sábado, 27 de mayo de 2017

Economical Fluorescence for Stereo Microscopy: Speeds Workflow and Brightens Education

Economical Fluorescence for Stereo Microscopy: Speeds Workflow and Brightens Education


Economical Fluorescence for Stereo Microscopy: Speeds Workflow and Brightens Education


Fluorescence is a critical tool for investigating biological phenomena, but until recently, has been limited primarily to higher-end compound microscopes for observing cellular phenomena. Over the past 10 years, fluorescence has extended to stereo microscopes, driven largely by interest in genetics, genomics, and neuroscience that required imaging whole organisms for key animal models, including fruit flies (Drosophila melanogaster), worms (C. elegans), zebrafish (Dania rerio), and frogs (Xenopus sp.), and plant models, especially Arabidopsis thaliana. The instrumentation has been expensive, often restricting availability to just one high end fluorescence stereo microscope in a lab or department.
NIGHTSEA’s new Stereo Microscope Fluorescence Adapter (SFA, Figure 1a) now makes it possible to economically adapt both routine laboratory and research level stereo microscopes for fluorescence, streamlining workflow in the research lab for routine tasks such as screening, sorting and fluorescence-guided dissection as well as creating intriguing and scientifically important imaging for undergraduate curricula and research and community outreach.
Figure 1. (a) NIGHTSEA's Stereo Microscope Fluorescence Adapter (SFA) extends fluorescence to routine laboratory or research stereo microscopes for imaging a wide range of animal and plant models including (b) zebrafish (c) fruit fly larvae (d) objects from everyday life such as this burr and (e) the developing root system of the plant, Arabidopsis.

Making Biology courses visual and concrete

For example, a major New England university has created a course integrating genetics, genomics, and evolution to answer the question, “How are observable characteristics of organisms (phenotypes) influenced by genetics?”  This 2nd semester freshman science program is designed to explain the genetic patterns in nature, how genomics can be used to analyze those variations, and how genetics and evolution are intimately related. In the lab portion of the course the SFA is especially useful for examining fluorescently labeled red blood cells and neurons in zebrafish, and the head and muscles of C. elegans.
In a more advanced course, Colgate University goes a step further: visualizing effects of pharmacological agents on zebrafish embryo development.  According to Professor Jason Meyers, “The NIGHTSEA system easily adapted to our dissection scopes (Figure 2).  For quick screens, it worked perfectly well in a bright room.  For more detailed observations, we turned out the room lights.  It worked better than I hoped it would!”
Figure 2.  At Colgate University, undergraduates use the SFA with routine stereo microscopes to observe the impact of pharmacological agents on zebrafish embryo development.

Bringing new light to undergraduate/graduate research

With the SFA, Boston University is able to use the existing teaching lab microscopes in their Genetics Laboratory to enhance research opportunities for seniors and first year Master’s students. In short and long term projects, students formulate and test hypotheses using a variety of genetic techniques that take advantage of fluorescence including mutant selection and screening, chemical genetic screening, complementation, mapping, and recombinant DNA analysis.

Putting a “glow” into community science outreach

Because it is robust and easily set-up (http://bit.ly/sfademovideo), SFA is ideal for the rigors of immersive hands-on science programs as well as community outreach.  For example, the Coastal Marine BioLabs (Ventura, CA, http://www.coastalmarinebiolabs.org) has high school students collect naturally fluorescing marine organisms from the marine  environment (Figure 3), clone the genes encoding both yellow and green fluorescent proteins (YFP and GFP), then deliver the coded genes into specific subsets of embryonic chick spinal cord neurons. According to the program coordinator, “Our goal is not to inspire every student to pursue a scientific career.  Instead, we join a growing alliance of academicians who use the process of scientific inquiry as a tool to foster scientific habits of mind (critical and creative thinking, collaboration, strong communication skills) that will empower our students to become more rational and responsible citizens, environmental stewards, and our next generation of leaders and innovators.”
Figure 3. Coastal Marine BioLabs use the corallimorph Corynactis californica, to involve high school students in authentic, hands-on scientific work.  (a) Brightfield image (b) Fluorescence image made with the NIGHTSEA SFA.
Dr. Jenny Lenkowski of Goucher College loves both teaching and STEM outreach and takes every opportunity to get children and teenagers excited about science. She has a trick up her sleeve:  glowing baby fish (Figure 4).
Figure 4. (a) Dr. Jenny Lenkowski (Goucher College) and a young student observe glowing baby zebrafish under the stereomicroscope. (b) An image of a zebrafish embryo expressing GFP
The glial cells in the zebrafish’s central nervous system readily express green fluorescent protein (GFP).  “The great expression levels in this fish and the mobility of the NIGHTSEA adapter system make it easy to screen fish in a room with the lights on and to take the show on the road to schools and science events like BrainFest.  The children (and the adult family members!) who look through the microscopes were amazed that they could see the nervous systems of live embryos.”
SFA’s versatility and mobility serves a small college like Goucher well.  Systems are shared across multiple labs in independent student projects in introductory biology, upper level courses in developmental biology, and advanced genetics courses.
One of the most comprehensive programs can be found at the University of Montana (Missoula).  Called “spectrUM,” it operates through fixed locations on the U of M campus and a Discovery Center in the heart of downtown Missoula as well as mobile community outreach through MosSE, the Montana spectrUM Science Experience.  The Discovery Center (Figure 5) houses the BrainZone, providing interactive activities for kids of all ages to explore neuroscience.  spectrUM is funded by NIH and staffed by UM researchers and Missoula High School students.
Figure 5.  Using NIGHTSEA’s Stereo Microscope Fluorescence Adapter (SFA) (a) moves the study of fruit fly larvae in the spectrUM BrainZone to the next level.  Selective use of the fluorophore GFP highlights (b) actin filaments which are pervasive throughout the larvae, (c) the peripheral and central nervous system, and (d) salivary glands and cells that will develop into eyes.
“[Our] goal,” cites spectrUM’s director, Holly Truitt, “is to inspire kids to pursue higher education and perhaps careers in science, engineering, math or technology.”

For more information:

Coastal Marine BioLabs – http://www.coastalmarinebiolabs.org


NIGHTSEA develops economical solutions for viewing fluorescence at scales ranging from stereo microscopy to whole organisms. The product range includes a simple system for adding fluorescence to existing stereo microscopes; fluorescence-exciting flashlights and filter glasses; photography accessories, and more. With both off-the-shelf and customized equipment we help thousands of customers around the world in a wide range of applications in research, education, industry, forensic sciences, exploration and more.

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Last updated: May 17, 2017 at 9:06 AM

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