October 2016: This Month in JoVE

Protocol

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures

Krisztina Janosko¹, Michael R. Holbrook¹, Ricky Adams¹, Jason Barr¹, Laura Bollinger¹, Je T'aime Newton², Corrie Ntiforo², Linda Coe¹, Jiro Wada¹, Daniela Pusl¹, Peter B. Jahrling¹, Jens H. Kuhn¹, Matthew G. Lackemeyer¹

¹Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), ²Environmental Health and Safety, Biological and Chemical Safety Program, University of Texas Medical Branch

Although researchers are generally knowledgeable about procedures and safety precautions required for biosafety level 1 or 2 (BSL-1/2) experiments, they may not be familiar with experimental procedures in BSL-4 suit laboratories. This article provides a detailed visual demonstration of BSL-4 suit laboratory systems check, laboratory entry, movement, and exit procedures.

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 2. General Practices

Steven Mazur, Michael R. Holbrook, Tracey Burdette, Nicole Joselyn, Jason Barr, Daniela Pusl, Laura Bollinger, Linda Coe, Peter B. Jahrling, Matthew G. Lackemeyer, Jiro Wada, Jens H. Kuhn, Krisztina Janosko

Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH)

Performing viral assays in a BSL-4 laboratory is more involved compared to work in a BSL-2 laboratory due to required additional safety precautions. Here, we present an overview of practices and procedures used inside a BSL-4 laboratory illustrating proper Class II biosafety cabinet usage, waste management/disposal, and sample removal.

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 4. Medical Imaging Procedures

Russell Byrum, Lauren Keith, Christopher Bartos, Marisa St. Claire, Matthew G. Lackemeyer, Michael R. Holbrook, Krisztina Janosko, Jason Barr, Daniela Pusl, Laura Bollinger, Jiro Wada, Linda Coe, Lisa E. Hensley, Peter B. Jahrling, Jens H. Kuhn, Margaret R. Lentz

Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH)

Here, we present an overview of the preparation and animal handling procedures required to safely perform medical imaging in an animal biosafety level 4 laboratory. Computed tomography of a mock-infected guinea pig illustrates these procedures that may be used to evaluate the disease caused by a high consequence pathogen.

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 3. Aerobiology

J. Kyle Bohannon, Krisztina Janosko, Michael R. Holbrook, Jason Barr, Daniela Pusl, Laura Bollinger, Linda Coe, Lisa E. Hensley, Peter B. Jahrling, Jiro Wada, Jens H. Kuhn, Matthew G. Lackemeyer

Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH)

As high-consequence pathogens can potentially infect subjects through airborne particles, aerobiology has been increasingly applied in pathogenesis research and medical countermeasure development. We present a detailed visual demonstration of aerobiology procedures during an aerosol challenge in nonhuman primates in an animal biosafety level 4 maximum containment environment.

Investigating the 'Uncatchable Smile' in Leonardo da Vinci's La Bella Principessa: A Comparison with the Mona Lisa and Pollaiuolo's Portrait of a Girl

Alessandro Soranzo, Michelle Newberry

Department of Psychology, Sociology and Politics, Sheffield Hallam University

This paper discusses the methodology used to reveal the 'Uncatchable Smile' illusion in Leonardo da Vinci's La Bella Principessa portrait. A combination of three methods was used (inter-observation, structured interviews, and psychophysical experiments), which led to an investigation that shaped itself without prior beliefs, thus reducing potential researcher bias.

The Use of Induced Somatic Sector Analysis (ISSA) for Studying Genes and Promoters Involved in Wood Formation and Secondary Stem Development

Antanas Spokevicius¹, Lynette Taylor¹, Emma Melder¹, Kim Van Beveren¹, Josquin Tibbits², Nicky Creux^3,4, Gerd Bossinger¹

¹School of Ecosystem and Forest Sciences, Faculty of Science, The University of Melbourne, ²Victorian AgriBiosciences Centre, La Trobe University R&D Park, ³College of Biological Sciences, Department of Plant Biology, University of California, Davis, ⁴Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria

Here we present a protocol that facilitates the medium to high throughput functional characterization of gene and promoter constructs in tree secondary stem tissue within comparatively short time frames. It is efficient, easy to use and widely applicable to a range of tree species.