Corning® Microplates for Microscopy and High Content Imaging

Improve results with microplates for high resolution cell imaging High Performance for Cell-based Assays

Within the drug discovery process, high content screening (HCS) uses sophisticated automated fluorescence microscopy, image acquisition, and analysis to uncover biologically relevant information from cells and compounds. A major challenge for cell-based imaging assays is selecting the ideal combination of plate type, magnification, and image quality that will yield reproducible and robust results. The Corning portfolio provides the highest quality microplates with the broadest range of options to meet this challenge. Cell culture and plating techniques, the use of automation, and plate selection can all help reduce variability and improve results. The use of specialized microplates is especially critical for achieving optimal results. Key plate features include optimized flatness, thickness, film type, well bottom elevation, and surface type as they directly affect the optical quality, cell adherence, and well imaging access.

Enhance Process and Performance Corning has been setting the standard for excellence in life sciences labware for over 85 years. We continue to be an industry leader focused on enhancing your assay process and performance with our cell culture products, microplates, and reagents. All Corning microplates adhere to strict manufacturing standards, process controls, and compliance testing to support consistent, reproducible performance. Corning offers a variety of specialized microplates with a number of clear bottom substrate options to meet all your microscopy and high content imaging needs. They are organized into 3 groups:

◗◗ For 2X - 10X: Polystyrene microplates for standard microscopy and low magnification imaging ◗◗ For 10X - 32X: 384-well and half-area 96-well high content standard base microplates in glass and Cyclic Olefin Copolymer (COC) for increased chemical resistance and high resolution microscopy ◗◗ For 40X and over: 384-well and half-area 96-well high content low bottom elevation microplates in COC for use with high content imagers utilizing high magnification objectives with low numerical aperture immersion lenses All COC and glass 384-well microplates feature a round well geometry that reduces reagent loss due to “wicking” and prevents cells from “clumping” in the corners.

Human osteosarcoma (U2OS) ZFN-engineered stable cell line co-expressing GFP-α-tubulin and RFP-β-actin from MilliporeSigma was imaged at 40X using a Corning 384-well high content low base COC microplate with Molecular Devices ImageXpress® Micro Confocal System.

2 Corning® High Content Screening Microplate Platform Options Cyclic Olefin Copolymer (COC) Standard Low Base (low well Polystyrene Glass Base bottom elevation)

96-well Microplates Not treated n TC-treated n n n Coated n n n

384-well Microplates Not treated n TC-treated n n n Coated n n n

1536-well Microplates Not treated TC-treated n Coated n

High Content Image Analysis Applications

◗◗ High resolution and confocal imaging applications performed at high magnification ◗◗ Molecular localization ◗◗ Signal translocation (temporal changes) ◗◗ Low to high throughput screening applications, including: • Differentiation of primary cells • Assays for apoptosis • Cytotoxicity • Cell migration • Cell proliferation • Neurite outgrowth

3 Corning High Content Screening Solutions for Cell-based Assays

2X - 10X 10X - 32X

Falcon 96-well and 384-well microplates Corning 384-well and half area 96-well standard base high content COC microplates

Polystyrene Microplates for Microscopy Standard Base High Content Cyclic Olefin Copolymer Falcon® black/clear bottom, polystyrene microplates are made (COC) Microplates with crystal-grade virgin polystyrene and are well suited for With an ultra-clear 127 µm thick film bottom and excellent standard fluorescent detection and imaging applications. The whole plate and intra-well flatness, Corning standard base high 190 µm thick microplate bottom enables clear viewing of cells content microplates are ideal for high resolution cellular imaging using low to medium magnification objectives. The microplates applications. The microplates and film are manufactured from are compatible with automation systems and meet standard cyclic olefin copolymer (COC), which has excellent optical proper- ANSI/SBS footprint dimensions. Falcon black/clear bottom ties and mechanical stability as well as chemical resistance, low microplates are offered in standard 96-well and square 384-well auto-fluorescence, and birefringence. formats in both not treated and tissue culture treated surfaces. Corning® BioCoat™ Collagen- and PDL-coated surfaces are also available.

HEK-293 cells grown on Corning CHO-cells grown on Corning Analysis of neurite outgrowth. Representative micrographs of PC-12 cells BioCoat Poly-D-Lysine (PDL) BioCoat Collagen microplates. grown on Corning BioCoat Poly-D-Lysine (PDL)-coated COC high content microplates. Poly-D-Lysine (PDL) Collagen I is an integral extra imaging microplates, after exposure to β-NGF for 7 days. Representative is a synthetic polymer that cellular matrix protein that supports images demonstrate neurite enhances cell adhesion and protein the framework holding cells and outgrowth in the presence of absorption by altering surface tissues together is routinely used 400 ng/mL (left) and 0 ng/mL charges on the culture substrate. in many cell culture applications. (right) β-NGF. Neurites stained Many transfected cells have been Typical examples of cells grown for β-Tubulin, class III (green), and cultured successfully on PDL, as on Collagen I are endothelial cells nuclei stained with Hoechst (blue) well as neuronal cell lines, primary (e.g., HUVEC), hepatocytes, muscle and imaged with a 20X objective neurons, and glial cells. cells, PC12 cells, osteoclasts, or using the Thermo Scientific transfected HEK-293 cells. CellInsight™ Personal Cell Imager).

4 10X - 32X 40X and over

Corning 384-well and half area 96-well high content screening Corning 384-well and half area 96-well high content low base COC glass bottom microplates microplates

High Content Screening (HCS) Glass Bottom High Content Low Base COC Microplates Microplates Corning low base micro- Corning® HCS glass bottom black microplates feature a scratch- plates feature a low well resistant Corning Willow® glass bottom, which help to deliver bottom elevation of 0.3 mm high optical quality. This is ideal for high content cell-based that allows for unobstructed assays performed using imaging systems. The 200 µm glass reading to the edge of the bottom provides a flat and optically clear surface that reduces plate, ensuring you image all autofocus time, increases throughput, and is ideal for imaging the wells, all the time. microscopy. Several pre-coated options are available for optimal With an ultra-clear film cell growth. and excellent flatness, these microplates are ideal for high resolution cellular imaging applications. The microplate and film are manufactured from cyclic olefin copolymer, which has excellent optical properties, chemical resistance, and mechanical stability. The microplates have a well-established tissue culture-treated surface for the attachment and growth of anchorage-dependent cells.

Representative stained HepG2 cells were grown on a Corning BioCoat™ high content screening microplate and were imaged using a 20X objective on a Thermo Scientific CellInsight™ Personal Cell Imager.

Images provided by Ludwig Institute for Cancer Research, San Diego Branch using the Corning low base COC microplate on Yokogawa CV7000.

5 Specifications and Resources

Microplate Selection Guide for Microscopy and High Resolution Imaging

96-well Full and Half Area Microplates

Falcon® Corning® Corning Magnification 2X - 10X 10X - 32X 40X and over Material of Construction Polystyrene COC standard base Glass COC low base Microplate Bottom Thickness 190 µm 127 µm 200 µm 190 µm Not treated – – Cat. No. 4580 – TC-treated Cat. No. 353219* Cat. No. 4680 – Cat. No. 4517 Corning® BioCoat™ Collagen Cat. No. 356649* – Cat. No. 4582 – Corning BioCoat Fibronectin – – Cat. No. 4584 – Corning BioCoat PDL Cat. No. 356640* – Cat. No. 4586 –

*Standard full area microplate.

384-well Microplates

Falcon Corning Corning Magnification 2X - 10X 10X - 32X 40X and over Material of Construction Polystyrene COC standard base Glass COC low base Microplate Bottom Thickness 190 µm 127 µm 200 µm 190 µm Not treated – – Cat. No. 4581 – TC-treated Cat. No. 353962 Cat. No. 4681 – Cat. No. 4518 Corning BioCoat Collagen Cat. No. 356667 – Cat. No. 4583 – Corning BioCoat Fibronectin – – Cat. No. 4585 – Corning BioCoat PDL Cat. No. 356663 – Cat. No. 4587 –

1536-well Microplates

Falcon Corning Corning Magnification 2X - 10X 10X - 32X 40X and over Material of Construction Polystyrene COC standard base Glass COC low base Microplate Bottom Thickness 190 µm 127 µm 200 µm 190 µm Not treated – Cat. No. 4560 – – TC-treated – Cat. No. 4561 – – Corning CellBIND® – Cat. No. 4563 – – Corning BioCoat PDL – Cat. No. 4564 – –

Custom coatings for all brands and sizes is available upon request.

6 Additional Resources Additional resources can be found on www.corning.com/lifesciences.

◗◗ Instrument and Microplate Considerations to Improve Image Capture and Data Generation During High Content Screens (CLS-AN-081) ◗◗ Corning® Half Area 96-and 384-well Black Glass Bottom Microplates (CLS-AN-216) ◗◗ Corning High Content Imaging COC Microplates Prove to be Effective Tools for High Throughput Data Capture and Analysis (CLS-AN-232) ◗◗ Corning High Content Imaging COC Microplates for High Throughput Data Capture and Analysis (CLS-AN-239) ◗◗ Corning BioCoat™ High Content Imaging Glass Bottom Microplates Provide Enhanced Performance for Cell-based Assays (CLS-AN-244) ◗◗ Corning BioCoat High Content Imaging Glass Bottom Microplates for High Throughput Data Capture and Analysis (CLS-AN-303) ◗◗ Comparison of Ultra-Low Attachment Spheroid Microplates and Hanging Drop Microtissue Formation for High Content Screening (CLS-AN-325)

Additional Corning Solutions for HCS and Drug Discovery Applications

Corning spheroid microplates Corning Matrigel® matrix is a Transwell® permeable supports allows you to generate and natural ECM-based hydrogel that is help produce a cell culture analyze 3D multicellular widely used in models for 2D and environment that more closely spheroids in one microplate. 3D cell culture in vitro, enabling resembles an in vivo state allowing you to improve attachment and cells to carry out activities in a differentiation of many cell types. more natural manner. Design Matrigel matrix can also be used to a variety of experiments using better mimic in vivo environments various pore sizes, membrane for 2D and 3D cell culture types, and coatings. applications.

Visit www.corning.com/lifesciences or contact your local Corning representative to see all solutions for high content screening.

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