Equipment

Automated Labcyte Echo Acoustic Dispenser
Automated Labcyte Echo Acoustic Dispenser

 

 

 

 

 

 

  • Nanoliter-scale reagent dispenser
  • Tipless transfer, resulting in dramatic cost savings
  • Improved information about sample integrity and improved transfer quality through the measurement of DMSO or aqueous buffer levels.
  • Elimination of RNAi or compound loss, cross-contamination, carryover, or leachates through non-contact transfer.
  • Extend RNAi library life through miniaturization of assays.
  • Greatly simplified assay development and dilution processes through the flexibility to transfer any volume of liquid from any well to any other well.
  • Robotic integration with 80 microplate ambient storage units.
  • Robotic integration with Agilent PlateLoc heat sealer, Brooks x-Peel desealer, and Agilent Barcode microplate labeler.

Vala Sciences IC200-KIC Automated High Content Imaging System.

 

 

 

 

 

 

  • 5.54Mp sCMOS camera (100fps, full-frame)
  • Plan APO 4X-60X objectives
  • Rapid autofocus (<200ms)on every field
  • Simple optical path
  • High-resolution positioning (<100nm)
  • Solid-state light engine with 5 discrete bands
  • Filter set (DAPI, FITC, TRITC, Cy5)
  • Environmental control (temperature, CO2, N2)
  • Structured Illumination Microscopy module
  • Integrated with a PreciseFlex 400 sample handler and a Liconic STX44 incubator for live cell experiments.
  • Image analyses include cell counting, live/dead assays, subcellular localization, signal transduction pathway analysis, cell cycle, cell morphological analyses, cell migration

Nanotemper MST

NanoTemper MSFROM Greehey CCRI PILOT award
Recently, the Greehey Children’s Cancer Research Institute acquired the Nanotemper MST Monolith high throughput system which is now located in the RNAi/CRISPR Facility. The Monolith now affords researchers a format for high throughput screening to identify small molecules binding to therapeutically relevant proteins, the characterization of protein-drug interactions as well as protein-protein and protein-ligand interactions.  The Nanotemper uses microscale thermophoresis to measure affinities of interaction between biologically relevant molecules. Microscale thermophoresis is based on the detection of a temperature-induced change in fluorescence of a target as a function of the concentration of a non-fluorescent ligand. The observed change in fluorescence is based on two distinct effects. In a microscopic thermal gradient, the changes in the migration of a protein when it is bound by a ligand can be detected. In addition, the microenvironment of a fluorophore attached to a protein also becomes sensitive to changes induced by small-molecule interaction. These changes in fluorescent upon ligand binding can be used for determining binding affinities. An additional feature of the MST technology is that the measurements can be performed in solution and does not require the immobilization of targets or ligands to solid support. An added attraction is that the MST technology is highly sensitive, requiring small quantities of target and ligand and the assays can be measured rapidly.

Capabilities of the Nanotemper Monolith:

  • Measurements of dissociation constants, affinity, stoichiometry, and binding energetics are rapid
  • MST is equally applicable to a very large size range of molecules. The Monolith instrument can detect interactions between molecules as small as ions to large complexes (e.g. liposomes and ribosomes).
  • MST supports measurement under a broad range of solution conditions where measurements are possible in virtually any buffer, including complex detergent mixtures needed for membrane proteins.
  • Measurements can be performed under close to native, physiological assay conditions since measurements are possible in serum or cell lysates.
  • The method provides reliable diagnostics that make straightforward detection of aggregation and other sample related artifacts possible.
  • The method’s unique capillary system ensures low sample consumption, requiring a < 4 μl sample at nM concentration per data point.
  • Due to fluorescence technology, MST is extremely sensitive and can reliably measure sub-nM binding coefficients.
  • The LED combination (Blue/Red) is suitable for labels most commonly used for labeling macromolecules and suitable for labels commonly used in impure cell extracts.
  • Measures dynamics of the interaction of samples labeled in the blue and red fluorescence range.

Nanotemper Tycho
Nanotemper Tycho

Tycho verifies protein quality by looking at the structural integrity (or foldedness) of a protein. As a thermal ramp is applied, the changes in the intrinsic fluorescence, detected at both 350 nm and 330 nm, from tryptophan and tyrosine residues in the protein are measured. These changes in fluorescence signal indicate transitions in the folding state of a protein. The temperature at which a transition occurs is called the inflection temperature (Ti). The fluorescence is recorded during the thermal run, plotted as a ratio, and used to calculate Ti. Its initial value along with the change in signal over the run (Δ ratio) is also recorded.

  • Identify the quality of any protein in 3 minutes
  • Microliters of sample needed in a label-free way
  • Confirm functionality of protein interaction
  • Determine the right buffer recipe for storage or assay development
  • Check for sample presence, amount, and purity
  • Verify the quality of starting material or similarity between batches

Labcyte Acoustic Echo Dispensing System

Labcyte Echo 525

  • siRNA and miRNA library and small molecule compound plating and reagent dispensing
  • Nanoliter-scale reagent dispensing with volumes ranging from 10nl to 200nl.
  • Tipless transfer, resulting in dramatic cost savings
  • Improved information about sample integrity and improved transfer quality through the measurement of DMSO or aqueous buffer levels.
  • Elimination of RNAi or compound loss, cross-contamination, carryover, or leachates through non-contact transfer.
  • Extend RNAi library life through miniaturization of assays.
  • Greatly simplified assay development and dilution processes through the flexibility to transfer any volume of liquid from any well to any other well.
  • Robotic integration with 80 microplate ambient storage units.
  • Robotic integration with Agilent PlateLoc heat sealer, Brooks x-Peel desealer, and Agilent Barcode microplate labeler.

BMG Labtech Pherastar FS Multimode Plate Reader:

• BMG Labtech Pherastar FS multimode plate reader

 

 

 

 

 

 

 

 

High Throughput Biochemical/Gene Expression/ Cell Viability Screens:

  • Measurements of fluorescence intensity, fluorescence polarization, time-resolved fluorescence, TR-FRET, AlphaScreen, luminescence, UV/Vis Absorbance spectra
  • Two reagent injectors for precise kinetic measurements: Capability for simultaneous injection of reagents and measurement, for highly precise kinetic measurements.
  • Measurement modes: Top and Bottom reading, Endpoint and kinetic measurements, Simultaneous Dual Emission Measurements, Well Scanning.
    A 0.1 mm resolution, automatic Z-height focusing optical stage that precisely focuses light onto the cell monolayer in cell-based assays.
  • Well, scanning that enables high-resolution cell layer detection, generating 3D topology maps for quantitative analyses of defined regions within a well in cell-based assays, significantly reducing inter-well variability.
  • Sophisticated user-friendly analysis software is also available. In addition, this analysis software is freely available from BMG Labtech and thus allows users to analyze data on their own computers.

Tecan Freedom Evo Pipetting Station
Tecan Freedom Evo Pipetting Station

 

 

 

 

 

 

  • Compound plating and reagent dispensing.
  • Volumes ranging from 1.0 µl to 70 µl/well into multiple plates (96- and 384-well microplates
  • Contained within a NuAire HEPA filtered enclosure

Dispensing and Plate Washing Capabilities for Biochemical and Cell-Based Assays

BioTek EL406 Dual Bulk Dispenser/Microplate Washer:

  • Low-volume dispensing and plate washing.
  • Specially designed for gentle washing of microplates containing living cells.
  • Integrated with a BioStack microplate stacker.

• Biotek EL406 dual bulk dispenser/microplate washer

 

 

 

General Lab Facilities for Assay Preparation

Cell culture incubators under normoxic and hypoxic conditions
Cell culture hoods
Microfuges and centrifuges
Liquid nitrogen cryopreservation
Refrigerators and freezers