Plant Anatomy An Applied Approach (Lecture ppt) Ch10_PracticalMicrotechnique

Plant Anatomy An Applied Approach (Lecture ppt) Ch10_PracticalMicrotechnique

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Unformatted text preview: Ch.10 Ch.10 Practical microtechnique Practical Gwo­Ing Liao Department of Life Sciences, National Cheng Kung University Safety considerations Safety Health & Safety Wear appropriate protective Clothing Glove Goggle Disposable active carbon face mask p.170 p.170 http://www.medicalexamglove.com/laborato ry_glove_vinyl.jpg http://www.dryeyepain.com/Goggles.htm Safety considerations Safety p.170 Appropriate use of fume cupboard At least two persons Waste disposal 國國國國國國國國國國國 http://www.eose.ncku.edu.tw/~guotf/lab/machin e/P1010350.jpg 國國國國國國國國國國國國國國國國國 http://www.iosh.gov.tw/Publish.aspx?cnid=25 國 國 國 國 國 國 國 國 國 國 國 國 國 國 (GHS)- 國 國國國國國國國國國國國國國國國國國國國國 國國國國 http://ghs.cla.gov.tw/CHT/intro/search.aspx?cssid=3 國國國國國國國國國國國國國 國 國國國國國國國國 國 國 GHS 國 國 國 國 p.170 Materials Materials “Standard” plant → study “Standard plant list” → teaching Selected study material (Appendix 1, p.195) (Appendix Anatomical characters Families Local floras Anatomical leaf, stem, a few examples (p.195~200) (p.195~200) softwood, hardwood (p.201~202) (p.201~202) Materials Materials p.170 Typical & Atypical Zea maize ←→ monocotyledon Typical plant- morphology, physiology … Zea maize, barley, rice…… peas, lettuce, sunflower, tobacco, beet, tomato, potato, Arabidopsis thaliana…… Materials- Anatomical studies Materials- p.171 Fresh material & Dried herbarium material Cell structure and histology - better to fix Fixative - killing and preserving cell contents Kill the protoplast without distortion Preserve to fix the fine detail Harden the material Methods- Fixatives Methods- p.171~173 Fixatives Alcohol and formaldehyde-based fixatives FAA, FPA→ wax wax formalin 國 國 國 -acetic acid - alcohol國 Table formalin acetic alcohol Table 10.1 國 formalin -propionic acid -alcohol 10.1 Non-coagulating fixatives osmium tetroxide 國 國 國 國 , acrolein 國 國 國 tetroxide acrolein glutaraldehyde 國 國 國 , formaldehyde→ plastics Methods- Dehydration Dehydration → Infiltration p.173~175 Dehydration→ Infiltration Alcohol series→ xylene 國 國 國 → xylene wax wax TBA series→ wax 國 Table 10.2 國 wax • TBA 25.5℃ • Wax 50~70℃ 國 48hrs Methods- Sectioning Methods- p.175 Sectioning- magnifications of 100~400x Freehand sections Rotary microtome wax-embedded 10μm specimens thick http://www.meyerinst.com/html/leica/rm2135.htm http://www.nmnh.si.edu/rtp/students/2006/images/botany_day52.jpg Methods- Sectioning Methods- p.175~179 Sliding microtome 15~30μm thick Wood 國 Fig. 10.1 國 Twigs 國 Fig. 10.2~3 國 Leaves 國 Fig. 10.4 國 silica bodies- 10% hydrofluoric acid 50% alcohol Methods- Clearing MethodsClearing 50% alcohol ↓ water ↓ sodium hypochlorite p.177~179 Methods- Staining Methods- p.179~183 Staining temporary & permanent Temporary stains Double stain FABIL Safranin and haematoxylin Safranin and fast green 11.Basic histochemistry p.237, CD-ROM Factfile index Methods Methods Preparing permanent slides Freehand sections 國 Table 10.3 國 Paraffin sections p.183~187 Methods Methods Preparation of surfaces Leaf surfaces 國 Fig. 10.6 國 Stem surfaces Surface replicas Cuticular preparations Clearing material p.189~191 Methods Methods Standard levels 國 Fig. 10.7 國 Leaf Petioples Stems Roots p.191 Microscopy Microscopy p.191~194 LM light microscopy (OM optical microscopy) TEM transmission electron microscopy SM stereo microscopy SEM scanning electron microscopy Microscopy and the interpretation…… Appendices- methods in microscopy p.205, CD-ROM The virtual plant ...
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