2013 Small World In Motion Competition

Budding yeast with Gal1 promoter driving unstable GFP and myo1-mcherry during induction. Upon switch from glucose to galactose media, GFP is activated stochastically.

Lu Bai

Affiliation
Penn State University
University Park, Pennsylvania, USA

Anyone who’s enjoyed a glass of wine or fresh-baked bread can appreciate the amazing scientific properties of yeast. Yet for scientists such as Lu Bai, yeast is capable of so much more. Bai works with yeast to study gene expression, thanks to how much one can change its genetic content. In the world of genetic research, yeast allows means of experimentation that is much harder with human cells, fruit flies, and other samples.

Bai is Assistant Professor of Biochemistry and Molecular Biology and Assistant Professor of Physics at Pennsylvania State University. She is also one of the Honorable Mention winners of the Nikon Small World in Motion competition for her captivating video, “Budding Yeast.” In her video, we see how this amazing species shows genes expressing themselves in reproducing cells.

Aesthetically, the video is mesmerizing. In it, we see the yeast doubling and growing in size. It becomes hypnotic to watch as it takes on a new form with every new addition. But take a closer look, and like everything in the Nikon Small World universe, there is more than meets the eye.

Though the video is less than 30 seconds in length, what we’re actually viewing is a movie that took place over the course of 10 hours. Every time we see the yeast double (which happens every second of the video or so), we’re actually seeing is about 1.5 hours of real-time work. There is more to this technique than just speeding up the video, however. It’s hard to keep the environment for yeast to be “happy,” as Bai puts it. One has to maintain the environment for growth, while keeping the focus constant and the sample stage stable so the video comes out clearly.

Even more important is the green fluorescence we see in Bai’s video, highlighting the genes she is interested in studying. “All the cells you see here are from the same family: they originate from one ‘mother cell’ and grow from there, so genetically they are all the same,” Bai says. “At the beginning of the movie, we changed the yeast growth conditions, and the cells respond by ‘turn-on’ some genes, and one of them has green fluorescence. In other words, the green color in the cells represent their response to changing environment. If you look carefully, you will notice that different cells respond at very different time, some after two hours, and some after seven hours. In other words, the cells have ‘individuality’ and make different ‘decisions.’ This is the basis of both my research and fascination!”

Bai’s research is important, because in the human body, the same type of cells may also have different gene expressions, and it may sometimes cause disease - like cancer - for example. Understanding how and why individuality occurs, then, is the key to unlocking how to prevent them from occurring in the first place, if needed. To do this, her work involves characterizing the differences among cells, understanding if the individuality that occurs is good or bad, and trying to work out how the cells control these differences. Bai’s research is part of a larger body of work being done that hopes to uncover why these phenomenon take place. It can only be done studying cells one-by-one like we see here.

In Bai’s video, we see the work to solve one of modern science’s current mysteries. Her contribution to Nikon Small World in Motion showcases the best of the competition - a passion for science, shown through incredible research, and in a way that is aesthetically pleasing to the eye and that the general public can start to understand.

Bai says that while the scientific community started by studying yeast, they are working toward addressing similar questions in higher species. Simply put she says, “We are trying to understand the mechanism so that one day we can learn to control it.”

An egg mass of the spruce budworm, Choristoneura occidentalis, hatching over a 2-hour time period. The egg mass is affixed to a needle of a Douglas fir tree

Dr. Ward B. Strong

Affiliation
BC Ministry of Forests, Lands and Natural Resource Operations
Vernon, British Columbia, Canada
Technique
Stereomicroscopy with time-lapse photography
Magnification
15x

To the naked eye, there’s nothing exciting or even noticeable about the tiny egg mass on a spruce needle. But look a little closer, at just the right time, and you might just see something fascinating.

Dr. Ward Strong of Vernon, Canada happened to be in the right place at the right time, looking at a specific spruce needle. The result? An enthralling stop-motion video featuring fifty spruce budworms hatching simultaneously from an egg mass, chewing their way out as individual larvae. Through use of a Greenough Stereo Microscope at 15x magnification paired with a Nikon D90 controlled with a computer to take pictures every five seconds, Dr. Strong was able to capture about 1400 images, which he then assembled into a movie at twenty frames per second. “What I think is amazing is when you’re looking at these by eye, in real time, it happens very slowly, over two hours. It’s an inconspicuous egg mass on a spruce needle; you wouldn’t notice it,” says Dr. Strong. “But speed it up, you see how truly fascinating it is. Simultaneously it froths and bubbles, as each budworm begins squirming and wiggling to escape.”

Spruce budworms lay their eggs in summer and hatch in August, producing only a single generation each year. Once they escape the egg mass, each tiny larva burrows into a needle and spends the winter there, safe from natural enemies and the elements. In spring, it resumes feeding on the tree. It’s these feeding habits that caught the attention of Dr. Strong and his research team, who for the past three years have studied the genomics of pest resistance in trees in collaboration with University of British Columbia in Vancouver.

Thanks to increasingly rapid climate change, trees are becoming less and less adapted to the places they live – and therefore more susceptible to pests and insects such as spruce budworms.  While some trees are more resistant to pests than others, Dr. Strong’s team noted that the current breeding system is too slow to keep up with the changing climate.  They set out to assist this process with genomic marker assisted selection – a brand-new mechanism they hope will someday improve breeding in trees that are resistant to pests.  Spruce budworms such as those captured in Dr. Strong’s winning entry have been vital to this study, as researchers observed which trees the budworms prefer to feed upon, and which trees are resistant.  Identifying markers in the tree DNA that correlate with weevil resistance will ultimately speed the process of breeding weevil-resistant trees, thus improving our ability to respond to climate change. As a model system for a new technology, this project also promises applications in breeding for any desirable trait in trees.

According to Dr. Strong, this winning video captures more than just the research behind it.  “As a scientist with a passion for the arts, photography through microscopes has been a very strong theme in my life and career,” says Dr. Strong.  “I’m enthralled with the opportunity provided by this technology, to show the world what is happening under the microscope. There’s an astounding, beautiful, fascinating world under there… if we only take a moment to look a little closer.”

Ophryoglena atra (ciliates) feasting on a copepod larva

Wim van Egmond

Affiliation
Micropolitan Museum
Berkel en Rodenrijs, Zuid-Holland, The Netherlands
Technique
Differential Interference Contrast
Magnification
160x

This video plays out more like a microscopic thriller, with two ciliates scavenging inside the body of a dead organism. Like vultures, ciliates detect dying animals on which to feed. In this case the “victim” is the larvae of a copopod – although how they entered the body is a mystery to van Egmond, who was surprised to capture the pair on their “great escape” once they finished feeding.

Capturing such a scene took a combination of patience and luck, as van Egmond carefully laid out his slides, examining them every so often. He first took shots focusing on the ciliates feeding on the organic material. Thirty minutes passed before he returned to examine the slides, and just managed to catch them fleeing the scene. “It’s strange and beautiful, these simple organisms have no eyes, no ears, no organs,” says van Egmond. “But still, they sense one another. One escapes, and the other follows in an instant. It’s fascinating.”