Cutting Edge Techniques in Biophysics Biochemistry and Cell Biology: From Principle to Applications Anupam Jyoti, Neetu Mishra.

Format:

Book

Author/Creator:

Jyoti, Anupam, Author.

Contributor:

Jyoti, Anupam, editor.

Mishra, Neetu, editor.

Language:

English

Subjects (All):

Biophysics.

Physical Description:

1 online resource (153 pages)

Edition:

1st ed.

Place of Publication:

Singapore : Bentham Science Publishers, [2019]

Summary:

Advances in biomedical research have had a profound effect on human health outcomes over the last century. Biophysical, biochemical and cellular techniques are now the backbone of modern biomedical research. Understanding these laboratory techniques is a prerequisite for investigating the processes responsible for human diseases and discovering new treatment methods. Cutting Edge Techniques in Biophysics, Biochemistry and Cell Biology: From Principle to Applications Provides information about basic and advanced analytical techniques applied in specific areas of life science and biomedical research. Key Features: - Book chapters present a broad overview of sophisticated analytical techniques used in biophysics, biochemistry and cell biology. - Techniques covered include in vitro cell culture techniques, flow cytometry, real time PCR, X-ray crystallography, RNA sequencing - Information about industrial and biomedical applications of techniques, (drug screening, disease models, functional assays, disease diagnosis, gene expression analysis and protein structure determination) is included. The book is an excellent introduction for students (as a textbook) and researchers (as a reference work). The information it presents will prepare readers to understand and develop research methods in life science laboratories for different projects and activities.

Contents:

Cover

Title

Copyright

End User License Agreement

Contents

Foreword

Preface

List of Contributors

Animal Cell Culture: From Fundamental Techniques to Biomedical Applications

Neetu Mishra* and Joyita Banerjee

1. INTRODUCTION

2. FUNDAMENTAL REQUIREMENTS IN ANIMAL CELL CULTURE LABORATORY

3. TYPES OF CELLS

4. TYPES OF CELL CULTURES

4.1. Primary Cell Culture

4.2. Secondary Cell Culture

4.3. Maintenance of Cell Cultures or Sub-culturing of Cells

4.3.1. Adherent Cell Cultures

4.3.2. Suspension Cell Cultures

5. CRYOPRESERVATION OR FREEZING OF CELLS

6. THAWING OF FROZEN CELLS

7. CELL COUNTING

8. CELL VIABILITY AND CELL PROLIFERATION ASSAY BY USING MTT

9. CELL CULTURE CONTAMINANTS

9.1. Overview of Biological Contaminants in Cell Culture

9.1.1. Bacterial Contamination

9.1.2. Mycoplasma Contamination

9.1.3. Mold Contamination

9.1.4. Yeast Contamination

9.1.5. Virus Contamination

9.2. Cross Contamination

10. APPLICATIONS OF ANIMAL CELL CULTURE IN BIOMEDICAL RESEARCH

CONCLUSION

CONSENT FOR PUBLICATION

CONFLICT OF INTEREST

ACKNOWLEDGEMENTS

REFERENCES

Real Time PCR as a Diagnostic Tool in Biomedical Sciences

Ritismita Devi1, Rohit Saluja2,3 and Swapnil Sinha1,*

INTRODUCTION

Detection Chemistries

Intercalating Dyes

Fluorescent Labeled Oligonucleotide Probes

TaqMan Probes

Molecular Beacons Probe

Nucleic Acid Analogues

APPLICATIONS

Differential Gene Expression

Cancer Diagnostics

Bacterial Infection Diagnosis

Food Adulteration

Mutational Analysis for Predicting Predisposition to Disease(s)

Advanced Techniques

CONCLUDING REMARKS

Flow Cytometry: Basics and Applications

Juhi Saxena1 and Anupam Jyoti2,*

INTRODUCTION.

Components of a Flow Cytometer

Fluidics

Optics

Electronics

Fluorochromes

Maximal Excitation and Maximal Emission

Fluorescence Compensation

Data Analysis

Gates

1. Apoptosis Detection

Procedure: Apoptosis Detection Using Annexin-PI Labelling [7].

2. Cell Cycle Analysis

Procedure: DNA Cell Cycle Analysis [9].

3. Immunophenotyping

Procedure: Identification of Human Neutrophils in Blood [11].

4. Free Radical Generation

Procedure: Detection of Free Radicals in Human Neutrophils [13].

5. Cell Sorting

Procedure: Sorting of CD4+ and CD8+ Cells from WBCs [15].

Protein Structure Determination Using Experimental Phasing Technique

Vijay Kumar Srivastava*

EXPERIMENTAL PHASING METHODS USING HEAVY ATOMS (HAS)

REPLACEMENT OF SULFUR ATOMS WITH SELENIUM

STRUCTURE SOLUTION

Determining Heavy Atom Positions

Refinements of Heavy Atom Parameters

Phase Determination

Software Used for Structure Solution through MIR

Structure Refinement and Validation

Different Programs to Refine the Macromolecular Structures, are as follows:

REFMAC5

Phenix Refinement

Fourier Maps and Map Interpretation

Omit Maps

Geometric Analysis

Current Advancements in Macromolecular X-ray Crystallography

ACKNOWLEDGMENTS

Essentials of Recombinant Protein Production

Satyajeet Das and Sanket Kaushik*

Types of Host Cells Used for Heterologous Protein Production

Bacterial Expression System

Fungi as Expression System

Animal Based Expression Systems

Importance of Vectors in Heterologous Gene Expression

Purification of Recombinant Protein.

Applications of Heterologous Protein Production

In Silico Modeling and Drug Designing

Gauri Misra* and Neetu Jabalia

PRINCIPLE

Homology Modelling

Molecular Docking

Virtual High-Throughput Screening

Quantitative Structure-Activity Relationship (QSAR)

Conformational Analysis

Monte-Carlo Simulation

Molecular Dynamic (MD) Simulations

Generation of Chemical Structures

Molecular Structure Visualization

Determination of Molecular Properties

ADMET Properties

APPLICATIONS OF MOLECULAR MODELING

Determination of Mechanism of Action of Drugs

Identification of Novel Drug Targets

Determination of Biological Activities of Unknown Compounds

Design of Novel Drugs

ADVANCED TECHNIQUES

Multi-Target Drugs Discovery

Site-Identification by Ligand Competitive Saturation

Biomolecular Crystallography and Its Applications

Nagendra Singh*

Protein Production

Crystallization

X-ray Scattering

Molecular Replacement

X-ray Crystallography in Medicine

X-ray Crystallography in Genetic Diseases

X-ray Crystallography in Infectious Diseases

X-ray Crystallography in Drug Metabolism

X-ray Crystallography in Vaccine Development

RNA Sequencing Technology for Biomedical Sciences

Sandeep Ameta* and Roberta Menafra

PRINCIPLE AND TECHNOLOGY

a. Pre-Sequencing:

b. Sequencing:

c. Post-Sequencing:

QUALITY CONTROL STRATEGIES IN THE NEXT-GENERATION SEQUENCING

ADVANCED TECHNIQUES.

Single Cell RNA Sequencing

Direct RNA Sequencing

Immunoelectrophoresis: Recent Advances and Applications

Vinod Singh Gour* and Ravneet Chug

Principle

Applications

a. Pathogen/Protein Detection

b. Cancer Diagnosis

c. Protease Activity

d. Disorders

e. Differentiation in Protein Profile

Limitations

Advantages

CROSS IMMUNOELECTROPHORESIS

Limitation

ROCKET IMMUNOELECTROPHORESIS (RIE)

IMMUNOELECTROPHORESIS 2D

Subject Index

Back Cover.

Notes:

Description based on print version record.

ISBN:

9789811422867

9811422869

OCLC:

1432628625