Vasculata 2018 Workshops

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Instructor: Dr. Steven Segal

Description: This workshop will demonstrate the gluteus maximus muscle preparation for imaging and evaluating the microcirculation of skeletal muscle. The workshop will include an overview of microvascular architecture in skeletal muscle, evaluation of endothelium-dependent vasodilation, adrenergic vasoconstriction, vasomotor tone and microvascular perfusion.

Schedule: 

1:00pm - Brief introduction and orientation (Room M438)
2:00pm - Observe surgical preparation and aspects of intravital imaging (Room 442)
3:30pm - Additional perspective on intravital imaging (M439)
4:30pm - Wrap up and question and answer session (Room m438)

Location: Rooms M438, M442 and M439, Medical Sciences Building, One Hospital Drive

*This workshop can accommodate only 3 attendees

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Instructors: Drs. Michael Davis and Erika Boerman

Description: This workshop will demonstrate the techniques used to microdissect, cannulate, clean and pressurize small arterioles and lymphatic vessels for ex vivo studies. The workshop will include a discussion of methods for manufacturing glass micropipettes and sharpening microsurgical instruments, as well as a demonstration of different systems that can be used to control pressure during an experiment. Opportunities will be available for hands-on experience in dissection and cannulation for each student.

Schedule: 1:00-5:00pm

*This session is limited to only 3 attendees

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Instructors: Dr. Virginia H. Huxley and JianJie Wang

Description: The techniques for making quantitative measures of in vivo microvessel solute permeability and/or hydraulic conductivity will be discussed, demonstrated and opportunities given for hands-on experience by the participants. The workshop will also include a discussion of methods for manufacturing glass microperfusion pipettes and preparation of the solutions of solutes, as well as a demonstration of different systems that can be used to control perfusion during an experiment.

Schedule: 1:00-5:00pm

Location: M455/N408 in Medical Pharmacology & Physiology at MU School of Medicine (next door to Mike Davis' laboratory)

*This session is limited to only 3 attendees

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Instructor: Dr. Bernd Zinselmeyer

Description: This workshop will demonstrate the preparation of popliteal vessels in mice for lymphatic and blood vascular imaging.

Schedule:
10:00-11:20 Demonstration of intravital preparation set up
11:20 – 12:30 lunch break
12:30 – 2:00 Fluorescence imaging lecture including concepts underlying 2-photon microscopy
2:00-2:30 coffee break
2:30 – 5 p.m. live imaging and analysis at the microscope and analysis station

Location: 10:00 - 11:30am - BJCIH 7^th floor, A/B Conference Room, Washington University School of Medicine campus
                12:30 - 5:00pm - BJCIH 7^th floor, Green Room, Washington University School of Medicine campus

Instructor: Dr. Kyunghee Choi

Description: This workshop will introduce the study of embryonic stem cell differentiation into embryoid bodies. Demonstrations will be established to illustrate how the embryoid body system can be used to study vascular sprouting and screen for mediators of this process. It will also demonstrate the use of embryoid bodies for cardiac myocyte differentiation and function. Hands-on experience in seeding embryoid bodies in matrigel and initiating differentiation will be included in the workshop.

Schedule:
1 – 3 p.m. Group 1, EB seeding and differentiation; EB sprouting demo; Cardiac myocyte demo.
3:00 – 3:30 Lecture on ES cells and embryoid bodies.
3:30 – 5:30 Group 2, Group 1, EB seeding and differentiation; EB sprouting demo; Cardiac myocyte demo.

Location: BJCIH 8th floor, Choi Lab space, Washington University School of Medicine

*This workshop can accommodate 10 attendees

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Instructor: Dr. Jessica Wagenseil

Description: This workshop will provide hands on training in biaxial mechanical testing of mouse carotid arteries. The workshop can accommodate 6 students (3 each on two testing machines).

Mouse carotid arteries will be dissected ahead of time and stored in refrigerated saline before use. Participants will mount an artery on a commercial pressure myograph machine (DMT). Participants will inflate the artery to supra-physiologic pressures at different axial stretches and stretch the artery to supra-physiologic axial stretches at different pressures while recording pressure, outer diameter, axial force, and axial length. After testing, participants will cut cross-sections of the artery for measuring the unloaded dimensions and calculating residual stresses. Considerations for determining stress, strain, stiffness, modulus, and energy storage capacity from the testing data will be discussed.

Schedule: 2:00-5:00pm

Location: Urbauer 16 - Danforth campus

Only 1 space left (as of June 21)

Instructor: Dr. Andrew Elvington, Missouri Baptist University

Description: This workshop will review methodology and preparation of vascular casting in a murine model. Attention will be given to cerebral and digestive vasculature but can be altered based on the needs/wants of the attendees. Hands-on participation is encouraged. Uses, analysis, and alternatives will be discussed.

Schedule: 10:00am-2:00pm

10-11am – Discussion of vascular casting: rationale, methodology, considerations
11-12pm – Preparation and administration of specimens/animals
12-1pm – Lunch break
1-2pm – Discussion of imaging modalities and alternatives, continued hands-on participation

Location: BJCIH 11th floor, A/B Conference room Washington University, Medical Campus

Only 1 space left (as of June 20)

Instructor: Dr. Benjamin Humphreys

Description:This workshop will introduce the concepts behind and demonstrate the workflow for single nucleus RNA-seq of mouse aorta using massively parallel single nucleus RNA-seq with droplet technology (DroNc-seq).

Schedule: 1:00-4:00pm

1 – 2 p.m. Lecture reviewing single cell and single nucleus RNA-seq, including dissociation approaches, the major platforms (Drop-seq, InDrops, 10X Chromium) and library creation. Sequencing parameters, costs and basic data analysis and interpretation will be covered as well.
2 – 3 p.m. Demonstration of single nucleus preparation protocol using mouse aorta.
3 – 4 p.m. DroNc-seq run using aorta nuclei.

Location: 9th floor, Wohl Clinic; Washington University School of Medicine

*This workshop can accommodate a maximum of 8 participants.

Instructor: Dr. Brian Wong and Dr. Joel Schilling

Description: This workshop will present an overview of cellular metabolism, discuss methods to assess metabolic parameters in cultured cells, and provide examples of how metabolic assessments are linked to cellular physiology and function. The major metabolic technique demonstrated will be use of the Seahorse and interpretation of data generated.

Schedule:

Time	Topic	Type
10:00-10:30AM	Metabolism   overview (BW)	Lecture
10:30-11:00AM	Seahorse   theory (JS)	Lecture
11:00AM-12:30PM	Seahorse   practical	Demo
12:30-2:00PM	Lunch   break
2:00-3:00PM	Seahorse   analysis	Demo
3:00-3:20PM	Endothelial   metabolism in health and disease (BW)	Lecture
3:20-3:40PM	Immunometabolism   in health and disease (JS)	Lecture

Location: BJCIH 8th floor, Green Room, Washington University School of Medicine campus

Leaders: Monica Shokeen, Ph.D., Optical Imaging Lab, Radiology; and Dana R. Abendschein, Ph.D., Internal Medicine/Cardiology, Washington University School of Medicine in St. Louis

Schedule:
10-11: Measuring Angiogenesis with PET/CT and Complex Atherosclerosis/Vascular Injury with PET/MRI
and Optical Imaging – Dana
Animal models of occlusion and hind limb ischemia will be discussed/demonstrated
11-12: Assessing Vascularity and Bone Marrow Composition in Multiple Myeloma with Optical, Nuclear and Magnetic Resonance Imaging – Monica
Pros and cons of various animal models for studying hematological malignancies will be discussed
12-1: Lunch Break
1-2:30: Field Trip to MicroPET/MR and Optical facilities in MIR and Biomedical Couch Building (Optical Radiology Laboratory)
2:45-5:00: We will include hands on exercises in image analysis. View microPET image analysis in Optical Imaging Lab, Couch Building and/or have additional topical discussions with WU investigators. Possibilities include alpha v beta 3 targeting for imaging (Dr. Weilbaecher), brain imaging (Dr. Corbo), Culver group for brain vasculature imaging (oxy/deoxy) and/or other applications as participants have interest (Dana or Monica to facilitate connections).

Location: ORL Conference Rooms, Couch Biomedical Research Building, 4525 McKinley Ave, 2nd Floor

*This workshop can accommodate 15 attendees

Instructors: Drs. Saulius Sumanas and Dr. Andrew Koenig

Description: This workshop will introduce basic techniques used to study vascular development and function in zebrafish embryos. Brief demonstration will be followed by hands-on exercises.
The workshop will cover the following: overview of zebrafish vascular development and basic embryo handling techniques; phenotypic analysis of vascular defects in different mutants using fluorescent reporter lines; treatments with chemical inhibitors and analysis of vascular defects; embryo microinjection and microangiography; heme staining and analysis for hemorrhages. 

Time: Wednesday, July 25, 2018; 10-3 pm (lunch break 12-1)

Location: Zebrafish Consortium Facility, Washington University School of Medicine, McDonnell building, basement

*This workshop can accommodate 10 students.

 

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Instructor: Jesse Williams, PhD, Dept. of Pathology and Immunology, Washington University School of Medicine

Schedule:
10-11:30 am: Demonstration and hands-on experience in isolation of aorta; including ascending, descending, abdominal, and carotid branches from atherosclerotic mice.
11:30-1:00 pm: Digestion protocol to separate aortic adventitia from the intima/media, followed by single cell suspension preparation
1:00-2:30 pm: Lunch Break
2:30-3:30 pm: Flow cytometric leukocyte staining and introduction to flow cytometry
3:30-5:30 pm: Field trip: Washington University Flow Cytometry Core, group training on flow cytometer (BD LSRFORTESSA X-20) setup, compensation (FACS DIVA), and gating strategies (FLOWJO).

Location: BJCIH 8th floor, Randolph Lab space

*This workshop can accommodate 5 attendees

Instructor: Dr. Gwen Randolph

Description: The writing workshop will be set up from 10:00 a.m. to 5 p.m. on a come-and-go basis, so that conference attendees can join during times they may have available around other workshops attended. Dr. Gwen Randolph will make available lessons and expertise on scientific writing as the main goal of the workshop, including manuscript and research proposal writing. Material from the lectures in the previous two days will serve as a source material to learn to write professional and persuasive scientific documents. Students may also bring topics from their own research to work on. Participants will be able to come and go as they please. If you are in another workshop that is perhaps just a half day, you can participate in this workshop the other half (except for those going to the University of Missouri).

Schedule: 10:00am - 5:00pm

Location: BJCIH 8th floor, A/B Conference Room