BRITE Facilities, Resources, and Equipment

The BRITE building features 21,000 square feet of classroom and office space, as well as 31,000 square feet of laboratory space to conduct applied research in areas related to biomanufacturing and biotechnology. In addition, there are common equipment rooms and core laboratories typical of those found in biotech companies.

Liquid Handling Workstations

• Biomek NX (Beckman) 96-head and 384-head liquid handler with a 384-well pintool attachment for nanoliter compound dosing; performs serial dilution, biochemical and cell-based assay assembly, high-content imaging preparation, and high-throughput screening
• Biomek FX (Beckman) 96-head liquid handler with a 384-pintool attachment for compound dosing, biochemical and cell-based assay assembly, high-content imaging preparation, and high-throughput screening; installed in the BSL2 lab
• D300 (HP) Nano- and picoliter digital dispenser for compound dosing, direct dilution and combination drug dosing studies
• NSX-1536 (Nanoscreen) 384-well head liquid handler for rapid biochemical assay assembly and high-throughput screening
• Biomek 3000 (Beckman) single and 8-channel liquid handler for compound cherry picking and serial dilution and some assay assembly (96-well plates) and high-throughput screening (HTS)

Plate Readers

• SpectraMax Plus 384 (Molecular Devices) [4 units] UV and visible absorbance monochromator-based microplate reader
• SpectraMax M5 (Molecular Devices) [2 units] multi-label monochromator and filter-based microplate reader for UV and visible absorbance, fluorescence intensity, time-resolved fluorescence, fluorescence polarization, luminescence, etc.
• PheraStar Plus (BMG LabTechnologies) [2 units] multi-label filter-based microplate reader for UV and visible absorbance, fluorescence intensity, time-resolved fluorescence, fluorescence polarization, luminescence, AlphaScreen, etc, with stackers
• ClarioStar (BMG LabTechnologies) multi-label monochromator and filter-based microplate reader for UV and visible absorbance, fluorescence intensity, time-resolved fluorescence, fluorescence polarization, luminescence, etc, with CO2 and hypoxia chamber
• Victor 3 (Perkin Elmer) [2 units] multi-label filter-based microplate reader for UV and visible absorbance, fluorescence intensity, time-resolved fluorescence, fluorescence polarization, luminescence, etc, with injector and stackers
• Envision (Perkin Elmer) multi-label filter-based dual detector microplate reader for UV and visible absorbance, fluorescence intensity, time-resolved fluorescence, fluorescence polarization, luminescence, etc; a microplate stacker and temperature chamber allow for walk away detection
• FLIPR Tetra (Molecular Devices) high-throughput cellular screening system for GPCR, calcium and ion channel assays. Built-in liquid handler and detection device all in one
• 8200 (Applied Biosystems) Cellular Detection FMAT system for HTS of cell- and bead-based assays.
• Microbeta (Perkin Elmer) microplate luminescence counter

Bulk Dispensing Systems

• Multidrop 384 (Thermo) [10 units] microplate dispensers used for cell plating and bulk dispensing of reagents; especially useful for high-content imaging and high-throughput screening of cell-based assays; with RapidStak (Thermo) plate stacker option
• Microfill (Biotek) microplate dispenser used for cell plating and bulk dispensing of reagents; especially useful for high-content imaging and high-throughput screening of cell-based assays
• Nanoquot (Biotek) non-contact microplate nanodispenser
• Elx405 (Biotek) microplate washer with stacker
• Q Fill 2 (Genetix) microplate dispenser with stacker used for cell plate and bulk dispensing of reagents

Other Core Equipment

• Vcode (Velocity 11) microplate barcode printer and labeler
• ALPs-300 (Abgene) microplate heat sealer
• 3308 (Thermo) Stericult CO2 incubators [2 units] dedicated for HTS lab used for HTS of cell-based assays, hypoxia assays, etc

Data Analysis Software

• Jchem (ChemAxon) compound registration software
• Screenable (Screenable Solutions) data analysis software
• CDD Vault (Collaborative Drug Discovery) software to manage compound structure and biological data information
• Cell Profiler software used for image analysis
• GraphPad Prism

Compound Libraries

• 156,000 small molecule, high diversity library (Asinex and ChemDiv)
• 1,280 FDA-approved Chemical Library® for validation purposes.
• 133 NCI Anticancer Drugs.
• Kinase-specific library

This facility supports high-content screening assays for live-cell imaging with environmental controls for hypoxic and normoxic screening conditions, cell health profiling, apoptosis, cell cycle determination, nuclear translocation, sub-cellular protein localization, and more.

• Cytation 5 (BioTek) live-cell imager with BioSpa 8-vessel temperature and CO2-regulated hotel.
• IncuCyte (Sartorius) live-cell imager, with environmental controls
• Cell Insight NXT (Thermo) high-content screening imager
• Pathway 855 (Becton Dickinson) high-content imaging station
• New: CellInsight CX7 LZR (Thermo) 7-channel, laser-based high-content imager with onstage incubator to control temperature, humidity, and CO2 levels for live cell imaging; unit also has confocal capabilities.

Our facilities support aseptic cell culture preparation for cell-based assays and incubation of cultures in a dedicated suite of incubators, biosafety hoods, refrigerators, freezers, water baths and centrifuges.

• Eclipse TS-100 (Nikon) [2 units] inverted phase-contrast microscopes dedicated for cell culture work
• ViCell (Beckman) [2 units] automated cell counters with trypan blue viability measurements
• DS-Fi2 (Nikon) high-definition 5-megapixel camera attached to one dedicated cell culture microscope
• Xcelligence (Roche) RTCA noninvasive electrical impedance monitoring system to quantify cell proliferation, morphology change, and attachment quality in a label-free, real-time manner
• Z2 (Beckman) particle counter and size analyzer used for cell samples
• AAD-1001S (Amaxa) Nucleofector cell transfection device.
• Hypoxia Chambers (Stemcell Technologies) [2 units] plus controllers for hypoxia experiments.

Our equipment allows for single-cell analysis and sorting using antibodies and dyes and stains for multiparameter phenotypic and functional cell measurements. This allows for the examination of subpopulations of a mixed cell culture as well as primary cells. Cell sorters physically separate cell populations based on selected stain/antibody labeling for further treatments or other analyses.

• CytoFlex (Beckman) flow cytometer equipped with three lasers, seven color detectors, and a plate loader
• FACsAria (Becton Dickinson) flow cytometer (2 lasers) and cell sorter

Our equipment includes items that support high-resolution imaging of biological samples, including bright-field, inverted, phase-contrast, and fluorescent microscopes.

• BZ-X710 (Keyence) inverted phase-contrast, fluorescent microscope outfitted with DAPI, GFP, Texas Red, CFP, Alexa filters; 2x, 4x, 10x, 20x, and 40x objectives; CCD high-resolution digital camera for image capture and analysis
• IX51 (Olympus) inverted phase-contrast, fluorescent microscope outfitted with DAPI, EGFP, Mcherry/Texas Red filters; 4x, 10x, 20x, 40x objectives; 2-megapixel monochrome CCD camera for image capture and analysis
• Eclipse TS-100 (Nikon) [3 units] inverted, phase-contrast microscopes
• Eclipse E200 (Nikon) [6 units] upright bright-field microscopes
• CX31 (Olympus) upright compound microscope
• DP27 (Olympus) 5-megapixel, high-resolution CCD color camera attached to an Eclipse E200 (Nikon) microscope
• Eclipse 80i (Nikon) upright bright-field microscope with six viewing eyepieces
• Eclipse E600FN (Nikon) upright fluorescent phase-contrast microscope specialized for patch-clamp experiments (ionic currents in living cells) with a CCD camera (Nikon).
• SMZ 1000 (Nikon) fluorescent dissecting microscope with a DS-Fi1 (Nikon) 5-megapixel, high-resolution cooled color camera used for research of plant characterizations
• Dissecting microscopes [3 units] for plants, flies, and histology
• Atomic Force Microscope used for characterizing the surface features of a sample.

Our facilities offer upstream bacterial fermentation and mammalian cell culture small-scale (2L–14L) bioreactor equipment and metabolic analysis (pH, glucose, and lactate) as well as downstream cell, antibody, and protein processing equipment (e.g., cell disruption, centrifugation, and filtration). This equipment is used to produce teaching antibodies, engineered proteins/antibodies, insect cells, baculoviruses, and adeno-associated-GFP teaching viral vectors.

• BioFlo 320 (Eppendorf) controller used for mammalian cell culture with single-use bioreactors (5L)
• BioFlo 120 (Eppendorf) (five units) controllers with reusable glass bioreactors (2.2L) for bacterial fermentation, with two of the five units also capable of mammalian cell culture using the single-use bioreactors (5L)
• MetaFLEX (Beckman) used for metabolic analyses (pH, glucose, and lactate)
• Blitz (ForteBio) custom assay system for label-free analysis, protein and antibody quantitation and kinetic experiments
• Slice 20 (Sartorius) filtration systems (four units) used for tangential flow filtration (TFF)
• Multitron (Informs HT) (one unit) shaking incubator for shake flask fermentation
• M-110EH (Microfluidics) microfluidizer/homogenizer for cell membrane disruption and protein extraction
• TSCD-II (Terumo) sterile tubing welder used for creating sterile welds between two pieces of compatible tubing
• Rapid Seal II SE340 (Genesis) tube sealer used for disconnecting thermoplastic tubing in a sterile operation
• Easy-Load II (MasterFlex) peristaltic pumps used for moving fluids during mammalian cell culture
• 3915LT (Thermo) Insect cell incubator with orbital shaker flask platform used for growing insect cells for cell and gene therapy applications
• Solaris 4000 (Thermo) CO2 resistant orbital shaker flask platform used for cell culture and gene therapy applications
• Biostat C-DCU (Sartorius) 30L controller and stainless steel (steam in place “SIP”) bioreactor used for teaching demonstrations

Preparative ultracentrifuges and benchtop cooled and ambient centrifuges are available for use for routine biochemistry, molecular biology, and cell culture work.

• Optima L-90K (Beckman) preparative ultracentrifuge, with variable temperature control and variable titanium rotors; used for separating particles into pellets and clarifying suspensions
• Avanti J-26XP (Beckman) high-speed centrifuge used for separation of fermentation and cellular products
• Allegra X-12R (Beckman) [6 units] refrigerated benchtop centrifuges used for separation processes
• Avanti J-15 (Beckman) high-speed benchtop centrifuge used for separation processes

The following pieces of equipment are available to investigate protein stability and aggregation using advanced techniques such as differential scanning calorimetry, self-interaction chromatography, dynamic light scattering, and circular dichroism.

• Biacore X100 (GE) analyzer for label-free analysis of protein interactions, including protein affinity
• CE (Agilent) capillary electrophoresis used to identify and separate molecules (compounds, proteins, or nucleic acids) based on charge, size, and frictional force in a capillary and microfluidic format.
• AKTA Purifier (Amersham) [1 unit] and AKTA FPLC (GE) [2 units] chromatography workstations
• Goniometer (Ramehart) used to measure the contact angle of protein structures and crystallization
• iTC 200 (Microcal) Isothermal microcalorimeter used for label-free measurement of the binding affinity and thermodynamics of biomolecular interactions to understand function and mechanisms at a molecular level
• VP-DSC (Microcal) differential scanning microcalorimeter used to characterize protein stability
• J-815 (Jasco) circular dichroism spectrometer used to measure changes in protein structure
• nanoAcquity (Waters) uPLC mass spectrometer for UV and visible separation analysis of compounds and proteins
• ProStar (Varian) HPLC used to separate a mixture of compounds and identify, quantify and purify the individual components of the mixture
• SPD-20A (Shimadzu) UV-VIS HPLC Detector
• 1100 (Agilent) LC/MS used to separate the sample components and provide structural identity of the individual components with high molecular specificity and detection sensitivity
• Q-Tof Micro (Waters) mass spectrometer used to identify and characterize proteins or perform structural characterization and specificity of small molecules; perform exact mass measurement, peptide sequencing, etc.

The following equipment is available for work on gene expression, nucleic acid amplification, quantitation, qualification, and related research:

• GeneAtlas 377 (Affymetrix) microarray system used to measure the expression levels of genes
• Lumat 3 LB 9508 (Berthold) Tube luminometer used to measure the luminescence of a selected sample to run gene, enzyme, and intracellular calcium assays and ATP measurements.
• Quat Studio 6 Flex (Applied Biosystems) integrated thermal cycler and real-time PCR machine for 96- or 384-well plates used for Taq-Man assays, digital PCR, genotyping and genomic profiling, and real-time PCR (qPCR).
• iQ5 (BioRad) real-time PCR machine attached to an iQ5 thermal cycler (Biorad) used for qualitative and quantitative analysis of amplified DNA.
• NanoDrop 1000 and NanoDrop 2000 (Thermo) [2 units] absorbance spectrophotometers used for nucleic acid concentration and purity analysis, purified protein analysis, cell density analysis, etc
• QuickDrop (Molecular Devices) [2 units] micro-volume absorbance spectrophotometers used for DNA, RNA, and protein quantification
• Odyssey (Li-Cor) Near-infrared imaging system for DNA and protein gels, blots, and plates
• IS4000R (Kodak) chemiluminescent imaging system used for gels and blots
• GelDoc-It2 (UVP) gel doc imaging system used for chemiluminescence and fluorescence; imaging of films with onboard camera

Chemical analyses are used to confirm compound identity and estimate purity, mass, and structure characterization.

• New: 1260 Infinity II (Agilent) HPLC liquid chromatography system used to separate, identify, and quantify components in a mixture
• 500 MHz NMR (Varian) Nuclear Magnetic Resonance (NMR) used to determine the content, purity, and molecular structure of a sample
• 1200 (Agilent) analytical HPLC [2 units] liquid chromatography systems used to separate, identify, and quantify components in a mixture
• 1200 (Agilent) preparative HPLC liquid chromatography system used to separate, identify, quantify, and collect components in a mixture
• 6130 (Agilent) Quadrupole LC/MS [2 units], which performs the dual tasks of separating the components and then providing structural identity of the individual components with high molecular specificity and detection sensitivity
• 7890A (Agilent) gas chromatography system used to detect the purity of newly synthesized compounds
• 5975A (Agilent) mass spectrometer detector attached to the gas chromatography system used to detect mass of compounds
• CombiFlash (Teledyne Isco) HPLC purification systems [4 units] used to detect the purity of a sample.
• 341 (Perkin Elmer) Polarimeter with UV-transmission, HG and Na spectral lamps for optical characterization of new synthesized compounds
• Explorer (CEM Discover) microwave synthesizer for performing organic and inorganic synthetic chemistries
• PureSolv 400 (PureSolv) 3-solvent dispensing system provides air-free dispensing of ultra-dry and de-oxygenated solvents used for organic chemical synthesis reactions
• Spectrum RX1 (Perkin Elmer) FTIR spectrometer used to identify the functional groups and identification and quality control of raw materials or finished products using IR technology
• Fluromax 4 (Horiba) spectrometer used to measure the fluorescence of solids, liquids, powders and thin films
• LS55 (Perkin Elmer) multi-mode spectrometer used to measures fluorescence, phosphorescence, and chemiluminescence or bioluminescence of a liquid, solid, powder, or thin film sample

The following suite of equipment is used to process tissue samples for tissue and cellular analyses:

• iQ 2000 (Biocare Medical) Kinetic slide stainer used for histology, in situ or immunofluorescent staining
• EG1160 (Leica) paraffin embedding device used to prepare tissues and cells
• SM2000R (Leica) sliding microtome used to section paraffin-embedded fixed or frozen tissue samples for slide preparation
• ASP 300S (Leica) enclosed automated tissue processor
• VT1000S (Leica) vibrating blade microtome for high-quality tissue sectioning
• CM 1850 UV (Leica) cryostat used to prepare frozen sections
• SP 830 Plus (Barnstead) spectrophotometer for use in the visible light range