Table of Contents

Title Page

Copyright

List of Figures

List of Tables

Acknowledgements

Introduction

Health and Safety

Chapter 1: Measurements and Calculations

1.1 Units and measurements

1.2 Measuring the volumes of liquids

1.3 Pipetting

1.4 Weighing

1.5 Calculations

Chapter 2: Preparing Solutions

2.1 Common terms defining solutions

2.2 Precautions in making solutions

2.3 Making solutions

2.4 Dilutions to prepare standard solutions

2.5 Molar solutions

2.6 Calculations involving solutions

Chapter 3: Separation of Liquids and Solids

3.1 Filtration

3.2 Centrifugation

3.3 Chromatography

3.4 Electrophoresis

Chapter 4: Common Techniques and Equipment

4.1 Titration

4.2 Spectrophotometry

4.3 Aseptic techniques

4.4 Disinfectants

Chapter 5: Microscopy and Histology

5.1 Light microscopy

5.2 Slide preparation

5.3 Cell Counting

Chapter 6: Cardiorespiratory Measurements

6.1 Techniques to investigate cardiovascular function

6.2 Techniques to investigate respiratory function

Chapter 7: Recording and Presenting Data

7.1 Keeping a laboratory book

7.2 Presentation of data

7.3 Recording data in tables

7.4 Presenting data in graphs

7.5 Describing data statistically

Recommended Reading

Appendix 1: Rules for Powers

Appendix 2: Rules for Logarithms

Appendix 3: Factors to Consider When Making Solutions

Appendix 4: Principle of Spectrophotometry

Appendix 5: Descriptive Statistics and Formulae

Appendix 6: Using Software to Draw Tables, Graphs and Calculating Descriptive Statistics

Index

List of Figures

Chapter 1  Measurements and Calculations

Figure 1.1 Accuracy and precision shown on a target
Figure 1.2a Beaker
Figure 1.2b Conical (Erlenmeyer) flask
Figure 1.2c Measuring cylinder
Figure 1.2d Burette
Figure 1.2e Volumetric flask
Figure 1.2f Pipette
Figure 1.3 Location of the meniscus of a liquid
Figure 1.4 Parts of a Gilson pipette
Figure 1.5 Circle of pipettes
Figure 1.6 Tips used with pipettes
Figure 1.7 Setting the volume on a Gilson pipette
Figure 1.8 Types of pipette tips
Figure 1.9 Pipette inserted into beaker
Figure 1.10 Ejecting tip using tip ejector button
Figure 1.11 Multichannel pipette
Figure 1.12 Pasteur pipette
Figure 1.13 Portable digital top-loading balance
Figure 1.14 Plastic boat
Figure 1.15 Spatula
Figure 1.16 A typical analytical balance

 

Chapter 2  Preparing Solutions

Figure 2.1 Wash bottle
Figure 2.2 Serial dilution of a stock solution

 

Chapter 3  Separation of Liquids and Solids

Figure 3.1a Funnel
Figure 3.1b Cone-shaped filter paper
Figure 3.1c Fluted-shaped filter paper
Figure 3.2 Buchner funnel
Figure 3.3 Side-arm filtering flask
Figure 3.4a Low-speed centrifuge
Figure 3.4b High-speed centrifuge
Figure 3.5 Microhaematocrit centrifuge
Figure 3.6a Capillary tube
Figure 3.6b Capillary tube showing separated components after centrifugation
Figure 3.7 Chromatogram showing baseline, spots and solvent front
Figure 3.8 Gel electrophoresis chamber
Figure 3.9 Gel electrophoresis wells

 

Chapter 4  Common Techniques and Equipment

Figure 4.1 pH paper
Figure 4.2 pH meter
Figure 4.3 Spectrophotometer
Figure 4.4 Cuvettes
Figure 4.5 Standard curve of light absorption against concentration
Figure 4.6 Inoculating loop
Figure 4.7 Bunsen burner
Figure 4.8 Petri dish
Figure 4.9a Streak plate
Figure 4.9b Plate with colonies

 

Chapter 5  Microscopy and Histology

Figure 5.1 Binocular light microscope
Figure 5.2a Objective lens of microscope
Figure 5.2b Ocular lens of microscope (eyepieces)
Figure 5.2c Stage and clips to hold slide
Figure 5.3 Adjustment of ocular lens
Figure 5.4 Ocular micrometer
Figure 5.5 Stage micrometer
Figure 5.6 Overlaying of ocular and stage micrometers
Figure 5.7 Unstained slide with wax covered section
Figure 5.8a H & E staining tray
Figure 5.8b Placing slide in Erlich's haematoxylin
Figure 5.8c Making a permanent mount of stained tissue
Figure 5.8d Permanent slide (stained)
Figure 5.9 Haemocytometer
Figure 5.10 Haemocytometer grid with dimensions

 

Chapter 6  Cardiorespiratory Measurements

Figure 6.1a Sphygmomanometer
Figure 6.1b Sphygmomanometer and stethoscope
Figure 6.2 Automatic blood pressure monitor
Figure 6.3 Finger prick pen
Figure 6.4a Lancet
Figure 6.4b Pushing lancet into pen
Figure 6.4c Removing base of lancet
Figure 6.4d Pen containing inserted lancet
Figure 6.4e Pen placed perpendicular to side of finger
Figure 6.4f Ejection of used lancet from pen
Figure 6.5 Wet seal spirometer
Figure 6.6a Controls of wet seal spirometer
Figure 6.6b Controls to switch between spirometer and atmosphere
Figure 6.7 Loading vitalograph chart paper onto moveable tray
Figure 6.8a Vitalograph at the beginning of a blow
Figure 6.8b Vitalograph at the end of a blow
Figure 6.9 Electronic spirometer (MicroPlus)

 

Chapter 7  Recording and Presenting Data

Figure 7.1 Example of line graph
Figure 7.2 Example of bar graph
Figure 7.3 Example of scatter graph
Figure 7.4 Drawing line of best fit using linear regression
Figure 7.5 Testing the line of best fit
Figure 7.6 Pie chart

 

Appendix 4  Principle of Spectrophotometry

Figure A4.1 Components of a spectrophotometer

 

Appendix 6  Using Software to Draw Tables, Graphs and Calculating Descriptive Statistics

Figure A6.1 Bar graph showing changes in the mean heart rate (±SEM) of students at rest and during a maths test

List of Tables

Chapter 1  Measurements and Calculations

Table 1.1 Common measurements and their units and symbols
Table 1.2 Common prefixes
Table 1.3 Common measurement units of volume
Table 1.4 Volume ranges for Gilson pipettes

 

Chapter 2  Preparing Solutions

Table 2.1 Standard solutions prepared from a stock solution by dilution

 

Chapter 4  Common Techniques and Equipment

Table 4.1 Acids and bases commonly used in the laboratory
Table 4.2 Volumes of stock and water used to produce diluted standard solutions
Table 4.3 Absorbances of the standard and unknown solutions

 

Chapter 7  Recording and Presenting Data

Table 7.1 Examples of independent and dependent variables
Table 7.2 Glucose tolerance test for normal and diabetic persons

 

Appendix 6  Using Software to Draw Tables, Graphs and Calculating Descriptive Statistics

Table A6.1 Example of a table using Microsoft Word or Excel
Table A6.2 Descriptive statistics

Acknowledgements

We are extremely grateful to Varoopah Senthuran for taking the majority of photographs, and her enthusiasm, support and assistance in a variety of other tasks during the production of this book. Discussions on book contents with John Allum and Alberto Sanchez-Medina at the initial stage of writing the book were highly appreciated by the authors. We are also grateful to Stefano Casalotti and Winston Morgan for their suggestions and helpful feedback on the manuscript. We would also like to thank the following, for their help in providing the various apparatus in different laboratories, and for suggestions in taking better photographs: Susan Harrison, Raymond Stoker, Stephen Garrad, Duncan Kenedy, Kevin Clough, Manchu Ambihaiphan, Ashford Clovis, and Keith Eley.

We would also like to thank the publishing editors of Wiley Publishers for their patience, understanding and guidance in the preparation of this book.

Introduction

‘Biosciences’ is a broad term covering a wide range of subjects, including biology, biochemistry, biomedical science, biotechnology, forensic science, microbiology, physiology, pharmacology, and toxicology. Studying the biosciences involves a large amount of practical work in laboratories. These sessions involve following instructions from the lab instructor, following a practical schedule, learning techniques, taking measurements, observing and recording data, calculating and presenting data.

You will find that in performing practical laboratory tasks, techniques are often repeated (e.g. pipetting volumes, preparing solutions or producing calibration curves). You will be required to apply these fundamental skills across the bioscience subject areas.

This book is intended to act as a laboratory guide to help you to complete these practical tasks successfully by focusing on:

The book is intended for those who are new to, or have little experience of the laboratory techniques and equipment used in studying biosciences. It may also be useful to those who need to refresh some of their knowledge of laboratory skills, or those doing projects at both undergraduate and postgraduate levels. This book emphasizes ‘essential’ skills and the practical steps required to use equipment and learn techniques. The authors assume that the reader has very little prior knowledge of techniques in the biosciences. To make the process easier, we have emphasized the step-by-step approach in the practical procedures, together with a wide range of photographs of equipment and accessories.

This is not intended to be a comprehensive text book, so you will find that the background theory to the techniques (Chapters 3, 4, 5 and 6) is brief. Also, some techniques in the various bioscience fields, and some particularly advanced techniques that you may be using in years two and three of your degree may not have been included. Please note that the steps shown for procedures are not universal, and you will come across some differences in practice, depending on the technique and equipment you are using.

Chapter 1 covers the fundamental basis of measurements and units, and common mathematical calculations you may come across. You must understand terms which are commonly used, such as ‘moles’, ‘molarity’ and ‘dilutions’. In addition, you must be able to convert between common prefixes or powers of ten. You will have to practise doing calculations involving moles, molarity and powers. Pictures of glassware commonly used in the lab are included.

Chapter 2 gives details of common terms associated with solutions and how different types of solutions are prepared.

Chapter 3 contains details of techniques used to separate liquids and solids, such as filtration, centrifugation, chromatography and gel electrophoresis. Some of the chromatography methods using expensive equipment will not be routinely used in undergraduate bioscience practicals, especially at entry level. Therefore, instead of detailed procedures of the methods, we give the basic principles and an outline of the methods. It is intended to help distinguish the methods which may appear confusing for a beginner.

Chapter 4 contains details of a wider variety of techniques and equipment, including spectrophotometry, pH meters, titration and aseptic techniques.

Chapter 5 contains details on light microscopy, histology, and cell counting.

Chapter 6 covers a range of techniques, investigating cardiopulmonary function in humans.

Chapter 7, the final chapter, describes how to record and present your data, with examples of typical tables and graphs.

Appendices in bold face to highlight the section give more details of laws of powers and logarithms, more theory on some techniques (e.g. spectrophotometry), and the use of common software such as Microsoft Word and Excel to produce tables, graphs and descriptive statistics. There is only a brief description of statistical analysis (descriptive statistics), so you will need to refer to statistical texts to investigate differences in mean data (inferential statistics).

We hope this book will be a useful ‘companion’ to the lab schedule and lecturers' instructions, and will help to make the student more independent in the laboratory.

Mohammed Meah and Elizabeth Kebede-Westhead

University of East London

Health and Safety

Before performing any laboratory work, it is essential that health and safety considerations are followed with regard to hazards and risk to you and others. Make sure you follow the basic important rules in a laboratory as shown below.

Chapter 1

Measurements and Calculations

Learning outcomes

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