In vitro quantitation of Theileria parva sporozoites for use - TropMed ...

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Appendices 153 _______________________________________________________________________________________________ Add 150 µl medium in each well using a multichannel pipette (Use a fresh set of tips for each column). Pipette-mix about 3 times at each step. Label the plate Transfer to CO 2 incubator at 37°C (record time) Leave undisturbed for 10 days Day 10 Destroy fungi contaminated cells with 10N NaOH. Check cell cultures for clumps Make cyto-centrifuge smears: mount slides with a filter card, centrifuge 50 to 100 µl at 215 g for 5 min Stain with 12.5% Giemsa’s stain in Sörensen's phosphate buffer (0.008 M, pH 7.2) Score cultures for the presence or the absence of schizonts
154 Appendices ______________________________________________________________________________________________ APPENDIX II Titration curves of Theileria parva stabilates extracted by homogeniser and manually (as depicted in Figure 7 of chapter 3) Predicted proportion of positive wells wells 1 0.5 0 Homogeniser -4 -3.5 -3 -2.5 -2 -1.5 Manual -4 -3.5 -3 -2.5 -2 -1.5 Stabilate dose (ln t.e.)
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Victor Mbao In vitro quantitation o
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II Front cover photograph Copyright
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V ACKNOWLEDGEMENTS This work was ma
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VII TABLE OF CONTENTS ACKNOWLEDGEME
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IX APPENDIX I .....................
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General Introduction 13 ___________
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General Introduction 15 ___________
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Chapter 1 - Quantitation of Theiler
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20 Chapter 1: Quantitation of Theil
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Chapter 2: Objectives 43 __________
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Chapter 3 -Comparison of two Theile
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48 Chapter 3: Theileria parva extra
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Chapter 4: Suspension media and mul
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Chapter 5 - Snap freezing of Theile
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70 Chapter 5: Snap freezing of Thei
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Chapter 6: Storage of Theileria par
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Chapter 7: Lyophilisation of Theile
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Page 104 and 105: Chapter 8: SNA and refinement of ti
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Page 120 and 121: Chapter 9: General Discussion 119 _
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Page 130 and 131: References 129 ____________________
Page 148: APPENDICES
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Page 166: SUMMARY
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Page 178: Curriculum Vitae
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Page 184 and 185: List of Abbreviations 183 _________
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