It is the mission of the Meharry Medical College Morphology Core to provide the expertise and state-of-the-art instrumentation necessary to facilitate and enhance the productivity of Meharry’s investigators and trainees in the areas of fluorescence and bright-field microscopy and immunohistochemistry. This core strives to accomplish this by offering consultation on instrumentation and experimental design, training on instrumentation, and collaboration opportunities. We do our utmost to stay on the cutting edge of technology, skills, and knowledge in the field.
Role: Research assistant
Nikon A1R confocal microscope:
o Live cell imaging with stage top incubator
o Perfect focus
o Dual hybrid scanner for resonant (up to 430 images/s) and galvanic (high resolution) scanning
o 6 laser lines
o Photobeaching capabilities for FRAP, FLIP, FRET, selective photobleaching, and photoconversion or photoactivation
"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."
* 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.
The Olympus BX 50 stereology microscope provides a method to collect data on stained tissue to examine number, length, area, and volume using the Stereologer software. This microscope significantly reduces the time needed to quantify staining of tissue samples, while providing an accurate, unbiased measurement.
Training on instrumentation is provided along with a detailed training user guide. We provide imaging assistance to collect fast and high quality images and data. Technical assistance is available 9am-5pm and other times upon request.
We provide assistance with data analysis and acquiring images.
NIS-Elements performs live cell, 6D experiments quickly and flawlessly. It can organize X, Y, Z, Lambda (wavelength) and Time within one integral platform, for easier-than-ever multi-dimensional imaging. Four types of “dimensions” of acquisition – Time, Multipoint, Z-series, Wavelength (Multichannel)— can be selected.
NIS-Elements innovative analysis capabilities include object counting, automated measurements, region of intent measurements and time measurements including image ratioing of multi-channel images. Advanced visualization methods, like capturing 3D data sets, 2D/3D data set analysis, and large image stitching can all be performed with relative ease with the software. NIS-Elements offers a rich spectrum of mathematical morphology functions (clean, erode, dilate, open, close, smooth), morphologic separation functions, linear morphology functions, fill functions (fill holes, close holes), skeleton functions (medial axis, skeletonize, pruning) and other functions (such as: binary invert, convex hull, contour, skeletonize, homotopic marking, zones of influence, etc.) Multiple single channel images (captured with different optical filters or under different camera settings) can be merged together simply by dragging from one image to another. In addition, the combined images can be stored to a file while maintaining their original bit depths or optionally can be converted into a RGB image.