Transcript High-Throughput Bioscience Center (HTBC)

High-Throughput Bioscience Center (HTBC)
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Access to rapidly test thousands of
molecules in various bioassays.
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Small Molecule Compound library—114,000
diverse compounds (ChemDiv, Chembridge,
Specs, ChemDiv Kinase)
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Drug Repurposing Libraries—8,770
compounds of known modulators/FDA approved
Drugs (LOPAC, NIHCC, NCI DTP, Micorsource,
Biomol ICCB and FDA)
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Small interfering RNAs (siRNAs)—Whole
genomes targeting 21,000 genes, pools of 4
siRNAs/gene, Human library from Dharmacon,
and Mouse library from Qiagen
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cDNAs—15,000 genes, E. coli stocks Human
ORFeome in Gateway vectors from Open
Biosystems
High-Throughput Assays (384-well plates)
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In vitro targets--Enzymes, Protein-protein
interactions, Ligand-receptor binding, ProteinDNA binding, etc. (10-15,000 compounds/day)
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Fully automated integrated system:
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In vivo cell based-spectrophotometric-Reporter gene-luciferase, proliferation, synthetic
lethality, etc. (15-30,000 compounds per week)
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High-Content Imaging--Mitosis, protein
translocation/location/expression, cellular
morphology, apoptosis, (5-10,000 compounds
per week)
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8, 96 or 384 tip pipettors/pin tools (CaliperLS SciClone
ALH 3000)
Robotic arms/Stackers for moving microplates (Zymark
Twister II)
Robotic 198 plate CO2 Incubator
Integrated plate washer/reagent dispenser (Bio-tek
EL406) and reagent dispenser (Multidrop)
Bar Code reader
Microplate reader (Molecular Devices AnalystGT,
Tecan Infinite M1000)
siRNA Screening at the HTBC
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siARRAY whole Human Genome
siRNA library from ThermoFisher
(formerly Dharmacon)
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80 gene targets per plate
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4 siRNA oligos per gene--pooled
in each well
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267 X 96 well plates targeting
21,122 genes
Library subdivided in:
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10 Kinase (779 genes, can be
tested separately)
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7 GPCR (517 genes)
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76 Druggable genome (6022
genes)
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174 Rest of genome including
XMs (13,804 genes)
All individual siRNA oligos available
for retests of hits
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4 individual oligos per gene
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High-Throughput siRNA assays
performed in 96- or 384-well plates
(384-well preferred)
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Mouse siRNA library from Qiagen
formatted in 384-well plates.
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Agilent system (96/384 well)
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6 stackers
Bravo 96 or 384 tip pipetting head- 9 plate positions
Thermal Plate Sealer
Wellmate Reagent Dispenser (96/384 well)
ELx405 Plate Washer (96/384 well)
High-Content Cellular Imaging
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ImageXpress Micro (Molecular
Devices)
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Robotic Arm Integration
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More Info: http://htbc.stanford.edu
David E. Solow-Cordero, Ph.D.
Director, HTBC, CCSR-0133
[email protected]
3 stackers for fixed plates
Automated incubator for imaging up to 42 livecell plates
Image Analysis Software
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Fully automated inverted
epifluorescence microscope scans 6- to
384-well plates and records multiple
fluorescent images
Live cell option-Temp, CO2, humidity
Xenon Lamp/filter based
Complete spectral range with 15 filter set
combinations
Up to four color imaging/any fluorophore
Phase Contrast and Brightfield
MetaXpress 5 with 10 Analysis modules
2 analysis computers
3rd party/custom/ Matlab, Cell Profiler
Completed fully automated whole
genome siRNA High Content Screen
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200 X 384-well plates imaged, 1 site, 4 colors in
10 weeks; 307,000 images!