Resources and Contacts
Resources are available to do immunostaining of cells and fixed tissue*. The researcher is responsible for providing the primary and secondary antibody. The Core also houses state-of-the-art microscopes capable of handling the diverse imaging needs of its users included but not limited to wide-field, confocal, bright-field, fluorescence, calcium ratio imaging, and stereology.
NOTE: *If tissue sectioning is needed, frozen and paraffin sections can be prepared in the Tissue Acquisition Core
Shandon Cadenza automated multi-slide immunostainer
- This is a fully programmable automated immunostainer, designed to perform all stages of immunostaining (ICC) from the application of blocking agents up to and including counterstaining.
- It can accommodate up to 9 different primary antisera.
- It is capable of processing 20 slides at one time.
Instrumentation:
Nikon TE2000-U C1 confocal laser scanning microscope
- This confocal has 488 nm, 543 nm, and 633 nm laser lines to excite fluorophores with compatible excitation characteristics. Fluorescence emission is collected with 505-550 nm, 560-615 nm, and 650 nm filter sets, respectively, in independent channels. The microscope is also equipped with a fourth detector to collect light for differential interference contrast (DIC) imaging.
- There are 10x, 20x, 40x, 60x, and 100x objective lenses available for use on this microscope. They are all capable of imaging using DIC, making it possible to do brightfield confocal imaging.
- Motorized z-plane movement allows for optical sectioning through a sample. The EZ C1 confocal software can be used subsequently to reconstruct serial z-plane images into a 3D rendering.
- Images obtained with this microscope
Confocal Image
Nikon TE2000-E is a fully automated wide-field fluorescence microscope. The TE2000-E wide-field microscope is equipped for diverse imaging:
- Motorized z-focusing motor enables collection of in focus images in different image planes. This utility is most often used in acquiring images for deconvolution and Stereolgy.
- It has a xenon lamp for fluorescence excitation provides an excellent light source for calcium imaging with Fura-2 and UV-Vis fluorophores.
- A very sensitive CoolSNAP HQ2 camera is ideal for fluorescence imaging, as it produces very little noise and allows for faster integration times.
- A 5 mega pixel color camera is available for non-fluorescent samples and is ideal for histological stains.
- There are 4x and 10x phase lenses available as well as high NA 20x, 40x, 60x, and 100x DIC lenses.
- A computer controlled excitation filter wheel allows for fast, automated collection of images.
- Filter sets are available for UV, blue, green, and red excitation and emitted light is collected with the corresponding band pass or long pass emission filters.
- Filter sets are available for calcium ratio imaging, specifically fura-2 AM.
- A motorized stage in conjuction with Nikon NIS Elements or Stereologer software allows for the collection of images in multiple, specified locations.
- Nikon NIS Elements Advanced Research imaging software is cable of 6 dimensions (6D) of acquisition. Up to six combinations of wavelength, time, x-, y-, and/or z-positions are possible to control with this software.
- Images obtained with this microscope
Wide-Field image
Calcium flux images
Scientific Director: Sukhbir S. Mokha, Ph.D.
- Office: B47 Biomedical Center
- Phone: 615.327.6933
- Dr. Mohka ensures that the Core meets the needs of the SNRP program, the principal resource funding the Core, and is working toward anticipated future needs of Meharry investigators. He and Dr. Goodwin work closely together to make sure that the needs of the users are fulfilled.
Managing Director: J. Shawn Goodwin, Ph.D.
- Office: 3109 West Basic Sciences
- Phone: 615.327.6686
- Dr. Goodwin oversees the operation of the Core facility. His expertise is in quantitative, live-cell imaging; however, he is knowledgeable in all aspects of the Core. He is available for consultation on projects and planning of experiments, and is willing to collaborate with investigators, if appropriate. He also conducts training on Core equipment for independent use.
Consultant: Ian H. Fentie, Ph.D.
- Office: 3109 West Basic Sciences
- Phone: 615.327.6686
- Dr. Fentie is a resource for immunostaining of tissue and cells. He also is available to train researchers for independence on Core microscopes and collaborate on research projects.
- Morphology Monday Clinics: Bi-weekly meetings to discuss topics of interest relating to immunohisto- and immunocytochemistry and microscopy. These meetings are held from 12-1 pm in the Office for Research Conference Room (2011 WBS). Lunch is provided to attendees with RSVP three days prior to the meeting.
- Immunolabeling techniques.
- What is fluorescence?
- How to collect quantitative data and analyze it.
- Proper techniques to collect high quality, multi-labeled fluorescence data.
- How to collect live-cell images with the confocal microscope.
- How to conduct calcium imaging experiments.
- Quantitative live-cell techniques.
- How to do Stereology properly.
- Upcoming regional/national meetings of interest to users:
- Microscopy Mini-Course May 14-18 at VUMC
- Lab assignments
- Topics covered include
- Transmitted Light & Fluorescence Microscopy
- Quantitative Digital Imaging
- Confocal Microscopy & Related Techniques
- Live Specimen Imaging & Advanced Techniques
- Electron Microscopy & Correlative Imaging
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