Cell Culturing Facility
The 330A HTSF Cell Culture Facility is equipped with a BL-2 laminar flow Hood, 2 CO2 incubators, centrifuges, epifluorescent/phase EVOS inverted mcroscope , water bath, -20C freezer, hemacytometer, Sceptor and storage space (each lab keeps their own stock of supplies). The HTSF has two automated cell plating Combidrop instruments for either 96- or 384-well cell seeding. Typically the hood is used for maintenance and seeding of plates of mammalian cells. Careful sanitation processes are followed to prevent cross-contamination. Other accommodations are requested for any bacterial or yeast work.
Agilent V11 Bravo Automated Liquid Handler
The HTSF offers the Agilent Bravo Automated Liquid Handling Platform because it is one of the most compact yet versatile instruments available to the biotech industry. It is equipped for operations with either a 96- or 384 well head and the platform can be customized for a wide range of assays, providing locations for tips, plates, waste, reagents and wash stations on its nine pipettable deck positions. On the HTSF system there is a separate shaker and an incubation station, as well as grippers within each head for the removal of lids or movement of plates. The 96LT head pipette tips (250ml) are accurate between 2-250 ml. The 384 head pipette tips (either 10, 30, or 70ml) give accurate pipetting from 0.3-70 ml. Common uses are spotting compound plates to numerous daughter plates, reverse transfections, media changes, drug additions, and immunofluorescence assays. The automation method writing software, Vworks, presents an easy to navigate interface to create methods or modify a library of methods already developed on it by previous V11 users. For those developing more sophisticated processes the HTSF V11 is fully integrated with 4 plate stackers to assist with multiple automation steps, a plate washer that is gentle enough for attached cell based assays, and finally a plate sealer to apply an appropriate protective film over the plate after processing.
Beckman-Coulter BioMek NX with Span-8
The HTSF also has a Beckman-Coulter Biomek NX with a Span-8 attachment, which means 8 independently functioning tip positions. This instrument can be used for methods like creating dilution series, plate re-arrays, hit picking, and other uses where not all wells are treated the same. The tips that you order for the Span-8 allow for high precision pipetting of a wide range of volumes.
The software driving the method writing uses Beckman-Coulter’s unique point-and-click programming to create protocols easily understandable to researchers who don’t have an automation programming background. Alternatives such as pipetting techniques, liquid types and deck layouts can be chosen from drop-down lists. Method validation features will check your method and find any logical errors. The Estimated Time to Completion (ETC) feature will provide you with an accurate runtime, enabling you to speed up methods development and efficiently plan your instrument use. Variables, worklists and logicals can be used to vary runs without reprogramming.
BioTek Plate Washers
The ELx405 family of washers has been designed to meet the challenges of various applications, reliability, easy maintenance, yet cost effectiveness. The ELx405 series of washers use BioTek Instruments’ patented manifold design to overcome the difficulties presented by high density microplates and allows virtually the same functionality to be applied to 384-well plates as that is expected with 96-well microplate. The dispense and aspiration manifolds are placed into two physically different parts that are arranged on top of each other. The HTSF has the model (ELx405UCWVS (ELx405 Select CW) that is optimized for loosely adherent cells to allow for washing of HTS plates with minimal disruption to cell attachment . Multiple containers are available to create different buffers for washing or fixation steps.
EVOS® FL Color Imaging System
The EVOS® FL Color Imaging System is a fully-integrated, digital, inverted imaging system for fluorescence, colorimetric, and transmitted-light applications. It is easy to use and delivers high-definition images with convenience.
Perkin-Elmer Envision Multilabel Plate Reader
The EnVision Plate reader 2104 is a rapid, sensitive, and versatile multi-use plate reader that acquires luminescence, and absorbence and bottom or top read of fluorescence. Most common fluorescence excitation and emission filters are available. The model at the HTSF is also capable of doing kinetic experiments including glow, flash and dual luminescence. It also comes with a plate stacker for “walk-away” acquisition of multiple plates.
The flexible EnVision can perform fast kinetic measurements, enzyme assays and numerous other cell-based drug discovery assays. The easy-to-use software aids with acquisition setup and analysis.
Automated Epifluorescence Microscopy ImageXpress Micro by Molecular Devices
The ImageXpress Micro ™ cellular imaging system by Molecular Devices is a fully-integrated hardware and software system for high throughput automated epifluorescent cell-based imaging. The HTSF’s system has environmental control of temperature and 5% CO2 and single-channel pipetting for kinetic experiments. The Screening Facility also has licensed the MetaXpress® and MetaMorph cellular image analysis software for high content analysis, although researchers are free to use whatever analysis pipeline that they prefer using the .tif images. Objective selections include 2x,4x,10x,20x,40x,and 60xoil, and filter sets DAPI, CFP, GFP, FITC, YFP, TRITC, Cy3, TxRed, and Cy5.