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Barnard Awards for Videomicrographs – 2020
Sinclair Stammers judged the videomicrographs before they were revealed at the virtual Annual Exhibition of Microscopy on Wednesday 30th September 2020, and had the difficult job of deciding which of the excellent videos from members of the Quekett, the Iceni Microscopy Study Group and the Postal Microscopical Society were of a sufficiently high standard to deserve a certificate.
The entries that deserved certificates were announced after the lecture on Saturday 3rd October. Sinclair decided that certificates should be awarded to David Linstead (Collotheca unfurls its sails), Willem Cramer (Carp louse) and Daryan Chitsaz (Confocal time-lapse).
Chris Algar
Rotifers feeding
The rotifers were collected from a bird bath. Filmed using an Olympus BH-2 microscope with bright field illumination and 10× and 20× objectives. Canon EOS 7D camera. Edited using iMovie software.
Sinclair’s comments: Excellent clear video of the rotifer Philodina feeding on algal cells, bright field works well.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Vorticella
The Vorticella were collected from a garden pond and photographed using various objective magnifications on an Olympus BH-2 microscope with bright field illumination. Canon EOS 7D camera. Edited using iMovie software.
Sinclair’s comments: Vorticella is a familiar subject but the rapid contractions of the wavy myoneme fibres at the core of the stalk are quite mesmerising, good stable video of a fascinating subject.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Chris Thomas
Wood from an angiosperm tree
Sinclair’s comments: Thinking in 3 dimensions can be a hard concept to get across to students, and this video very successfully and succinctly demonstrates the 3 image planes that timber scientists use routinely to characterise wood in Angiosperms.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Wood from a gymnosperm tree
Sinclair’s comments: Similarly this video demonstrates very well the 3 image planes used to describe wood in Gymnosperms.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Daryan Chitsaz – Barnard Award
Sinclair’s comments: Wonderful Time Lapse! Utterly magical image, it’s great the way confocal microscopy show such delicate structures.
Macrophage munching on myelin
This video displays 2 macrophages (left panel, large cells at the top and bottom) interacting with human myelin debris (floating particles in bright magenta / white). On the right is a zoomed-in image of the boxed area, highlighting a myelin particle being phagocytosed. In diseases like multiple sclerosis, myelin (a substance coating neurons which is essential for their function) is mis-identified by the immune system as foreign, and cells such as these invade the brain to dispose of it. Here, macrophages isolated from human donor blood were treated with Mitotracker Green, a dye which specifically labels mitochondria. Myelin isolated from human tissue was added to the culture moments before imaging, and was labelled with the lipophilic dye Fluoromyelin Red (pseudocoloured in magenta), which also labelled the highly dynamic membranes of the macrophages.
Microscopy: 3D image stack time series were acquired for 120 15-second intervals on a Zeiss LSM880 with Airyscan, using a 63× 1.4 NA oil-immersion objective. This confocal system utilizes a honeycomb-like detector array rather than a single detector, which improves sensitivity and allows multiple lines of pixels to rapidly be acquired simultaneously – both features ideal for 3D live cell imaging.
Photography: Raw Airyscan images were processed using Zen software (deconvolving each detector’s image and summing them), and 2D maximum projections of the image stacks were exported. A rolling-ball background subtraction was used for the magenta channel, and contrast of both channels were locally enhanced using the CLAHE algorithm in Fiji (an ImageJ build). The scalebar is 10 µm.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Three oligodendrocytes on wires
These videos were from experiments studying oligodendrocytes (OLs), specialized brain cells which wrap up neurons analogously to insulation around wires. OLs can be cultured on plastic transparent fibers which they ensheath like neurons, allowing their growth dynamics to be easily imaged in a controlled environment. The cells are labelled by infecting them with engineered viruses carrying genes for fluorescent proteins. A caveat is that OLs often grow both on the fibers, as well as on the culture dish base below them, as is displayed here. These videos were depth-coded such that cool colours correspond to cellular processes at the base of the plate, while hotter colours are on the fibers above, with cell bodies appearing white. The fibers themselves are not labelled. Other, more dynamic cell types can be seen in the culture, including a domineering astrocyte at the top.
Microscopy: 3D image stack time series were acquired for 10 5-minute intervals on a Zeiss LSM880 with Airyscan, using a 20µ 0.8 NA objective. This confocal system utilizes a honeycomb-like detector array rather than a single detector, which improves sensitivity and allows multiple lines of pixels to rapidly be acquired simultaneously.
Photography: Raw Airyscan images were processed using Zen software (deconvolving each detector’s image, normalizing, and summing them). Images were depth coded with a custom macro and contrast was locally enhanced using the CLAHE algorithm in Fiji (an ImageJ build). 3 scenes were put side by side to create the montage.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Withering brain cells
Here, 4 glial cells were continuously imaged after environmental controls were switched off (required for the cells to survive). The lipophilic dye Nile Red was used, which emits different coloured light when it stains cell membranes (shown in magenta) or certain types of vesicles (green / white). The dye was improperly dissolved, resulting in grey shards in the culture media. Two stoic oligodendrocytes can be seen in the center with a multitude of spindly process. On the bottom and right are two astrocytes, which are rapidly withdrawing their processes, disrupting the oligodendrocytes.
Microscopy: Timelapse images were acquired for 15 2-minute intervals on a Zeiss LSM880 with Airyscan. This confocal system utilizes a honeycomb-like detector array rather than a single detector, which improves sensitivity and allows multiple lines of pixels to rapidly be acquired simultaneously. A 565 nm laser was used with low pass or high pass filters was used for the green and magenta channels, respectively.
Photography: Raw Airyscan images were processed using Zen software (deconvolving each detector’s image and summing them). Contrast was linearly adjusted.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
David Linstead
Collotheca unfurls its sails – Barnard Award
Zeiss Standard microscope with DIC, ordinary halogen illumination and a Reichert fluorite 16× objective. Panasonic Lumix GX9 camera coupled afocally with 30mm Sigma micro4/3 prime lens and a Leitz Periplan 10×20 eyepiece (with 28mm eyecup thread). Processing of the raw video was with Serif Movie Plus X6.
Sinclair’s comments: Exquisite unfurling of the infundibulum in the rotifer Collotheca which opens like a blossom, they have extremely long tentacle-like cilia surrounding the corona. Differential Interference Contrast shows the fine tentacles beautifully.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Floscularia
Zeiss Standard microscope with DIC, ordinary halogen illuminationand a Reichert fluorite 16× objective. Canon M3 camera coupled afocally with 30mm Sigma micro4/3 prime lens and a Leitz Periplan 10×20 eyepiece (with 28mm eyecup thread). Processing of the raw video was with Serif Movie Plus X6.
Sinclair’s comments: Very nice composition and clarity of focus, although the fuzzy area bottom right is a tad distracting.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Stephanoceros plays pinball with Chlamydomonas
Zeiss Standard microscope with DIC, ordinary halogen illumination and a Reichert fluorite 16× objective. Panasonic Lumix GX9 camera coupled afocally with 30mm Sigma micro4/3 prime lens and a Leitz Periplan 10×20 eyepiece (with 28mm eyecup thread). Processing of the raw video was with Serif Movie Plus X6.
Sinclair’s comments: Beautiful image! perfect composition, fine detail and smooth movements are superb.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Deborah Kapell
Two rotifers, two amoebas and an air bubble
These were taken from a broad puddle that exists for a few days after heavy rain then dries up until the next rain. Bdelloid rotifers are the perfect inhabitants of puddles – they contract into cysts when the environment dries then come out again when the water returns. The sample was taken in April, 2020 from a recurring puddle in Prospect Park, Brooklyn, NY, USA.
Taken using an Olympus BH-2 BHTU, 20× objective, phase contrast. Canon EOS 5D Mark II camera. Color was enhanced in PhotoShop.
Sinclair’s comments: Very elegant clear footage, phase contrast works very well with these microorganisms.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Grenham Ireland
Breakfast for worm larva
A spionid worm larva was isolated using a glass pipette from a marine plankton sample and confined in a glass ring with copepod eggs to try and replicate observed feeding behaviour.
Bright field image using 4× objective and Olympus BH-2 microscope of sample in a 2 mm glass ring held in a plastic dish. Recorded with a Panasonic GH6 camera and edited with Movie maker.
Sinclair’s comments: Very amusing Voice Over, I like the way the video tells a story about the behaviour of this tiny marine annelid. I hope it didn’t miss breakfast! However I would criticise the rather messy composition, with no attempt to find a neutral white balance!
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Sinclair Stammers
Fairy shrimps
Chirocephalus diaphanus from Hergest Ridge shallow ponds
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Willem Cramer
Carp louse – Barnard Award
This highly flattened fish parasite is covered with a back shield. At the bottom of the head, the first lower jaws are transformed into suction cups with which they attach to fish. The chest piece consists of only four segments, each of which carries a pair of split-leg shaped swim legs. The abdomen is little developed and not externally articulated.
Argulus falls under the Branchiura class. They belong to the crustaceans. At least 50 species of Argulus are known. This native (NL) carp louse (Argulus foliaceus) is a common pest on Koi fish, for example. The name carp louse is somewhat misleading, as it can also be found on many other fish, including brackish water fish. However, the preference is for cyprinids, including the goldfish.
With the help of the bushy legs, the carp louse is able to swim from one fish to another, in addition, it can move over the fish body. The animal is visible to the naked eye and can grow to about 8 mm in size. Argulus has a mean sting. It is located between the eyes and can kill a small fish in one sting. The carp louse attaches to the fish with two suction cups and many other extremities and pierces the epidermis of the fish with the sting. Poison is then injected and blood is drawn up.
The wounds left behind are difficult to heal and can be a source of secondary conditions such as secondary inflammation, bacterial infections, fungal growth, possible transmission of viruses by Argulus. Loss of skin function with massive infection.
Argulus is difficult to combat. On the one hand, this is because the animal can survive without a host for up to three weeks. On the other hand, because the eggs are resistant to many pesticides.
Motic SMZ-171 stereomicroscope, darkfield, Moticam ProS5 Plus camera
Sinclair’s comments: Fab quality dark field video microscopy. My sympathy goes with the fish that have to suffer these awful ectoparasites.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Skeletal muscle tissue
Mammals have three types of muscle tissue:
- Skeletal muscle tissue that consists of parallel-arranged multinucleated, striated muscle cells with a fast, powerful will-dependent contraction.
- Heart tissue that consists of mononuclear, striated branched cells, the contraction of which is synchronous, rhythmic, powerful and autonomous.
- Smooth muscle cells are spindle-shaped and show no transverse banding; their contraction is slow and not subject to will.
The striated skeletal muscle cells or muscle fibres are long, cylindrical, multinucleated giant cells. The length of the individual muscle fibres varies in humans between 1 mm (as in the tympanic cavity) up to 30 cm (in the sartorius muscle) while the diameter is between 10 and 100 microns. Inside a muscle, the diameter of the fibres is fairly constant.
In the images the transverse banding of skeletal muscle fibres can be seen, which matches the banding of the neighbouring muscle fibres. The lateral- and cross-sections also clearly show the cell nuclei with their contents such as the nucleolus and chromatin.
Tympanic cavity = a small cavity surrounding the bones of the middle ear.
Sartorius muscle = a long, thin, superficial muscle that runs down the length of the thigh.
Panthera U with Plan UC 40× and 100× objectives, Moticam S12 camera
Sinclair’s comments: Super quality stained thin sections of muscle cells, the track and zoom used in this video is very effective and helps direct the viewer to the interesting features.
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
Volvox bloom
Volvox is a popular alga among microscopists because of the beautiful images it produces under the microscope. The
Volvox in the video was unexpectedly found in a pool in Eindhoven, the Netherlands in early November 2019. There were so many of these species present that one could speak of a bloom.
The green alga
Volvox is a colony of cells that have started to work together. Some cells catch the light, others provide movement or reproduction. They have become so dependent on each other that you can speak of a multicellular organism.
Some “colonies” are no more than a collection of cells that do exactly the same, but the spherical
Volvox is different. Cells work together there and specialize. All cells are on the outside of the sphere. There are cells with whip hairs or cilia for movement and cells that provide for reproduction.
In good times, germ cells immediately grow into daughter colonies that float inside the mother globe. They continue independently as the parent colony opens and then dies. As the winter approaches, sex comes around the corner. The germ cell changes into a packet of sperm cells, or enlarges to an egg cell. Sperm cells are released and swim to the eggs to fertilize them. The fertilized egg forms a hard, thick wall.
Volvox can survive winter well in this capsule.
With thanks to Harry Nouwen, Royal Dutch Natural History Association.
Motic BA310E with Plan Apo 4× to 40× objectives, Moticam 1080 camera
Sinclair’s comments: Very nice clean images, however I have a few criticisms. The coenobia all look like they have been filmed under laboratory conditions, in other words flat in one plane. Volvox look so much more beautiful when they are swimming freely in 3 dimensions!
Click the arrow to start the video; click the symbol to the left of “vimeo” for a larger version
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