Instrumentation Training
Several avenues for individuals to be trained on instrumentation and techniques commonly used in the IRF exist. The most direct is to schedule time with the appropriate staff member for individual training on a piece of equipment. All staff members have a number of responsibilities, so please do not complete an experiment and request immediate training or help. Please schedule time for assistance at least one week in advance.
Course Credit
Two avenues exist for receiving graduate credit while learning to use equipment in the IRF. The first is through the Independent Studies mechanism with one of the faculty members. This involves extensive training on an instrument or technique including sample preparation and the completion of a research project.
The second avenue for receiving graduate credit is through the MCBA 740 and 741 courses offered through the Department of Cell Biology and Anatomy. Participants in these courses spend two to three weeks consisting of lecture and laboratory hours on several pieces of equipment in the IRF. The goal of the courses is to provide a basic knowledge of several pieces of equipment and the types of data that can be obtained from the equipment. Laboratories are hands-on and students are expected to complete a series of basic sample preparations and to collect and analyze data from a series of assigned exercises. Flexibility in the instrumentation covered each semester exists and the courses can be modified based on the needs of the students enrolled. A brief written report is required following each unit of the course.
MCBA 740 (Fall Semester) – Structural Imaging Methods in Biomedical Research
Topics include: histological techniques, light, confocal and electron microscopy, digital imaging and ethics
MCBA 740 Syllabus
Lecture
Intro: Price
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Course Requirements
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Rules for use of IRF
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Specimen fixation, processing, sectioning, H&E and Trichrome stains
Lab
Schedule time in groups of 2 with Benny Davidson for training
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Fixation and Processing to get paraffin blocks
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Paraffin and Cryo sectioning
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H&E and Trichrome stains on tissue of choice
Lecture – Price
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Antigen: Antibody Interactions, Immunocytochemistry and Histochemistry
Lab
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Continue with above lab material – Benny
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Use Antigen Retrieval techniques on paraffin slides and stain slides with provided immunohistochemistry kit – Sharon Cooper
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Training on light microscopes and image acquisition – Dr. Fuseler
No lecture – Labor Day
Lab
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Continue with above lab exercises -Davidson
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Use of Automated staining and Image Acquisition systems – Sharon Cooper
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Photoshop – Jeff Davis
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Image Pro Plus – Jeff (Alternatively if you prefer to learn Metamorph you can work with Dr. Fuseler on this program)
Lecture – Price
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Antigen: Antibody interactions for histochemistry/cytochemistry
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Digital Imaging
Lab
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Finish with above materials. You should be done with all sectioning, staining and image acquisition exercises by the end of week 4.
Lecture - Price
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Digital Imaging Continued
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Confocal
Lab
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Specimen processing for confocal including cells, Vibratome sections and triple label staining – Anna Harper
Lecture – Price
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Confocal Continued
Lab – Price and Anna Harper
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Confocal training - Price
Lecture – Price
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Confocal continued
Lab
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Confocal continued
Lecture
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Confocal continued
Lab
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Confocal continued: All confocal work should be finished by the end of Week 8
Lecture – Fuseler
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Live cell imaging
Lab – Fuseler
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Time lapse imaging
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Imaging calcium/pH fluxes
Lecture – Price
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Scanning and Transmission Electron Microscopy
Lab
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Finish live cell imaging exercises - Fuseler
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Start processing specimens for SEM and TEM - Jeff
Lecture – Price
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Scanning and Transmission Electron Microscopy
Lab – Price, Fuseler and Jeff Davis
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Finish processing specimens for SEM and TEM - Jeff
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Training on SEM – Jeff
Lecture – Cell Ultrastructure
Lab
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Image SEM samples – Jeff
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Ultramicrotomy demonstration – Price
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Staining TEM samples – Jeff
Lecture – none
Lab
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Training on TEM – Price
Lecture – none
Lab
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SEM and TEM exercises
Finish all labs
MCBA 741 (Spring Semester) – Proteomic and Genomic Imaging Methods in Biomedical Research
Topics include: BioPlex/Luminex technology, laser capture microdissection, real time
PCR, flow cytometry and cell sorting, mass spectrometry and DNA microarrays
MCBA 741 Syllabus
Lecture
Price: Intro
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Introduction to the IRF
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Rules for use of IRF
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Course Requirements
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Potts: RNA techniques: In situ hybridization, Agilent 2100 Bioanalyzer, Real time PCR
Lab
Potts – Schedule time Dr. Potts for training on equipment
No lecture – MLK Day
Lab
Continue with above lab material
Lecture – Potts
RNA techniques: In situ hybridization, Agilent 2100 Bioanalyzer, Real time PCR
Lab
Continue with above lab exercises
Lecture – Potts
RNA techniques: In situ hybridization, Agilent 2100 Bioanalyzer, Real time PCR; Begin protein analysis lectures
Lab
Continue with above lab exercises and schedule time with Dr. Potts for protein analysis exercises
Lecture – Potts
Protein Techniques: Western blots, Phosphorimager, Bioanalyzer
Lab
Finish all above exercises for in situ hybridization, Agilent 2100 Bioanalyzer and Real Time PCR
Schedule time with Dr. Potts for protein analysis exercises
Lecture - Valarmathi
Stem cell techniques
Lab
Continue with protein analysis and schedule time with Dr. V for stem cell exercises
Lecture – Singh
FACS and Cell Sorting Techniques: FC 500, FACS ARIA II, AutoMACS Pro
Lab
Continue with Stem Cell Exercises and schedule time with Dr. Udai Singh to begin cell sorting exercises
Lecture – Singh
Cell sorting techniques:
Lab
Continue with Stem Cell and cell sorting exercises
Lecture – Singh
Cell sorting techniques
Lab
Continue with and cell sorting exercises
Lecture – Singh
Cell sorting techniques
Lab
Continue with Stem Cell and cell sorting exercises
Lecture - Singh
BioPlex and cell sorting
Lab
Finish FACS and sorting exercises
Schedule time with Dr. Singh on BioPlex
Lecture – Singh
BioPlex and Cell Sorting
Lab – Singh
Continue with experiments on FACS and BioPlex equipment
Lecture – Creek
Microarray technology
Lab – Singh and Creek
Continue with experiments on FACS and BioPlex equipment
Schedule time with Dr. Creek for microarray exercises (equipment and lab in Pharmacy on main campus)
Lecture – Creek
Microarrays if necessary
Lab – Creek
Continue with experiments on microarray technology
No Lecture
Lab
Finish microarray exercises