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Each piece of equipment available to CEG members through the ITS Core has an online scheduler. To access, first click on the type of equipment (links in red below), then read and click on the Sign-up\reservation form.
Samples should be prepared with 96-well sample plates (PDF) (U-bottom preferred) or 1.5ml sample tubes (PDF). The cell suspension should be maintained at ~200 cell/ml and ~200 ul/well for the 96-well plate format.
A project drive, called “Document-Flow users”, has been set up for temporary storage. If the computer space is limited, the stored data will be removed after 6 months from initial acquisition.
Data can be analyzed with the software GuavaSoft inCyte or Guava Suite provided by the manufacturer. Data can also be analyzed with FlowJo (group license is available from CCHMC flow cytometry core), FCS Express, and ModFit LT, which are also available for data analysis free of charge to CCHMC flow core users in R5503. See "Software" for more information.
Users are encouraged to understand the basic knowledge of flow cytometry in experimental design, antibody staining, compensation, and data analyses together with unique setting for this easyCyte unit to ensure success. These resources include the instrument website of Guava easyCyte 12-channel high throughput (12HT) flow cytometer, flow cytometry resources, on campus lectures provided by Ohio River Valley Cytometry Association (ORVCA), and related seminars from CEG.
Contact: Shouxiong Huang (Kettering Rm 248, Rm 251), Shouxiong.huang@uc.edu; Manju Sharma (Kettering Rm 248, Rm 276) SHARMAMU@ucmail.uc.edu
The instrument is located in Rm 253 Kettering
Training mainly involves basic instrument and software manipulation to allow new users to start sample measurement independently. At least 1 hour of instrument demonstration and 1 hour guided hands-on sample running are required prior to independent usage. Users can request more training hours if they are unable to fully perform the assays independently.
Trained users can reserve and access the instrument from 8:00 am to 5:00 pm, Monday through Friday. Usage may be available after working hours at a case-by-case scheduling basis Users are responsible for the correct manipulation of the instrument to achieve results and avoid damages.
A staff is available to run pre-prepared samples. This service is currently offered at a restricted time. Currently, the service is open every week for two-three hours slots on Monday and Wednesday afternoons.
Users are encouraged to make appointments with Dr. Huang to discuss experimental design, pitfalls, and data analyses.
Contact: Shouxiong Huang, PhD
Research supported by the Center for Environmental Genetics should acknowledge NIEHS Award P30 ES006096.
Sign up using the registration form
ImmunoSpot® Analyzers are designed for scanning and evaluating single cell-based spots, which reflect the protein expression, function, and growth of cells. Technically, the ImmunoSpot® S6 Micro’s zoom lens permits seamless switching from 6- to 384-well plates and counting objects <10 µm in size – including individual cells. With its maintenance-free 150-diode overhead lighting system and proprietary planar backlight system, the ImmunoSpot® S6 Micro provides the ultimate in image quality, especially for double-color assays (no fluorescence signal detected by this model). It is a low maintenance and high performance instrument for measuring functional responses and clustering growth derived from single cells at a high-throughput plate-based setting.
This ImmunoSpot® S6 Micro Analyzer can be applied to the following CEG-related or supported studies involving in environmentally induced diseases and conditions. (i) Characterization of cytokine, surface markers, or soluble molecules from rare cancer stem cells with high sensitivity and accuracy to capture individual cells. (ii) Functional measurement of the cytokine production from rare immune cell subsets, e.g. regulatory T cells, anti-inflammatory immune cells, etc, from human samples in response to toxicant-treatment or environmental exposure. (iii) Many reagents kits have been commercially developed for testing cytokine production that can be conveniently applied in toxicology studies. (iv) The growth of microbes from various pathogenic sources or from complex microbiota pools can be recorded, quantified, and differentiated using the high-resolution camera and analytical software, which can be applied for studying the environmental toxicity on microbiota. Notably, the ImmunoSpot® Analyzer is highly advantageous in functional characterization of rare cells from limited human samples and in the survey of rare subsets from exposure-related epidemiology studies using high-throughput microplate analyses.
Contact: Shouxiong Huang (Kettering Rm 248, Rm 251), Shouxiong.huang@uc.edu
Location: The instrument is located inRoom 253 Kettering Laboratory Building
Training mainly involves basic instrument and software manipulation to allow new users to start sample measurement independently. Users can access online resources from manufacturers or vendors for available reagents, general protocols, and overall manipulations. At least one-hour instrument demonstration and hands-on sample running are required prior to independent usage. Users can request more training hours if they are unable to fully perform the assays independently.
Trained users can reserve and access the instrument from 9:00 AM to 5:00 PM, Monday through Friday. Usage may be available after working hours at a case-by-case scheduling base. Users are responsible for the correct manipulation of the instrument to achieve results and avoid damages.
Consultation Users are encouraged to make appointments with Dr. Huang to discuss experimental design, pitfalls, and data analyses. Contact: Shouxiong Huang, PhD
Sign up using the Registration Form
The Confocal Imaging service is equipped with “Carl Zeiss Confocal Laser Scanning Microscopy” LSM700.
Confocal Laser Scanning Microscopy combines high-resolution and high-contrast optical imaging with depth selectivity allowing visual sectioning of tiny structures and constructing three-dimensional (3D) structures from the obtained images. It is broadly used to resolve the detailed structure of specific objects within the cell; it also enables the creation of sharp images of the exact plan of focus without any disturbing fluorescent light from the background or other region of specimen. Stacking several images from different optical plans allows visualization and analyses of 3D structures. However, due to limited depth of sample penetration, the thickness of specimen is an important consideration for experimental success. Use of confocal microscopy in the biomedical sciences includes the imaging of multiple labeled specimens and the measurement of physiological events in living cells. Visualization of sufficient variants within both cell and tissue context will allow spatial and dynamic analyses of cell biology and may lead to advancement in understanding of biological processes.
Brochure
Contact: Chia-I Ko, Ph.D. (koci@ucmail.uc.edu)
Location: Kettering laboratory complex, Room 443 (4th floor, Kettering Addition).
Instruction: Microscope use is free of charge only for CEG members. The sign-up/reservation form is mandatory for each use (link under development; in the interim, please contact Dr. Ko). Individuals who have received the microscope training are allowed and will be approved to schedule a reservation. New users should contact Dr. Ko to schedule an initial training before planning for experiments. Other services such as sample preparation instructions and troubleshooting for image-taking can be provided upon request; however, we do not guarantee for expected results. In case system failure is detected due to carelessness and mishandling, users will be charged for repair costs.
Odyssey imaging Systems: The Odyssey CLx is the next generation multifunctional imaging platform that can provide a wide range of applications. The Odyssey CLx platform uses infrared laser excitation that out-performs LED and visible white light systems for Western blots. This increased sensitivity offers a clear image of your data that is unmatched by other digital imaging systems. Two separate lasers and detectors simultaneously detect both fluorescent signals. The optical system employs diode lasers and solid-state detectors with long lifetimes and very low maintenance requirements. Infrared laser excitation outperforms systems that use white light, LED light sources, and filter wheels by delivering higher intensity excitation light to the fluorophore. A variety of fluorescent dyes and stains are compatible with the 700 nm and 800 nm excitation wavelengths of the two diode lasers in the Odyssey CLx. Spectral overlap is minimized by a 100 nm separation of the two detection channels, and optical filtering ensures that each detector measures fluorescence from only one of the infrared dyes.
Major advantages
Training: Training will mainly involve basic instrument and software manipulation to allow new users to start taking western-blot images independently. At least 10 min instrument demonstration and first time guided hands-on software running are required prior to independent usage. For Odyssey-CLx knowledge, experimental design, data interpretation, and result presentation, users should refer to tutorials available on request. Also refer to https://www.licor.com/bio/webinars/ , https://www.licor.com/bio/odyssey-clx/resources before starting experiment.
Scheduling: After being trained and able to perform assays independently, users can contact our staff for scheduling. Usage slots of more than one hour must be justified. Users are also required to sign on the usage page and provide project information for CEG.
Sample format: Western blot membranes and reagents should be provided by users. First time users of this NIR western blotting technique can request use of secondary antibodies and blocking solutions from Dr. Tarapore.
Fees: Available to CEG members without charge
Data Storage: A project drive, called “CEG_Users”, has been set up for temporary storage. If the computer space is limited, the stored data will be removed after 6-month from initial acquisition.
Data Analysis: Data can be analyzed with the software Image Studio provided by the manufacturer. To contact Dr. Ying Xia for details click on this email link.
Description: Agilent Seahorse XF HS Mini Analyzers measure mitochondrial respiration and glycolysis as well as ATP production rate of live cells in an eight-well miniplate format. The quick and easy setup makes the Seahorse XF HS Mini Analyzer ideal for performing routine energy metabolism measurements of ex-vivo and other quantity-limited samples.
Detailed information can be found at: https://www.agilent.com/en/promotions/seahorse-xf-hs-mini-analyzer
Sign-up/reservation is required. New users must complete Agilent’s webinar-based Seahorse training prior to use. Please follow this link to register. For additional questions about training, please contact Kathyrn or Jestine, Applications Engineers- Cell Analysis Division, Agilent Technologies, Inc. Email: kathryn.tormos@agilent.com Email: jestine.ho@agilent.com
Location: Kettering Laboratory Complex, Room 225D
Description: The Optima MAX-TL microprocessor-controlled tabletop ultracentrifuge can generate high centrifugal forces up to 120,000 rpm / 627,000 g, suitable for small volume samples. Manual and programmed operations are available from the integrated touchscreen interface. An ultracentrifuge is capable of separating high-molecular-weight compounds, viruses and subcellular particles for study. Two Beckman rotors are available.
• TLA-120.2, fixed angle rotor for 10 x 1.0 mL open top tubes, max speed 120,000 rpm (627,000 g). • TLS-55, swinging bucket rotor for 4 x 1.0 or 2.0 mL open top tubes, max speed 55,000 rpm (259,000 g). Detailed information can be found at • https://www.beckman.com/centrifuges/ultracentrifuges/optima-max-tl# • https://www.beckman.com/centrifuges/rotors/fixed-angle/357656 • https://www.beckman.com/centrifuges/rotors/swinging-bucket/346936 Sign-up/reservation is required. New users should schedule an initial training. The training includes a walk through of centrifuge parameter set up, rotor handling and before and post usage care, cleaning, and sterilization. For trained users, the instrument is open for use at any time of the day, but usage outside working hours needs to be specially arranged with the manager for access. Reservation should be made between 9:00 a.m to 5:00 p.m Monday through Friday, at least 4 hours prior to the planed usage and rotor/tube pick up. Contact information: Bo Xiao (xiaobo@mail.uc.edu), Kettering Lab 410
Location: Optima Max TL ultracentrifuge is located in the Kettering Laboratory Building, Room 411
CSign up using the Registration Form
Description. The RC6 Plus is a high-speed (to 55200 g / 22,000 rpm) centrifuge used to separate substances of different densities at controlled temperatures. Running conditions are selected with the touch keyboard. Actual and set run conditions are displayed. Two Thermo Scientific Superspeed rotors (i.e., F21-8x50, HB-6) are available.
• F21-8x50, fixed angel rotor for 8 x 50 mL tubes, max speed 52,356 g (21,000 rpm) • HB-6, swinging bucket rotor for 6 x 50 mL tubes, max speed 27,617 g (13,000 rpm) Detailed descriptions can be found at
Location: Centrifuge is located at Kettering laboratory complex, Room 419
Description: The Environmental Simulating Chamber for Aerosol & Particle Exposure (ESCAPE) is a modular inhalation chamber system that can cater to users’ needs to study various types of aerosols/particles with both in-vitro (air-liquid interface) and in-vivo (mice/rats) assays. ESCAPE has five sections:
Contact: Jun Wang, wang6ju@ucmail.uc.edu, 331 Kettering; Jagjit Yadav, yadavjs@ucmail.uc.edu, 137 Kettering
Location: 172 Kettering Lab Complex
Description: The EVOS® FL Auto Imaging System is designed to eliminate the complexities of high-end microscopy while maintaining the highest levels of performance. This fully automated multichannel fluorescence imaging system was developed for advanced automated imaging applications such as time-lapse imaging, multi-well plate scanning, image stitching and tiling, and cell counting. You can quickly image a sample to check its condition or experimental progress, or use the EVOS ® FL Auto Imaging System equipped with the optional onstage incubator to acquire high-resolution time-lapse movies of experiments spanning many hours.
Features and applications of the EVOS® FL Auto Imaging System include
Contact: Chia-I Ko, PhD. Email: koci@ucmail.uc.edu
Location: 443 Kettering Laboratory
EVOS is available for sign-up in 2-hour time slots, Monday through Friday, 10:00 AM - 4:00 PM. Reservations must be made 2 days in advance. Please contact Dr. Ko for any early morning or late evening emergency usage requests.
Description: RNAScope is a commercially available RNA in situ hybridization (ISH) that allows visualization of single RNA molecules in individual cells in a variety of sample types including formalin fixed paraffin-embedded tissue sections.
The ACD HybEZ II Hybridization System is a hybridization incubator specially designed for RNAScope. The incubator holds up to 20 slides in a rack and provides the necessary temperature control and humidity required for optimal performance of RNAscope assay. The system comprises: HybEZ oven, EZ-Batch Wash Tray and EZ-Batch Slide Holder.
• Temperature range: (ambient + 10℃) to 75℃ • Setting range: 20℃ to 75℃
Click here for details about the ACD HybEZ II Hybridization System 110v Click here for details about the Hyybez II System for Manual Assays
Sign-up/reservation is required. New users should schedule an initial training. The training includes a walkthrough of oven parameter set up, tray handling and before and post usage care and cleaning.
For trained users, the instrument is open for use at any time of the day. Reservation should be made between 9:00 a.m. to 5:00 p.m. Monday through Friday, at least 4 hours prior to the planed usage. If room access outside of working hours is required, contact the manager for special arrangements. Contact information: Ying Xia, PhD; xiay@ucmail.uc.edu, 410 Kettering Laboratory Complex
Location: The oven is location in Kettering room 411.
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Published work supported by the Center for Environmental Genetics or featuring data derived by CEG Cores must comply with the NIH Public Access Policy at the time an article is accepted for publication. Published work also must acknowledge NIEHS award P30ES006096. Answers to questions about these duties may be found at https://publicaccess.nih.gov/faq.htm
Access papers into Pub Med Central
Authors with work supported in full or in part by a CEG award or CEG Core should verify that the P30 ES006096 award is associated with their publication in MyNCBI\MyBibliography after the publication is marked as Compliant or at least In Process for Pub Med Central (PMCID): https://account.ncbi.nlm.nih.gov/
Users can find a helpful View 3-minute tutorial