Equipment
The High-throughput Screening Core (HTSC) occupies 1500 sq. ft. of space on the ground floor of the John Morgan Building. This space provides separate wet laboratory for automation, BSL2 tissue culture, -80oC freezer storage, computing, and office areas.
HTSC Major Equipment
Automated Pipetting
The UPenn HTSC is equipped with two Perkin Elmer Janus with 96/384 Modular Dispensing Tool (MDT) and Verispan 8-tip for automated liquid pipetting. The Janus MDT arm is equipped with a 96 well disposable tip head with capabilities of transferring volumes from 5-200 µl, a 384 well disposable tip head for transferring volumes between 0.5- 25 µl, and a serial dilution tool for preparing dose-responses series in column or row formats. We use 384W slotted PinTools (30nL, 50 nL, or 200nL) to transfer organic small molecules dissolved in DMSO from library plates to assay plates. The MDT workstation is integrated with two plate stack units with capacity for up to 50 microplates each. Serial Dilution Tools (SDT) are used to prepare column or row formatted dilution series of compounds in DMSO to quantitatively assess the activity of compounds. The Verispan 8-tip independent pipetting arm is used to prepare custom formats and is designed for use with disposable tips in 20 µl, 200 µl, and 1 ml sizes.
iQue3 Screener
The iQue3 Screener (Sartorius Corporation) is a flow cytometer with robust automated sampling capabilities compatible with high-throughput screening. It is capable of sampling and analyzing suspension cells, beads, surface released adherent cells, secreted proteins, and mixtures of these sample types, collecting up to ~35,000 events per second. The 3-laser configuration (Violet/Blue/Red) has the potential to record up to 13 channels and the detectors have a dynamic range of 7 decades, which allows for the simultaneous measurement of very low and very high fluorescent emissions in the same sample. The auto sampling capabilities of this instrument enables 96W plates to be analyzed in ~5 minutes and a 384W plate in ~20 minutes and volumes as little as 15ul in 384W plates, reducing reagent usage. The ForeCyt Enterprise software enables offline analysis of acquired data remotely.
Tecan D300e
The UPenn HTSC has 2 Tecan D300e digital dispensers which have the capability to precisely dispense nL-pL volumes of DMSO based reagents to 96/384W assay plates. The D300e software has the flexibility to easily modify compounds dispensed and plate formats required for dose-response analysis and drug combination studies, including the bulk preparation of assay ready plates with compounds for drug sensitivity profiling.
Bulk Reagent Dispensing
The facility has multiple multidrop combi for bulk reagent dispensing. These instruments can be programmed to fill specific columns of assay plates as desired by the user and is capable of dispensing 0.5-2500 µl per well.
Automated Microplate Washers
The core has multiple Biotek ELX405 CW plate washers for automated processing of 96/384 well assay plates.
Multi-Mode Plate Reader
The PerkinElmer EnVision Xcite is a multi-mode plate reader capable of measuring standard detection labels in 96-, 384-, and 1536-well plates, including absorbance, fluorescence intensity, luminescence, ultra-luminescence, time-resolved fluorescence, fluorescence polarization, and AlphaScreen (Amplified Luminescence Proximity Homogeneous Assay).
High-Content Screening Reader
The core is equipped with 3 ImageXpress Micro, an inverted, widefield microscope that utilizes laser-based auto-focus to efficiently image phenotypes of fixed- or live-cells in 96- and 384-well microtiter plates. The IXpress Micro is equipped with a solid-state excitation source, a motorized objective bar with PLAN Apo objectives (4X, 10X, 20X, 40X), interchangeable filter cubes for the flexibility to detect the most common fluorophores (DAPI, FITC, Texas Red, and Cy5), and a 4.6 megapixel scientific CMOS camera for large field-of-view imaging. The MetaXpress® Software, built based on the well-established MetaMorph software, drives the IXpress system to acquire high-resolution images, recognize and segment objects, extract and quantify features, and convert this information into numerical data. Image data is stored in a database on servers maintained by UPENN’s Medicine Academic Computing Services. The core has 10 computers accessible for off-line analysis image data.
FLIPR
The Fluorescence Imaging Plate Reader (FLIPRTetra) Cellular Screening system integrates liquid handling with rapid whole plate imaging of fluorescent and luminescent channels for kinetic cell-based GPCR assays (e.g., calcium, cGMP, cAMP) and ion channel assays (e.g., membrane potential). Our system is equipped with excitation LEDs 335-345 nm, 380-390 nm, 470-495 nm and emission filters 475-535 nm and 515-575 nm.
Quantstudio 5
This qPCR instrument is fitted with a 384W head and used for biochemical Differential Scanning Fluorimetry (Thermal shift) assays.
Tissue Culture
The core is equipped with BSL2 level tissue culture capabilities, including 6 ft. & 4 ft. tissue culture hoods, which can house bulk reagent dispensers (e.g. Multidrop combi) to dispense cells into assay plates; 6 CO2 regulated tissue culture incubators (2 with tri-gas capabilities for culturing cells under hypoxic conditions; 2 benchtop centrifuges fitted with microtiter plate holders, and; a Leica DMIL LED inverted phase-contrast microscope for use while passaging and preparing cells for assays.