing the inuse the 2. Cover Page. ALAGAPPAUNIVERSITY UNIVERSITYALAGAPPA [Accredited with ’A+’ Grade by NAAC (CGPA:3.64) in the Third Cycle Cycle Third the in (CGPA:3.64) [AccreditedNAAC by withGrade ’A+’’A+’ Gradewith by NAAC[Accredited (CGPA:3.64) in the Third Cycle and Graded as Category–I University by MHRD-UGC] MHRD-UGC] by University and Category–I Graded as as Graded Category–I and University by MHRD-UGC] M.Sc. [Botany] 003 630 – KARAIKUDIKARAIKUDI – 630 003 346 13 EDUCATION DIRECTORATEDISTANCE OF OF DISTANCEDIRECTORATE EDUCATION BIOLOGICAL TECHNIQUES IN BOTANY I - Semester BOTANY IN TECHNIQUES BIOLOGICAL M.Sc. [Botany] 346 13 cover page as mentioned below: below: mentioned Youas arepage instructedcover the to updateupdate to the coverinstructed pageare asYou mentioned below: Increase the font size of the Course Name. Name. 1. IncreaseCourse the theof fontsize sizefont ofthe the CourseIncrease 1. Name. use the following as a header in the Cover Page. Page. Cover 2. the usein the followingheader a as as a headerfollowing the inuse the 2. Cover Page. ALAGAPPAUNIVERSITY UNIVERSITYALAGAPPA [Accredited with ’A+’ Grade by NAAC (CGPA:3.64) in the Third Cycle Cycle Third the in (CGPA:3.64) [AccreditedNAAC by withGrade ’A+’’A+’ Gradewith by NAAC[Accredited (CGPA:3.64) in the Third Cycle and Graded as Category–I University by MHRD-UGC] MHRD-UGC] by University and Category–I Graded as as Graded Category–I and University by MHRD-UGC] M.Sc. [Botany] KARAIKUDI – 630 003 003 630 – KARAIKUDIKARAIKUDI – 630 003 BIOLOGICAL TECHNIQUES IN BOTANY EDUCATION DIRECTORATEDISTANCE OF OF DISTANCEDIRECTORATE EDUCATION I - Semester ALAGAPPA UNIVERSITY [Accredited with ‘A+’ Grade by NAAC (CGPA:3.64) in the Third Cycle and Graded as Category–I University by MHRD-UGC] (A State University Established by the Government of Tamil Nadu) KARAIKUDI – 630 003 Directorate of Distance Education M.Sc. [Botany] I - Semester 346 13 BIOLOGICAL TECHNIQUES IN BOTANY Author: Rahul Dev Ambedkar, Assistant Professor, Sri Aurobindo College, Delhi Units (1-14) "The copyright shall be vested with Alagappa University" All rights reserved. No part of this publication which is material protected by this copyright notice may be reproduced or transmitted or utilized or stored in any form or by any means now known or hereinafter invented, electronic, digital or mechanical, including photocopying, scanning, recording or by any information storage or retrieval system, without prior written permission from the Alagappa University, Karaikudi, Tamil Nadu. Information contained in this book has been published by VIKAS® Publishing House Pvt. Ltd. and has been obtained by its Authors from sources believed to be reliable and are correct to the best of their knowledge. However, the Alagappa University, Publisher and its Authors shall in no event be liable for any errors, omissions or damages arising out of use of this information and specifically disclaim any implied warranties or merchantability or fitness for any particular use. Vikas® is the registered trademark of Vikas® Publishing House Pvt. Ltd. VIKAS® PUBLISHING HOUSE PVT. LTD. E-28, Sector-8, Noida - 201301 (UP) Phone: 0120-4078900 • Fax: 0120-4078999 Regd. Office: 7361, Ravindra Mansion, Ram Nagar, New Delhi 110 055 • Website: www.vikaspublishing.com • Email: [email protected] Work Order No. AU/DDE/DE1-238/Preparation and Printing of Course Materials/2018 Dated 30.08.2018 Copies - 500 SYLLABI-BOOK MAPPING TABLE Biological Techniques in Botany Syllabi Mapping in Book BLOCK-1: MICROSCOPY Unit-1: Principles, image formation and applications of Light, Unit 1: Microscopy: Polarizing Microscpy. An Introduction Unit-2: Transmission and Scanning electron microscopes - (Pages 1-10); Confocal Microscopy, Phase Contrast Microscopy. Unit 2: Electron Microscope, Confocal Photomicrography- Digital imaging. Microscopy and Photomicrography Unit-3: Microscopic measurements- Micrometers – Ocular, Stage, (Pages 11-36); Haemocytometer and Camera lucida. Unit 3: Microscopic Measurements (Pages 37-47) BLOCK-2: MICROTOMY Unit-4: Microtomes- Rocking, Rotary, Sledge and Ultra Unit 4: Microtomes microtomes and their uses. (Pages 48-61); Unit-5: Material preparation techniques for microtome sectioning Unit 5: Microtome Sectioning - fixatives, dehydrating agents, killing, fixing methods. (Pages 62-74); Unit-6: Stains and their uses and staining of plant tissues- Clearing, Unit 6: Stains and their Uses Mounting and mountants. (Pages 75-88) BLOCK-3: HISTOCHEMICAL STUDIES Unit-7: Histochemical techniques – staining of Proteins, Unit 7: Histochemical Techniques Carbohydrates, Lipids and enzymes. (Pages 89-99); Unit-8: Microslide preparation—Whole mounts, Smears, Squashes. Unit 8: Micro Slide Preparation Unit-9: Sectioning of Biological specimens- Free hand, Hand (Pages 100-110); microtome, sludge and rotary microtome sectioning, Embedding Unit 9: Sectioning of Biological methods. Specimens Unit-10: Dewaxing and staining of the sections, fixing coverslips (Pages 111-123); and ringing. Unit 10: Dewaxing and Staining (Pages 124-142) BLOCK-4: MACERATION, SEPARATION TRACER AND BLOTTING TECHNIQUES Unit-11: Maceration technique- PAGE, SDS – PAGE and Agarose Unit 11: Maceration Technique gel electrophoresis. Isoelectric focusing. 2D Electrophoresis. (Pages 143-171); Unit-12: Separation technique -Ultracentrifugation- TLC, HPLC, Unit 12: Separation Technique HPTLC, FPLC, GC, MS, MALDI Tof. (Pages 172-220); Unit-13: Tracer techniques- Principles and applications of Unit 13: Tracer Techniques radioactive isotopes, Autoradiography and Liquid scintillation (Pages 221-235); spectrometry. Unit 14: Blotting Techniques Unit-14: Blotting techniques -Principles and techniques of (Pages 236-263) Southern, Northern and Western blotting techniques and hybridization. Principles and applications of PCR, RFLP, RAPD, AFLP and DNA fingerprinting. CONTENTS INTRODUCTION BLOCK I: MICROSCOPY UNIT 1 MICROSCOPY: AN INTRODUCTION 1-10 1.0 Introduction 1.1 Objectives 1.2 Concept of Economic Growth and Development 1.3 Obstacles to Development and Characteristics of Less Developed Countries 1.3.1 Characteristics of Less Developed Countries 1.4 Answers to Check Your Progress Questions 1.5 Summary 1.6 Key Words 1.7 Self Assessment Questions and Exercises 1.8 Further Readings UNIT 2 ELECTRON MICROSCOPE, CONFOCAL MICROSCOPY AND PHOTOMICROGRAPHY 11-36 2.0 Introduction 2.1 Objectives 2.2 Transmission and Scanning Electron Microscope 2.2.1 Scanning Electron Microscope 2.3 Introduction to Confocal Microscopy 2.4 Phase Contrast Microscopy 2.5 Photomirography- Digital Imaging 2.6 Answers to Check Your Progress Questions 2.6 Summary 2.7 Key Words 2.8 Self Assessment Questions and Exercises 2.9 Further Readings UNIT 3 MICROSCOPIC MEASUREMENTS 37-47 3.0 Introduction 3.1 Objectives 3.2 Micrometers 3.2.1 Stage Micrometer 3.2.2 Ocular Meter 3.3 Haemocytometer and Camera Lucida 3.4 Answers to Check Your Progress Questions 3.5 Summary 3.6 Key Words 3.6 Self Assessment Question and Exercises 3.7 Further Readings BLOCK II: MICROTOMY UNIT 4 MICROTOMES 48-61 4.0 Introduction 4.1 Objectives 4.2 Microtomes 4.2.1 Cambridge Rocking Microtome 4.2.2 Rotary Microtome 4.2.3 Sledge Microtome 4.2.4 Ultramicrotome 4.3 Answers to Check Your Progress Questions 4.4 Summary 4.5 Key Words 4.6 Self Assessment Question and Exercises 4.7 Further Readings UNIT 5 MICROTOME SECTIONING 62-74 5.0 Introduction 5.1 Objectives 5.2 Microtome Section and Section Cutting Process 5.3 Material Preparation Techniques for Microtome Sectioning 5.3.1 Fixation 5.3.2 Dehydration 5.3.3 Fixing Agent 5.4 Answers to Check Your Progress Questions 5.5 Summary 5.6 Key Words 5.7 Self Assessment Questions and Answers 5.8 Further Readings UNIT 6 STAINS AND THEIR USES 75-88 6.0 Introduction 6.1 Objectives 6.2 Stain and Staining - Staining of a Plant Tissues 6.2.1 Clearing (Dealcoholization) 6.2.2 Mounting 6.2.3 Mountants 6.3 Answers to Check Your Progress Questions 6.4 Summary 6.5 Key Words 6.6. Self Assessment Questions and Exercises 6.7 Further Readings BLOCK III: HISTOCHEMICAL STUDIES UNIT 7 HISTOCHEMICAL TECHNIQUES 89-99 7.0 Introduction 7.1 Objectives 7.2 Histochemical Techniques: An Introduction 7.2.1 Lipids Staining 7.2.2 Protein and Amino Acids 7.2.3 PAS Reaction (Periodic Acid Schiff) 7.2.4 Enzyme 7.3 Answers to Check Your Progress Questions 7.4 Summary 7.5 Key Words 7.6 Self Assessment Questions and Exercises 7.7 Further Readings UNIT 8 MICRO SLIDE PREPARATION 100-110 8.0 Introduction 8.1 Objectives 8.2 Micro Slide Preparation 8.2.1 Preparation Techniques: Dry Mounts, Wet Mount, Squash, Staining 8.2.2 Whole Mounts 8.3 Answers to Check Your Progress Questions 8.4 Summary 8.5 Key Words 8.6 Self Assessment Questions and Exercices 8.7 Further Readings UNIT 9 SECTIONING OF BIOLOGICAL SPECIMENS 111-123 9.0 Introduction 9.1 Objectives 9.2 Sectioning of Biological Specimens: Basic Concepts 9.2.1 Free Hand Sectioning 9.2.2 Hand Microtome 9.2.3 Rotary Microtome Sectioning 9.2.4 Sledge Microtome 9.3 Embedding Methods 9.4 Answers to Check Your Progress Questions 9.5 Summary 9.6 Key Words 9.7 Self Assessment Questions and Exercises 9.8 Further Readings UNIT 10 DEWAXING