Research Core Facility
The Research Core Facility at LSU Health Shreveport provides investigators, post-doctoral fellows, and graduate and undergraduate students with access to a wide variety of research services, educational and training opportunities and state-of-the-art technologies.
Located on the 6th floor of the Biomedical Research Institute, each technology is staffed by a trained research specialist. These technologies include cellular metabolism, flow cytometry, laser capture microdissection, mass spectrometry, microarray, fluorescence, confocal, and super resolution confocal microscopy, next generation sequencing, and real-time PCR. The Research Core Facility provides assistance with all aspects of these technologies, including experimental design, protocol optimization, instrument operation and data analysis.
We welcome investigators on the LSU Health Shreveport campus, as well as those associated with other campus, to utilize any of these technologies on a nominal fee-for-service basis. We encourage you to contact us for more information.
Using the Research Core Facility
To use any technology in the RCF, please consult with staff to discuss sample preparation, scheduling, fees and training.
Training
All users must be trained by Research Core Facility staff. Training by other users is not sufficient. Anyone who operates or attempts to operate RCF equipment without proper training will be subject to disciplinary action such as loss of future access to all RCF technologies.
Depending on the specific technology, training may involve anything from one or more one-on-one sessions with a staff member to formal classes developed by RCF staff. Please contact RCF staff to schedule training on any end-user operated instrument in the Core. Please note that training will NOT be done on a drop-in basis and MUST be scheduled ahead of time.
Data Management
Responsibility for the storage of data generated from experiments or samples analyzed in the RCF belongs to the investigator. Each investigator (or his or her staff) is provided with printed and/or electronic copies of all analyses conducted by RCF staff. Each investigator must ensure that data is properly stored and protected in his or her laboratory.
The RCF will make every effort to retain duplicate copies of data and will store them on electronic media for a period of three years. In the even an investigator cannot locate the original data, he or she should contact RCF staff to obtain a duplicate copy. In cases where data are generated by the investigator on end-user operated instruments, the individual who runs the experiment must immediately make an electronic copy of data and keep that in the investigator's lab. Although the RCF assumes no responsibility for data files left on end-user RCF computers, stall will make every effort to periodically back them up.
Equipment List
Research Core Facility Equipment List:
updated November 2020
Cellular Metabolism
Agilent Seahorse XFe24 Analyzer
Flow Cytometry
ACEA NovoCyte Quanteon Flow Cytometer (2)
BD Biosciences FACSAriaIII Flow Cytometer/Cell Sorter
BD Biosciences LSRII Flow Cytometer
Laser Capture Microdissection
Arcturus XT LCM System
Mass Spectrometry
Thermo Scientific Orbitrap Exploris 480 Mass Spectrometer
Microarray
Affymetrix GeneChip System
Microscopy
Leica TCS SP5 Confocal Microscope
Nikon A1R Confocal & N-SIM Super Resolution Microscope
Nikon Eclipse TE300 Widefield Microscope
Olympus iXplore CSU W1 Spinning Disk Confocal Microscope
Zeiss AxioObserver Z1 Widefield/Apotome Microscope
Zeiss LSM 510 NLO Confocal/Multiphoton Microscope
Next Generation Sequencing
Illumina MiSeq Next Generation Sequencer
Illumina NextSeq 550 Next Generation Sequencer
Real-Time Quantitative PCR
Bio-Rad CFX96 Fast Touch Real-Time PCR Instrument (x3)
Bio-Rad CFX384 Touch Real-Time PCR Instrument
Equipment Descriptions
- Cellular Metabolism
- Flow Cytometry
- Laser Capture Microdissection
- Mass Spectrometry
- Microarray
- microscopy
- next generation sequencing
- Real-Time PCR
Cellular Metabolism
The Agilent Seahorse XFe24 Analyzer measures the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of live cells in a 24-well plate format. OCR and ECAR rates are key indicators of mitochondrial respiration and glycolysis as well as ATP production rate, and together these measurements provide a systems-level view of cellular metabolic function in cultured cells and ex-vivo samples.
Flow Cytometry
ACEA NovoCyte Quanteon
The ACEA NovoCyte Quanteon analyzer is a 4-laser flow cytometer capable of detecting up to 27 parameters (forward scatter, side scatter, and 25 fluorescence detectors). In addition to a 405nm, 488n, and 637nm laser, it includes a 561nm laser, which provides the ability to excite yellow and green fluorophores of conjugated antibodies.
BD Biosciences FACSAriaIII
The FACSAriaIII cell sorter is capable of 17-parameter (Forward Scatter, Side Scatter, and 15 fluorescence detectors) analysis and cell sorting. It has four solid-state lasers for excitation at wavelengths of 405 nm, 488 nm, 561 nm and 633 nm. It is a digital high-speed sorter, capable of sorting up to 70,000 events per second. It is capable of standard “bulk” sorting of up to four user-defined cell populations simultaneously, or can be used for direct deposition of a counted number of cells directly into tissue culture plates for cloning, frequency response assays, or other single cell analyses. This instrument uses BD Biosciences FacsDiva software for acquisition and analysis.
BD Biosciences LSRII-SORP
This BD LSRII is capable of detecting up to 17 parameters (Forward Scatter, Side Scatter, and fifteen fluorescence detectors). It has four solid-state lasers for excitation at wavelengths of 405 nm, 488 nm, 561 nm and 633 nm. This instrument uses BD Biosciences FacsDiva software for acquisition and analysis.
Laser Capture Microdissection
The Arcturus XT Laser Capture Microdissection System performs laser capture microdissection from heterogeneous tissue samples simply, quickly, and precisely. In minutes the investigator can locate a single cell or large groups of cells and, using a simple aim-and-shoot method, extract them for subsequent molecular analysis. LCM preserves the exact morphologies of both the captured cells and the surrounding tissue. The Arcturus XT transfers cells from paraffin-embedded and frozen tissue sample stained slides. The entire process can be monitored and documented, and the images stored in an archiving workstation. Research applications include genomics (differential gene profiling, loss of heterozygosity, microsatellite instability, and gene quantification) and proteomics (two-dimensional protein gels, Western blotting, and immuno-quantification of proteins).
Mass Spectrometry
The Thermo Scientific™ Orbitrap Exploris™ 480 Mass Spectrometer is a quadrupole-Orbitrap™ mass spectrometer which provides high-resolution, accurate-mass (HRAM) data, generating incredibly high resolution measurements of up to 480,000 FWHM at m/z 200. Due to its HRAM capabilities, it can be used for low-abundance, high complexity samples at sub-ppb concentrations and /or sub-ppm mass differences. With the option to extend the mass range up to m/z 8000, it facilitates our in-depth characterization needs from intact, top and middle-down through to peptide mapping.
In addition, it is combined with Thermo Scientific™ UltiMate™ 3000 RSLCnano System and HPG-3200RS PUMP, which has a wide flow-pressure footprint with nano-, capillary-, micro-, and regular-flow options.
Microarray
The Research Core Facility is equipped with a state-of-the-art Affymetrix GeneChip System that consists of the following components:
- A GeneChip Hybridization Oven 645 for automated control of hybridization to the GeneChip arrays.
- A GeneChip Fluidics Station 450for automated washing of chips and labeling of hybridized probes. This station can wash and stain four arrays simultaneously.
- A GeneChip Scanner 3000 7G with Autoloader for obtaining high-resolution images of hybridization signals.
- A GeneChip Workstation that controls the operation of the system, data collection and processing of initial raw data.
- A bioinformatics system including Expression Console, Transcriptome Analysis Console, and Ingenuity Pathway Analysis.
microscopy
Leica TCS SP5 Confocal Microscope
This is a very flexible and fast confocal system for fixed or living samples. The system includes a Leica DMI 6000 CS fully automated inverted microscope with motorized stage, condenser, objective, and filter turrets. The microscope is housed in a Ludin full enclosure incubator with an internal Ludin Cube2 with CO2, O2, and humidity controls. It is equipped with 5 lasers for excitation: violet diode (405 nm), multi-line Argon (458, 476, 488, 496, and 514 nm), green HeNe (543 nm), orange HeNe (594 nm), and red HeNe (633 nm); visualization is done using a mercury arc lamp. The spectral beam splitter has freely adjustable bandwidths for the collection of signal in 5 separate detectors simultaneously or sequentially. There is also a transmitted light detector for DIC. There are 9 available objectives, ranging from a 2.5x to a Plan Apo 100x/1.46 NA oil objective. The system runs on the newest version of LAS AF software (LAS AF 2.6.3), with FRAP, FRET, Mark & Find, 3D Visualization, Colocalization, and Live Data Modes.
Nikon A1R Confocal & SIM Super Resolution System
This is a high-speed Nikon A1R confocal microscope combined with a SIM (Structured Illumination Microscopy) super resolution system for fixed and live sample-imaging. The Nikon Eclipse Ti-E inverted microscope is built with fully automated objectives, stage, and filter turrets. The 5 objectives range from 10X to 100X. The confocal components include 5 detectors and 4 lasers: 405nm, 488nm, 561nm, and 640nm in wavelength. The SIM components include a Hamamatsu camera and 3 LED lasers: 488nm, 561nm, and 640nm in wavelength. DAPI images can still be taken under SIM mode. The microscope is capable for bright-field imaging, multichannel fluorescence imaging, large image stitching, multipoint acquisition, time-lapse acquisition, FRAP, FRET imaging, and 2D/3D deconvolution. The resolution for conventional confocal reaches 200-250nm, while the resolution for super resolution system reaches 85-100nm. To allow long hour live cell-imaging, the microscope is housed in an incubator with CO2, O2, and humidity controls.
Nikon Widefield Microscope
The system is built around a Nikon Eclipse TE300 inverted microscope with a range of objectives for phase, DIC, and high resolution epifluorescent imaging. The software package controls a Prior OptiscanTM XYZ stage with a full complement of stage inserts and a Prior filter wheel containing excitation filters from the Chroma 83000 filter set. This set includes single and multiband excitation filters for DAPI, FITC, GFP, Texas Red, Rhodamine, or PI. Fluorescent images are acquired with a black and white ANDOR Neo/Zyla camera; high resolution color images may also be acquired with the DS Fi2 color camera. The stage, filters, shutters, and camera are controlled by the NIS Elements software from Nikon.
Olympus iXplore CSU W1 Spinning Disk
Description coming soon!
Zeiss AxioObserver/Apotome Microscope
The system is built around a Zeiss AxioObserver Z1 inverted fluorescent microscope, fully automated, with component recognition to minimize errors. System components include mercury arc lamp excitation, a Zeiss AxioCamMRm CCD camera with 12-bit dynamic range, extended sensitivity in the near infrared, a fully automated XYZ stage, a complement of objectives ranging from 10x to 100x, and five installed filter sets for DAPI, FITC, narrow band GFP, Rhodamine, and Far Red. The Apotome attachment is designed for precise optical sectioning. The Apotome slides easily into the optical path and projects a grid onto the image plane, which is shifted laterally in three defined steps, with an image collected at each step. A software algorithm then removes any out-of-focus signal. The acquisition software is AxioVision v.4.8, including plug-in options for Inside 4D, 3D Deconvolution, Colocalization, Mark & Find, Mosaic, and more.
Zeiss LSM 510 NLO Confocal/Multiphoton Microscope
The Zeiss LSM 510 NLO system is configured to enhance living tissue research. The scanning system is connected to an upright Axioskop 2 FS MOT microscope equipped with a set of objectives selected for physiological measurements and live animal studies. The stage remains in a fixed position, and the objectives have motorized focus control. It is equipped with the following lasers and laser lines for excitation: Argon (458, 477, 488, 514 nm), HeNe (543 nm), HeNe (633 nm), and the Coherent Chameleon-XR Ti: Sapphire laser (tunable from 705 to 980 nm). The ultrafast-pulsed Chameleon laser emitting NIR radiation allows imaging up to 500 μm deep within tissue. There are three PMTs for visible wavelength detection, a transmitted light detector, and two non-descanned detectors for multiphoton imaging. The LSM 510 ZEN operating software includes the Physiology v3.5 and Image Visart v3.5 options that permit 2D, 3D, and 4D image collection and processing, 3D/4D animation, calibration and measurement of ion concentrations, time series analysis, and graphical mean-of-ROI analysis.
Off-line Image Analysis Stations There are two computer stations in the Core Facility Computer Lab reserved for microscope users, which are loaded with specialized imaging software. The RCFoffline2 system (Fujitsu/Siemens Celsius R650) has two processors, four terabyte hard drives, and an ATI Fire GL V7350 video card with 1 GB of on-board memory. It runs on Windows XP Professional (32 bit) and is loaded with Media Cybernetics’ AutoQuant AutoDeblur deconvolution software (AutoQuant X3), including the AutoVisualize option, and full off-line versions of Zeiss LSM 510 AIM (confocal) and Zeiss Axiovision 4.8.2 (AxioObserver with Apotome) software. RCFoffline1, a Dell Optiplex 3010, runs on Windows 7 Professional (64 bit) and is loaded with Leica LAS AF 2.6.3, NIS Elements AR 4.13.04, and Zeiss LSM 510 ZEN 2009.
next generation sequencing
Illumina MiSeq Next Generation Sequencer
The MiSeq is useful for focused applications such as small genome sequencing, metagenomics, amplicon sequencing and targeted gene sequencing. It is capable of generating up to 15 GB of output with 25 M sequencing reads and up to 2 x 300 bp read lengths.
Cluster generation, sequencing and analysis are all performed on the MiSeq. The sequencing process takes place on a single channel flow cell. Multiple samples can be pooled together and sequenced by using unique index barcodes during library preparation
Illumina NextSeq 550 Next Generation Sequencer
The NextSeq 550 is a more flexible system, enabling the analysis of transcriptomes, exomes and whole genomes. It is capable of generating up to 120 Gb of output with up to 400 M single sequencing reads and up to 2 x 150 bp read lengths. The NextSeq 550 also includes the capability of scanning cytogenomic BeadChip arrays.
Real-Time PCR
Bio-Rad CFX96 Fast Touch Real-Time PCR Systems
The Research Core Facility has three Bio-Rad CFX96 Fast Real-Time PCR Systems. The CFX96 is a six-channel real-time PCR system that combines advanced optical technology with precise thermal control to deliver sensitive, reliable detection. The system’s solid-state optical technology (six filtered LEDs, each with a corresponding filtered photodiode) maximizes fluorescence detection for specific dyes in specific channels, providing sensitive detection for quantitation and target discrimination. Data is collected from all wells during data acquisition. At every position and with every scan, the optics shuttle is reproducibly centered above each well, so the light path is always optimal and there is no need to sacrifice data collection on one of the channels to normalize to a passive reference. Users can select multiple data acquisition modes, including a one-color fast scan for SYBR green. Thermal gradient features can be used to optimize reactions in a single run. The new CFX Manager software has advanced analysis tools for performing normalized gene expression. In addition, this system does not require fluorescein or ROX for instrument normalization.
Bio-Rad CFX384 Touch Real-Time PCR System
The Research Core Facility has one Bio-Rad CFX384 Touch Real-Time PCR System. The system's solid-state optical technology (5 filtered LEDs and 5 filtered photodiodes) provide precise quantification and multiplex target discrimination. All other acquisition and analysis parameters are identical to the CFX96 Fast System.
Scientific Advisory Board
The LSU Health Shreveport Research Core Facility currently houses and maintains instrumentation for eight separate technologies. Each instrument is operated and/or supported by a Research Specialist and is overseen by an LSU Health Shreveport Scientific Advisor. The role of the scientific advisor is to provide investigators with assistance in the application of each technology to his or her specific experimental needs. These advisors comprise the Scientific Advisory Board (SAB).
Scientific Advisory Board Members:
Kelly Tatchell, PhD
Board Chairman
Research Core Facility Director
Associate Dean of Graduate Studies
Professor, Biochemistry & Molecular Biology
ktatch@lsuhsc.edu
Nick Goeders, PhD
Faculty Representative
Professor and Head, Pharmacology, Toxicology and Neuroscience
Ngoede@lsuhsc.edu
Chris Kevil, PhD
Scientific Advisor, Mass Spectrometry
Vice Chancellor for Research
Dean, School of Graduate Studies
Professor, Pathology
Ckevil@lsuhsc.edu
Hung wen (Kevin) Lin, PhD
Scientific Advisor, Microscopy
Assistant Professor, Nuerology
Hlin2@lsuhsc.edu
Sumitra Miriyala, PhD
Scientific Advisor, Cellular Metabolism
Assistant Professor, Cellular Biology & Anatomy
smiriy@lsuhsc.edu
Hyung Nam, PhD
Scientific Advisor, Mass Spectrometry
Assistant Professor, Pharmacology & Toxicology
hnam@lsuhsc.edu
Rona Scott, PhD
Scientific Advisor and CMTV Genomics Director
Associate Professor, Microbiology & Immunology
Rscott1@lsuhsc.edu
Matthew Woolard, PhD
Scientific Advisor, Flow Cytometry
Associate Professor, Microbiology & Immunology
mwoola@lsuhsc.edu
Research Core Facility
Research Core Facility
Research Core Facility
Research Core Facility
Research Core Facility
Research Core Facility
CONTACT US
Camille Abshire, MS, MDxT-(AAB), CLS (LSBME)
Research Specialist, Genomics
cabsh2@lsuhsc.edu
(318) 675-4174
David Custis, MS
Research Specialist, Flow Cytometry & Cellular Metabolism
dcusti@lsuhsc.edu
(318) 675-5765
Paula Polk, MS
Assistant Director, Research Core Facility
polk@lsuhsc.edu
(318) 675-4939
Chaowei Shang, PhD
Research Specialist, Microscopy
cshang@lsuhsc.edu
(318) 675-8537
Xinggui Shen, PhD
Director, Mass Spec Core
Assistant Professor, Pathology
xshen@lsuhsc.edu
(318) 675-3371
Ellen Speers, PhD
Research Specialist, Mass Spectrometry
espeer@lsuhsc.edu
(318) 675-8179
Kelly Tatchell, PhD
Director, Research Core Facility
Associate Dean of Graduate Studies
ktatch@lsuhsc.edu
(318) 675-7769