Meeting on Sat, June 1st., 2013
Earthworm Dissection Notes (29 KB)
The topic for this session was “Bacteria”. Lewis gave a short introductory talk and this was followed by practical work.
Microtomy (1.8 MB)
Notes on Wax Embedding-2 (28 KB)
Wax embedding protocol (1.2 MB)
‘Live’ yoghurt was used as a source of harmless bacteria for staining and mounting. Several stains were available for experimentation and the Gram staining technique was introduced for all to try. This work provided a useful opportunity for participants to learn (or sharpen up) technique for using oil-immersion systems.
The notes below are:
- A summary of the points made in the introductory talk,
- A protocol for Gram staining of bacterial specimens.
Nineteen members attended the Group’s May meeting. Lewis made a short presentation of background information about Earthworms to provide the context for a practical session during which a general dissection was performed by each of those present. The following notes and ‘photo.s give a flavour of what happened at the session. There were plenty of favourable comments about this meeting and all those present were keen to express their appreciation of the work done by Keith and Maureen in the provision of an excellent lunch and refreshments throughout the day.
ICENI MICROSCOPY STUDY GROUP
Four of our sessions during 2012 are being devoted to the cutting, processing and mounting of thin sections of biological materials; these are being led by Lewis Woolnough. The first session covered free-hand cutting of “easy” plant material and, in the second, sections were cut with hand- and bench-microtomes. Some very nicely stained sections were mounted and all participants, some of whom had never done this sort of work previously, said that they had enjoyed the experience.
The diagram and notes that follow provide guidance for anyone who may wish to apply this procedure for themselves.
Honeybee Dissection – The Head
Meeting report for 5/Nov/2011
This is an account of the IMSG’s meeting of 5th November 2011. The dissections were introduced by Peter Sunderland whose introduction appears below; these included a handout from Snodgrass’s Anatomy of the Honey Bee (1956). A “what we did” guide follows on page 2 for others who may wish to make these dissections themselves.
In order to spot the glands you must know what the glands do, if we dissect a bee at random without knowing its age it could be a difficult dissection without the following information. The glands in the bees head change throughout their life, most beekeepers know the hypopharyngeal glands produces royal jelly and protein identical with food given to the larvae. After a period, three to four weeks, they change their function and start to shrink; they then produce the enzymes used in converting nectar into honey, but there is a sting in the tail, should the colony go queenless, the hypopharyngeal glands can be re-activated to produce royal jelly and brood food. The enzymes are added to liquid food during ingestion by the bee. The hypopharyngeal glands are the largest in the head and it is split into two branches and can contain up to 500 nodules per string, the nodules individually are called acini.
The next one to interest us in dissection is the post cerebal glands, the ones in the head are the salivary branch (and one in the thorax are called the thoracic branch; of no interest to us today). The salivary gland is predominantly on the underside of the head – see page 52 in your hand-out. The salivary glands are under pressure and they are held back by the salivary syringe on page 53, marked SLV. The ones we shall be dissecting today is the obviously the top half of the drawing, on the right-hand side of the drawing (page 53 D) shows the unique shape, a bit like an ivy bud, as opposed to the hypopharyngeal which is completely round.
The next gland of interest is the mandibular gland and it is interesting to read in Snodgrass on page 58 (see handout) the statement “the function the mandibular gland is not definitely known”. The lumen of the gland (which the actual tube) acts as a reservoir. The secretion is under pressure and flows out when released. The outlet is lens-shaped, between two stiff curved plates, one of which closes the outlet by springing to reverse curvature when released. The outlet opens when this plate is pulled by a ligament connecting it to the hypopharyngeal plate, which is pulled by a muscle attached to the tentorial bridge. The secretion contains much 10-hydroxydec-2-enoic acid in workers, and 9-oxodec-2-enoic acid in queens. The former acid is the principal fatty component of larval food. In foraging workers the secretion also contains a scent, mainly 2-heptanone, which seems to be an alerting pheromone. Some experiments have been claimed to show that mandibular gland secretion is used in working wax and propolis, but no 10-hydroxydec-2-enoic acid has been revealed by analyses of wax from combs.
The acid in the queens’ mandibular glands can attract drones from at least 10 feet (3m), and enables workers in a travelling swarm to know that they have a queen with them, but it has only a weak power to inhibit colonies from rearing queens. Queen’s mandibular glands also contain pheromones which enable workers to recognise queens when they are close enough to touch them.
The mandibular glands are shown on page 53 H, very easy to confuse with the hypopharygeal glands – but the hypopharygeal are not in bag (shape and size is everything).
ince the glands have a thickness and should not be crushed, a cavity slide in required. Prepare some cavity slides by ringing with gold size; to give sufficient depth, several rings on top of each other will be needed; make the diameter that of your cover slips; we used 16 mm. Allow to dry.
Fresh bees are needed; they are best brought alive and killed just before dissection using ethyl acetate. Cut off the heads of about six using a scalpel; this gives a cleaner result than pulling.
Warm a microscope slide in a spirit lamp flame and rub some beeswax onto it. When some of the wax melts, remove the slide from the flame and put onto the molten wax five heads the right way up (to access the hypophrangeal and mandibular glands) and one face down (for the head, or post cerebal, salivary glands). We were warned not to use too much wax but it is soft so, if there is a little too much, it is easy to cut through the excess.
Using a scalpel, cut along the top of the head and then down the sides through the eyes until the frons, the piece of exo-skeleton freed, can be peeled downwards towards the tongue.
The hypophrangeal and mandibular glands are now visible and can be teased out using seekers or needles. In early November, many bees were found to have a layer of fat lying between the exo-skeleton and the hypophrangeal glands. Manipulation is simpler if a drop of water is put onto the head. With careful manipulation and some luck, a string of hypophrangeal acini can be removed – only a length is wanted, not a mass.
It was interesting that bees varied. Some hypophrangeal glands were an opaque pale pink but others a more amber colour and almost translucent. Some had a layer of fat between the exo-skeleton and the hypophrangeal glands but others did not. After referring to Peter it seems that the younger bees producing brood food would have opaque pale pink acini but older bees using their hypophrangeal glands to produce sucrase (or invertase) would have amber translucent acini and their muscles for discharging the brood food would have atrophied to become a part of the fat body.
Each mandibular gland is attached to a mandible and this will have to be snipped off its hinge.
In a similar way, using the head secured face down, a cut may be made into one of the rearward bulges at the back of the head and a section of one of the head salivary glands removed.
After removal, the length is put into the slide cavity and a drop of lacto-phenol added. This should be ready stained with Trypan Blue. A further small drop of water is also added to ensure that the cavity is full. The outer edge of the cover slip is then painted with size, turned upside down and gently pressed onto the slide ring of size – this forces some water out but the water does not prevent the rings sealing together. The edge of the cover slip should be further ringed for completeness but this can be done later.
A report on the visit to the Centre for Environment, Fisheries & Aquaculture Science (CEFAS) at Lowestoft by members of the Iceni Microscopy Study Group on 24th November 2010
The main focus of the visit (originally) was to see how CEFAS made sections of fish, the purpose was to try and find out CEFAS’s techniques and methods for doing this type of work with a view to replicating some of the ideas and methods within the Iceni. We had for the first part of the visit a laboratory tour by Lorraine Greenwood, head of the laboratory, who showed us the various microtomes they used and also the equipment which professionally sharpens the microtome blades. Lorraine talked us through the processes of how they used a form of resin to encapsulate their specimens and mounted them onto wooden blocks before they took off the sections, this process is quite different from the Iceni’s way of doing things where we use paraffin wax which is much, much softer. We were shown a selection of slides dating back to the 1960s showing sections through fish eggs, by studying the eggs and how the eggs were forming they were able to establish the egg/breeding status of the fish at that time. Eggs were either laid or reabsorbed when breeding/environmental conditions were not ideal. Using a computer they were able to count the number of viable eggs and then build charts and graphs and forecast whether fishing should be increased or decreased.
The second part of the day entailed a visit to the otolith laboratory. Fish have bones in their heads called otoliths (oto’ meaning ear and lith’ meaning stone). These bones help the fish to keeping its balance in the water. When an otolith is removed from a fish, sectioned into thin slices and viewed through a microscope, it reveals a pattern of light and dark concentric rings. We were shown around by the head of this department, Glenn Saunders, who was ably supported by Suzy Baldry. We were shown how they mount the otoliths (which can be as big as your little finger or as big as your thumbnail) onto to metal moulds which were then filled with black resin to form a cake which was then fitted into a computer driven modified grinding machine which sliced it into very thin slices, the blade was diamond impregnated and was a little bit less than 1mm thick. These thin slices were then laid onto a glass slide, approximately 50mm x 110mm and a clear cover glass of the same size (the biggest I have ever seen) and interestingly set down by using Bondit resin, the same clear resin used for making paperweights. We were offered the opportunity to look at some slices through a microscope and you could indeed see the rings rather like tree rings which determine the age of the fish, it was most interesting to see the procedures. We also asked how they kept a record of all these slides and 100% traceability, they said they had slides going back to 1945 and they showed us a map which was marked in decreasing squares, the smallest being a kilometre, each slide had a number to identify where it came from within the world. On should bear in mind that they take samples from all around them world and they do work for other countries, as well as Britain and the EU. Some of the specimens had odd location codes, where fishermen had been economic with the truth, with codes denoting that the fish were caught in “Peterborough”.
The consensus was that everyone enjoyed the visit and that it would be good to go back again, CEFAS in turn have said that we may pay a return visit. The Iceni has sent thank you letters to all three of CEFAS’s staff that played a part during our visit.”
Iceni Microscopy Study Group, Meeting on Saturday, Oct. 30th. at Barsham.
Attendance: David Blackwood, John Blakesy, Peter Clarke, Bob Curtis, David Galliford, John Hunt, Brian Norman, Peter Sunderland and Lewis Woolnough.
Avis kindly gave up her day to organise the domestic arrangements for us. This included the provision of a splendid lunch, including a welcome contribution from Mike in the form of soup. Those present were very grateful and wish to record their sincere thanks to Avis and Mike.
The morning was devoted to the subject of Phase Contrast. Lewis gave an illustrated talk about the basic theory behind the creation of this effect to achieve enhanced contrast between the specimen and its background. A demonstration of the technique for setting up a microscope for viewing with phase contrast followed and this led into a session of practical work for those present; working in groups of two or three, they enthusiastically set about applying their understanding and skills to great effect.
After lunch, our Chairman, Peter Sunderland, led a discussion about the planning of future activities in general and a possible visit to the Fish Research Lab’s in Lowestoft, in particular. There was a useful exchange of opinions leading to approval of what Peter had in mind. Peter had arranged to make a preliminary visit to Lowestoft on 1-11-10 and it was hoped that a visit for Iceni members might be arranged before the end of 2010.
Peter Clarke then set up a splendid puzzle. He provided three samples of sand; one was from a recent volcanic eruption in Iceland, the second was river sand, again from Iceland, and the third some beach sand from the Suffolk coast. Our challenge was to identify each one correctly. Peter then revealed the answers and pointed out features that provided the key clues for sorting out the samples. It transpired that the group had done remarkably well on this task and had had a thoroughly enjoyable time doing it. I am sure everyone would wish to join me in thanking Peter for organising this exercise.
The day concluded with a brief and informal introductory look at some microtomes. Lewis showed a Hand-, Bench- and Cambridge Rocking Microtome. Some other people had acquired the last-mentioned type, of varying ages, and these were looked at as well. The preparation of wax-embedded specimens and the actual cutting of sections is a subject we plan to address in much more detail soon.
The feedback following this meeting suggests that things went well on the day and that those present had had an enjoyable and useful experience.
Iceni Microscopy Study Group
‘Basic Microscopy For Beginners’ course
First session, Saturday 6th March
There were seven applicants for this course. Tthey started arriving at 9.15am, and were soon talking to each other and drinking coffee or tea. Lewis Woolnough got the session under way at 10.00am, with a ‘bang’, (Keith Wilkinson burst a balloon on the word ‘GO’). From then on the morning unfolded with information on light,images and basic microscopy with most people taking notes (handouts were available). This was followed with practical involvement with candles, lenses and cards with participants writing down what happened as the position of the lighted candle was changed. The image of the flame becoming bigger or smaller and clearer or not so clear, as it was cast onto a screen by the convex lens. The lenses were changed with different results, but all arriving at a satisfactory conclusion.
Everyone enjoyed the usual lunch, after which Lewis demonstrated how to make pipettes from glass tubing and various types of ‘droppers’from glass rods, all attending trying their hand. The afternoon passed very quickly, with all present having had a successful day, when the session came to an end at a little after 4.00pm.
Report of meeting held on Saturday 13th February
There was a good attendance, with eight potential members, who had been invited to meet the members of the Group. Seven of these were interested in the ‘Basic Microscopy for beginners’ course, starting on 6th March and running for three sessions.(20th march and 10th April). The other visitor was Jeremy Quinlan of SBKA, and a BBKA Master Beekeeper, who has expressed an interest in joining the ICENI Group. The day started with a brief Committe meeting, following the usual format, at the end of which Peter Sunderland, chairman gave a detailed report on his thoughts of what the group could do this year. These ideas were welcomed by all, although it will not be possible to achieve them all in one year. The meeting came to an end with a discussion on what advice should be given to beekeepers at the Nosema day, on which there was divided opinion. The secretary suggested that if SBKA were to put out their information, that it should make sure that it be made clear that it was information produced by SBKA and not the ICENI Microscopy Group. 27/02/10
Report of Iceni meeting, held on Saturday 23rd January
All but one of the original members were present, in addition to which another seven potential members joined us for introductory sessions.
The President of the Waveney Beekeepers Group, Michael Venn, having been invited, arrived mid morning and was introduced to the Group. Mike was most intrigued with the work and enthusiasm of the Group, and sent a very complimentary letter to the secretary some days later. He was introduced to all present, individually, and reminisced with Karin and David from Hampshire, (his old ‘stamping ground’). After the usual lunch, Mike took his leave of the Group, fully pleased with his visit.
One of the potential members decided to join the Group and five of the others soon signed up for the ‘Basic Microscopy’ course, commencing on 6th March. Once again, a very productive day.
Report of Iceni meeting held on 31st October 2009
Location: Grange farm Centre, Barsham, Beccles.
This was THE DAY, assessment day, for the BBKA Microscopy Exam.
The members arrived from 9.15am and were offered beverages and biscuits on arrival, as usual, but today with encouragement also. The assessors began to arrive at 9.45am and after a welcome ‘cuppa’, outlined the plan of action. Each member took up their allotted area and the assessors got to work, asking questions,viewing slides etc, etc, etc. As the morning wore on the atmosphere became more relaxed, and by 1.20pm everthing was finished. Lunch was served, as usual, all members and assessors sitting round the table, chatting about topical affairs.
Ideas for the future of the group were discussed and the assessors remarked on how impressed they were with the ‘set-up’ and that we were lucky to have such a good place in which to practise microscopy, albeit a little off the beaten track.
After lunch the BBKA assessors went into the other room, to discuss their results, emerging about fifty minutes later. Obviously they could not give any information as to how things had gone, at this stage, but said that we would be notified soon. When e.mail addresses had been exchanged, people began to disappear, some having a distance to travel. The ‘victims’, (members taking the assessment) agreed that it had not been such a bad experience and were quite confident of the outcome.
We shall see!
Report of meeting held Saturday 17th October
The meeting opened at 10.00am, with six members present. Lewis Woolnough, President, started by congratulating Peter Sunderland on his success at the National Exhibition, where he had submitted slides and received an award. WELL DONE Peter!
The group then held an informal discussion, regarding specific items, in connection with the forthcoming BBKA Microscopy Assessment,on 31st October. What would the format be, time of starting, what was permissible as regards literature for reference? (ABSOLUTELY NON ALLOWED), etc,etc,etc.
Q. Were Nosema slides to be made up from a sample of 30 bees?
A. It is required to make up a slide from your own sample of bees and to say whether it is infected or NOT. If positive you have to count the numbers and classify the infection as Low, Medium or High as appropriate.
Q. What was the next step if the sample was clear?
A. Make another slide for confirmation.
The question of the ‘Bailey Comb Exchange’ system came up, with Peter explaining what was entailed. Lewis then presented a ‘check’ list of items, which he considered would be required for the members to take for the assessment, and this was discussed at some length. The composition of a pollen grain was explained, and how many bees would be required.
Peter Sunderland distributed Price Lists from TCS Biosciences Ltd, Dyes and Stains. Lewis then reminded members of the proposed visit to The Natural History Museum, London, on 14th November, when Celia Davies will be presenting a lecture concerning Bees. Possibly 6/8 members would be going and it was suggested that we meet at Stowmarket station, to all go on the same train. Lewis,Keith and Peter to discuss and finalise.
The next meeting, the assessment, will be held on 31st October and our visit to The Quekett Club will be the last one of this session.
Arrangements for next years’ meetings are still to be agreed.
Report of Iceni meeting, held on Saturday 3rd October
Another successful meeting was held, at Esmerelda’s, Grange farm Centre, Barsham, Beccles.
The business of the day commenced at 10.15am, with all present taking part in theoretical and practical revision,for the forthcoming BBKA assessment. A light lunch was enjoyed at around 12.30pm, followed by a short Committee meeting.
The Chairman, Peter Sunderland handed round copies of a paper on Nosema, which he had produced for use at the SBKA nosema Day. It was discussed by the members and a few additions made, which Keith Wilkinson, Administrator agreed to re-produce with alterations.
The Treasurer then gave a report of the state of the finances and reminded members that subscriptions for the last quarter were now due. He then went on to give a report of the meeting held on 5th September, with Jeremy Quinlan,SBKA representative, all members agreeing with the arrangements made.
Keith mentioned courses and work for 2010, suggesting that a business meeting be held towards the end of November,to discuss and plan for next year. After a little discussion it was agreed that Lewis Woolnough, President, Peter Sunderland, Chairman and Keith Wilkinson, Administrator should meet to discuss and plan this, as they had done previously.
The afternoon was then given over to more practical microscopy. The meeting closed at 4.30pm, as usual.
Report of IMSG meeting, held on Saturday 5th September
The meeting commenced at 10.15am, with the introduction of Jeremy Quinlan, of Suffolk Beekeepers association, who had joined the meeting to discuss the organisation of a ‘Nosema day’, to be held in Suffolk, at which members of the IMSG will assist with technical input. Members then got down to the usual routine of practise preparation for the BBKA assessment, on October 31st, at Barsham. Lewis, Peter, Jeremy and Keith,adjourned to the other room, to discuss the ‘Nosema Day’ arrangements. This meeting lasted for just over an hour, and the agenda covered, venue, date and time, organiser, work for the day,volunteers, advertising, assessment of inspection and report of findings and other relevant details. It was a very Satisfactory meeting, details of which are on the IMSG file. After lunch, served by Avis, Muriel recovering from a spell of hospitalisation, the members got down to Microscopy again. It was an enjoyable day, with exchanges of information and knowledge between all present, and the meeting finished at 4.30pm.
REPORT OF IMSG MEETING 15th August
The meeting was held at Esmerelda’s, Grange Farm, Barsham, Beccles and commenced at 10.15am, with a short committee meeting, (bi-monthly). Peter Sunderland reminded everyone that today the lst of the pollens should be completed, after which Keith Wilkinson reported on the financial situation.The Group finances are in good order, with no excessive payments outstanding. Then followed a short discussion on future meetings, and points regarding the BBKA Microscopy examination were discussed and noted for later consideration.The Meeting closed at 11.15am. A full Report of the Meeting is electronically filed, along with others.
Lewis remarked he had observed, that part of the procedure for setting up Compound Microscopes didn’t appear to always be followed, and many members asked him to go through it again, with them. The outcome was success, and satisfactory to all present.
The members then got down to the main object of the meting, MICROSCOPY PRACTISE.
A light lunch was enjoyed at 12.45pm, after the members were persuaded to leave their work for a short period. It is most difficult to drag them away from their microscopes, even for Food & Drink!
The members began to drift away, reluctantly, at around 4.30pm, some of them with a long drive ahead of them.
Iceni meeting Saturday 25th July 2009
The Iceni microscopy Study Group held another successful meeting on Saturday 25th July, in preparation for the forthcoming BBKA Microscopy Assessment in October. The morning session opened with a talk by Don Cooper,of WNKLBA, on bee diseases. As past SBI in Norfolk, Don had plenty to refresh our memories about, plus some things we may not even have known. The proposed discussion, ” Plan of Action- advice to beekeepers”was briefly mentioned but postponed to a later date.
A paper by Peter Clarke, on ‘The Theory of Sedimentation and Centrifugation’ was available for perusal by members during the day, but it will be more fully considered, on a future occasion.
After the usual light lunch,to which members have become accustomed, the practical session began with great enthusiasm. The meeting finished
at around 4.45pm.
Report of Iceni meeting Saturday 4th July 2009
The ICENI microscopy Group, once again held a very successful meeting on Saturday 4th July, at Esmerelda’s, Grange Farm Centre, Barsham, Beccles.
The Administrator opened the meeting at 10.30am, with details of all notices and correspondence received since last meeting. High on the agenda were details of an ‘AD’ to be placed in the Ipswich & E.Suffolk BKA August Newsletter, and a date to see Jeremy Quinlan,of I&ESBKA, for
discussions on a ‘Nosema Day’, proposed for early next year. The BBKA Assessment Day has been fixed for Saturday 31st October, when 4 Assessors from BBKA will descend on ‘Esmerelda’s’ for this. Seven members of Iceni will be taking the assessment.
In view of a clash with a WBG function on Saturday 1st August, our next meeting was brought forward to 25th July. Further meetings have been arranged up to the end of October.
After a light ‘ploughman’s style’ lunch, the members got down to practical sessions, mostly dissection of bees, and Don Cooper demonstrated extracting pollen grains by sedimentation, by ‘Centrifuge’.
Report of Iceni Monthly Meeting, held Saturday 6th June, at Grange Farm Centre.
The proceedings opened at 10.30am,with a committee meeting, which went on until lunchtime. Items discussed are recorded in the report of that meeting, held by the administrator.
The main interest was the setting up of the ‘website’, and it was agreed that Peter Clarke would continue to be the ‘Webmaster’, with articles from members being sent to the administrator , to be converted to a form which Peter could upload. It was also decided that the website address should appear on all letter headings, etc.
The members then enjoyed a ploughman’s type lunch, before getting down to practical revision.
The next meeting will be held on Saturday 4th July, at the grange farm Centre.
Origins of the group
The Iceni Microscopy Study Group began life as ‘Waveney Microscopy Study Group’, on Saturday 12th July 2008, when a group of 14 people gathered at Easton College, Norwich to commence the first session of a microscopy course, which would run on eight Saturdays, ending on 13th December with assessments.
It was the ‘brain child’ of Peter Sunderland, a Waveney beekeeper and keen microscopist, who felt that beekeepers could do something for themselves, at the same time improving bee health, locally, and maybe educate others in bee diseases in the area, particularly as bees and bee keepers were going through a difficult time.
Peter therefore made contact with microscopists, bee experts, lecturers, National Bee Unit, Central Science laboratory,bee inspectors and others, in order to plan and organise the course, which would run on eight Saturdays, at Easton College, up to 13th December.
Members of the group also met at Esmerelda’s, Grange Farm Centre, every two weeks during September, October and November,to gain practise and further expertise of what they had learned at Easton College, principally the use of microscopes to examine their bees for bee diseases such as acarine, nosema, EFB and AFB, which all require access to a microscope.
The object being to help other beekeepers if at all possible.Pollen recognition and slide making was also a major actvity, being connected with bees.
Such was the enthusiasm and commitment,when the course ended, that they decided to continue their activities on a monthly basis, and thus re-formed, on 7th March 2009, as ‘Iceni Microscopy Study Group’, under ‘Club’ rules.They then decided to extend their activities to cover ‘Microscopy in general’, and now welcome new members who are interested in microscopy covering other aspects of study,not only bees.
The strength of the Club is in the enthusiasm and commitment,some members having long journeys to the meetings at Esmerelda’s, Grange Farm Centre, Hall Road, Barsham, Beccles.