Microscopy for beekeepers
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This document provides an overview of microscopy for beekeepers. It discusses what microscopy is and the main types of microscopes, including light microscopes, electron microscopes, and scanning probe microscopes. It focuses on setting up and using light microscopes to examine pollen, bee biology, and bee parasites. Specific topics covered include how to prepare slides of pollen from flowers and honey, examining bee anatomy like legs and wings, and looking at bee parasites like tracheal mites. The document recommends areas to examine under the microscope and references for further information.
Microscopy for beekeepers
- 1. Microscopy for Beekeepers Chris Cardew 31st January 2015
- 2. Content • What is microscopy? • Types of microscope – How to set up the light microscope. – How to make slides • What to look at – Pollen – Bee biology – Bee parasites • Where next
- 3. What is Microscopy • Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye • There are three well-known branches of microscopy: optical (light), electron, and scanning probe microscopy.
- 4. Types of microscope • Optical or Light Microscopy – Dissecting x40 times magnification – Compound up to x 1000 times • Electron Beam – Scanning electron microscope • Up to x 250,000 – Transmission electron microscopy • Up to x 1,000 higher magnification than light
- 5. Dissecting Microscope • Low magnification observation of a sample. • Using light reflected from the surface of an object rather than transmitted through it. • Produces a three- dimensional visualization of the sample being examined
- 6. Compound Microscope • Viewing small cells, or thin sections of organs or tissues placed on a glass mounting slide. • The specimens are thin enough that light can pass through them from below. • Magnifications of compound microscopes are generally range from 40x to 1000x • One objective lens (the lens above the specimen) used at a time
- 7. The x1000 objective lens must be used with oil to prevent distortion and aberration. The oil (yellow) has similar refractive index to Crown glass Glass slide http://goo.gl/tre3AQ The parts of the microscope.
- 8. Setting up a microscope
- 9. Setting up for a larger audience Camera Microscope Laptop TV/Projector USB HDMI Heliconsoft 3D software Eyepiece with graticule
- 10. What to look at and why? • Pollen – Monitor the flowers that bees visit – Identify honey – Make reference slides and photographs • Bee biology • Compare to other insects e.g wasp, bumblebee, solitary bees • Bee parasites – Tracheal mite, Varroa
- 11. Pollen
- 12. Identifying Pollen - flower • Pollen from the flower anther – Pick the flower with anther – Add gelatine to a slide, brush the anther onto the gelatine. – (Take a blob of gelatine on some tweezers and brush it over the flower anther then place the blob onto a slide) – Place a cover slip over the pollen. – Place the slide on a heating plate – The gelatine will spread out under the cover slip
- 13. Mallow from the flower x400
- 14. Mallow 100um diameter (0.1mm) Pollen drawings by Dorothy Hodges Forget me not one of the smallest pollen grains One micrometre (1 um) is 1/1000th of a millimetre (1mm) One millimetre 10 mallow pollen grains
- 15. Hazel pollen (26th Jan 2015)
- 16. Identifying pollen - Bee • Pollen taken into the hive by the honeybees as observed by Dorothy Hodges • The honeybee mixes honey and/or nectar with the pollen with the result that the colour can vary • Careful observation is required to to be sure of the source of the pollen • Better to look at the pollen under a microscope • Pollen is mixed with water and then placed onto a slide
- 17. Identifying pollen- Honey • Extracting pollen from honey • centrifuge and sedimentation. • Take 10g of honey (centre of the jar) • Make slide as for flower • Pollen concentrations • These change by pollen type • Pollen coefficients were introduced to help with honey identification (more detail later) • Eva Crane – A book of Honey • Other methods of identifying honey include: – Conduction – Refractive index. • Mellissopalynology • The study of pollen contained in honey
- 18. Heather x400 What pollen is this? A sample of Heather Honey
- 19. Heather (x1000)
- 21. Honey Identification • Rex Sawyer method: – Take 10g of honey – Mix with 250g of water – Either place in centrifuge or sediment out – Place a drop of the sediment on a slide – Pollen count must be in the region of 200 – Calculate % of pollen types – Using pollen coefficient calculate % honey Taken from ‘Honey Identification by Rex Sawyer’
- 22. Honey Identification Plant source Pollen % PC Pollen coefficient (1000 grains per 10g honey) Relative Quantities of honey Pollen/PC Percentage composition of honey Rubus 36 50 0.72 22.2 Castenea 26 1000 0.026 0.8 Tilia (Lime) 21 10 2.1 64.7 Trifolium 9 50 0.18 5.5 Ligustrum 3 25 0.12 3.7 Others 5 50 0.1 3.1 TOTALS 100 - 3.246 100 This was a honey sample claimed as Lime Honey and justified by the above calculation Taken from ‘Honey Identification by Rex Sawyer’
- 23. Rosaceae family: Hawthorn, Apple, Plum, Blackthorn, Wild Cherry Raspberry, Blackberry, Pear, Mountain Ash All have very similar looking pollen grains Pollen grain drawings by Dorothy Hodges
- 24. Using calibrated graticule accurate measurements can be taken
- 25. Honey identification • Summary – Identify the pollen grains in the honey sample – Calculate the % pollen in the sample – Calculate the % honey using the pollen coefficient – Use technology to accurately measure the pollen grains to help with identification. – Make reference slides directly from the flowers to compare with the pollen in the honey
- 26. Purple Loosestrife is unique (in UK) Three different length stamens mean that is has three different sized pollen grains
- 28. Gathering nuts in May Pollen grains are the same size to a bee as nuts , small fruit and seeds are to humans
- 29. Bee Biology
- 30. Bee biology • Bee dissection – the dissecting microscope • Looking at bee legs • Looking at bee wings • Comparing the bee to other insects
- 31. Chemicals for slide preparation • Numount • Final mounting of bee parts onto a slide • Glycerine Jelly • Mounting of pollen and other very small parts onto a slide • Sodium Hydroxide • Softening of bee parts for slide mounting e.g. leg • Acetic Acid • Decalcification of bee parts for slide mounting • Isopropyl Alcohol (IPA) • Preservation (fixing) of bee parts • Toluene • Dehydrating and clearing of bee parts (process with IPA)
- 35. Hind leg x 400 Pollen Press
- 39. Fore leg x400 Antenna Cleaner
- 40. Honeybee worker tongue x 100
- 41. Tongue x400
- 42. Antenna x100
- 43. Antenna x400
- 44. Two different focus points of the antenna (x600). Using 3D software will enhance the final image
- 45. Bee Parasites
- 46. Bee disease and parasites • Viruses (for example Sacbrood) cannot be seen with a light microscope. • Bacteria can be seen at magnifications of x1000. – However there are only two physical shapes that bacteria can have, either ‘sausage’ shape - bacillus, or ‘ball’ shaped coccus. – This applies to disease causing bacteria as well as the many normal strains, therefore using shape alone the strain of bacteria cannot be identified. – This is really the area of professional laboratories • http://www.brunelmicroscopes.co.uk/apiculturebeeke eping.html
- 47. Female tracheal mites enter the first thoracic spiracle of the young honeybee
- 48. Why bother? • Many reasons but one of which might be to pursue the beekeeping modules, WBKA, BBKA The Modules are written examinations held at a centre in your region with each paper taking 1½hrs. You can take up to 4 modules in each session. There are seven modules to be studied: Module 1 - Honey bee Management Module 2 - Honey bee Products and Forage Module 3 - Honey bee Pests, Diseases and Poisoning Module 5 - Honey bee Biology Module 6 - Honey bee Behaviour Module 7 - Selection & Breeding of Honey bees Module 8 - Honey bee Management, Health and History Microscopy
- 49. Where next • Series of seminars in the autumn. – Microscopy – Bee disease – Bee biology – Husbandry – Product presentation – ??? • Interest groups; – pollen – bee biology – looking at wasps and bumblebees
- 50. References • Honey Identification • Rex Sawyer. Northern Books • Pollen Loads of the Honey Bee • Dorothy Hodges • Anatomy and Dissection of the Honeybee • Harry Arthur Dade • Bee • Rose Lynn Fisher. Princeton Architectural Press • A Book of Honey • Eva Crane
- 51. Any Questions?