Advanced Cell Analysis Service Center (ACASC)
Birgit Ehmer
To assist the researcher in generating high-resolution, high quality, microscopy-based data for publications and presentation at professional venues. A range of services is available for both experienced and inexperienced users. Experienced users may use the Center's instruments after orientation by a staff member. Inexperienced users may choose to receive training in the use of the instruments, technical support in microscopy and image analysis, consultation in experimental design, or have us perform the microscopy for them as a service.
To book equipment or access services for the ACASC, please login or create an account through Stratocore at https://ppms.us/uc/start/
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Incucyte Zoom (extended live cell microscopy)
Cost:Request Information$50/plate/day
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Digital Macroscopic Imaging (transgenic animals, organs, 2-D gels)
Cost:Request Informationno charge
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Confocal Microscopy (Zeiss LSM 710)
Zeiss LSM710 confocal with 405, 488, 561 and 633 excitation laser lines. Detectors can get configured as needed.
Cost:Request Information$35/hr
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Low Resolution Widefield Fluorescent and Brightfield Microscopy
Cost:Request Information$5/hr
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High resolution fluorescence and brightfield microscopy
Cost:Request Information$15/hr
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Center Staff time
Cost:Request Information$35/hr
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Nikon SIMe super-resolution microscopy.
Super-resolution is available using the 100x objective and the 488nm, 561nm, and 633nm lasers. Wide-field images are available for DAPI-like fluorescence molecule in addition to the super-resolution images.
Cost:Request Information$40/hr
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Flow Cytometry (BD LSR Fortessa)
The Fortessa is equipped with four lasers (405nm, 488nm, 561nm and 640nm) and can detect 2 light scatter parameters (forward and side scatter) and 15 fluorescence parameters. The Fortessa only accepts 5mL polystyrene tubes. Sheath fluid (NERL diluent 2) is provided.
Cost:Request Information$40/hr
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FACS (BD Aria)
Fluorescence activated cell sorter (BD Aria) equipped with four lasers (407nm, 488nm, 561nm and 633nm) and can detect 2 light scatter parameters (forward and side scatter) and 13 fluorescence parameters. Samples can be brought in 5ml or 15ml tubes. Up to 4 population can get sorted. Samples can also get sorted onto tissue culture plates. Sheath fluid (NERL blood bank saline) is provided. $40/hr
Cost:Request Information40/hr without assistance. $75/hr with assistance.
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Confocal Microscopy (Zeiss LSM 710, inverted microscope)
A Zeiss Axio Observer Z1 inverted microscope is connected to a Zeiss LSM710 confocal. The available laser lines are 405, 458, 488, 514, 561 and 633nm. With the availability of a near UV laser this confocal can also visualize DAPI. In addition to the confocal images a DIC image can be acquired. Stage and objective heaters are available to aid live cell imaging.
Request Information
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The most common reason to use a confocal is to obtain optical sections that have much less out-of-focus blur than images from widefield instruments. In addition, one can acquire a 3-D data set for volume determination or 3-D reconstruction.
Multi-tracking provides considerable improvement in the separation of similar dyes over that of a widefield microscope. Therefore, even if you do not need the optical sectioning ability of the confocal microscope, you may want to use it instead of a widefield microscope in order to ensure separation of dye pairs like FITC and rhodamine. Another reason to chose a confocal over a widefield microscope is to have precision in the overlay of images that are taken with different filters. -
Widefield Light Microscopy (multiple stations, upright & inverted platforms)
A Zeiss Axioplan Imaging 2e infinity-corrected upright scope with DIC and epifluorescence. Uses either a Color Zeiss Axiocam (for brightfield imaging) or a B&W Zeiss Axiocam for fluorescence. Filter cubes are available for DAPI, FITC, rhodamine, Texas Red and Cy5-like dyes.
Request Information -
Axiovert 100 TV inverted microscope
This inverted scope is equipped for phase, brightfield, and fluorescence. The filter cubes are suitable for fluoresceine (GFP), rhodamine, and DAPI like dyes. The available objectives are 1.25x, 5x, 10x and a 32x long working distance objective. Additional objectives are available upon request. The filters and optics on this scope are not as advanced as the orcaerzeiss but it is useful for e.g. checking on transfection efficiency. A Zeiss Axiocam MRm is connected connected to the microscope and provides as resolution of up to 1388 x 1040 pixels. The camera is suitable for fluorescence imaging and provides only B&W images.
Request Information -
Stereo Microscope
An Olympus SZX12 stereo microscope serves for low magnification microscopy needs. It is equipped with a Q-imaging color camera and uses Q-Capture software to acquire images.
Request Information -
Incucyte Zoom
IncuCyte ZOOM® consists of a microscope inside a standard cell incubator for environmental stability, and a networked external controller hard drive that collects and processes image data. The instrument has three microscope objectives (4x, 10x, 20x) that can be interchanged by the facility manager upon request.
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It houses up to six microtiter plates which can be run simultaneously. Available imaging modes are phase contrast, red and green fluorescent. Imaging can be for extended periods of time.
Hardware and software tools for a scratch wound migration assay in 96-well plates is available.
Software can be remotely accessed by any networked computer (IP number 10.170.3.126). Ask Birgit for a download location of the program. -
Nikon SIMe
A Nikon SIMe (structured illumination microscopy) superresolution microscope is available on a motorized inverted Nikon Eclipse Ti stand. Available laser lines are 488nm, 561nm and 640nm for acquiring 3D-SIM images. A DAPI channel can be added as a fluorescent widefield channel. The system provides lateral resolution of up to 115nm and axial resolution of up to 269nm.
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The system is equipped with a Hamamatsu Orca Flash 4.0 camera with high sensitivity and low readout noise.
Publications which relied on the equipment:
Chen et al. (2019). Nanoscale monitoring of mitochondria and lysosome interactions for drug screening and discovery. Nano Research 12(5):1009–1015.
Khamo et al. (2019). Optogenetic Delineation of Receptor Tyrosine Kinase Subcircuits in PC12 Cell Differentiation. Cell Chemical Biology 26(3): 400-410.e3
Chen et al. (2018) Super-Resolution Tracking of Mitochondrial Dynamics with An Iridium(III) Luminophore. Small 14 (41): 1802166 -
Flow Cytometer (BD LSRFortessa)
Flow Cytometry is a technology that analyzes multiple physical and/or chemical characteristics of single particles, usually cells, as they flow in a fluid stream through a beam of light. The properties measured include a particle’s relative size, relative granularity or internal complexity, and relative fluorescence intensity.
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The Fortessa is equipped with four lasers (405nm, 488nm, 561nm and 640nm) and detects 2 light scatter parameters (forward and side scatter) and up to 15 fluorescence parameters. The Fortessa only accepts 5mL polystyrene tubes. Sheath fluid (NERL Diluent 2) is provided. -
Fluorescent Activated Cell Sorter (FACS, BD Aria)
Cell sorting is a method to purify cell populations based on the presence or absence of specific characteristics, most often the absence or presence of fluorescent labels. The BD Aria Cell sorter uses in-air droplets to sort. As a collection system we can sort either into two tubes (15ml or 5ml receiving tube with medium) or four tubes (5ml tubes with medium). We also have the option to sort onto tissue culture plates. Due to generation of droplets, the instrument is enclosed in a biosafety cabinet. Sheath is NERL blood bank saline and is provided.
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The available lasers are 407nm, 488nm, 561 nm, and 633nm, and in addition to forward and side scatter up to 13 fluorescent parameters can get acquired.