The ability to associate molecules or molecular functions with particular cellular structures is a key element of biological research. Digital imaging methods are significantly expanding the range of information that can be obtained.
The Weis Center for Research possesses the following instrumentation and technical expertise for biological imaging:
- Conventional light and fluorescence microscopy – microscopes (inverted and upright format) are equipped with digital cameras and software for image processing and analysis; microscopes are linked to a central file server to facilitate image file storage and manipulation.
- Immunohistochemistry – IHC analysis of paraformaldehyde-fixed and paraffin-embedded cells and tissues is carried out; immunohistochemical staining is done manually or using a Ventana automated stained; a variety of antigen retrieval protocols are used.
- Laser scanning confocal microscopy – a Leica TCS SP2 laser scanning confocal microscope with prism spectrometer head is used for high resolution imaging and 3D reconstruction of fixed cells and tissues.
- Spinning disk confocal microscopy – An Olympus IX81 inverted microscope with spinning disk unit and high sensitivity CCD camera is used for fluorescence imaging of live or fixed cells and tissues.
- Structured Illumination Microscopy – a Zeiss Axio Observer.Z1 microscope equipped with ApoTome optical sectioning is used to acquire high resolution images as single planes or Z-stacks that can be combined to create 3D images. Available hardware and software allow time-lapse image capture and image-stitching.
- Electron microscopy – a JOEL JEM-1200EX transmission electron microscope is used to obtain high resolution images from fixed cells and tissues.
- Flow cytometry and cell sorting – A BD FACS ARIA II cell sorter with 3 excitation lasers and a photodiode detector for forward scatter is used for mult-channel flow cytometry of labeled cells and cell sorting.
The ability to analyze nucleic acid molecules in biological samples has become an essential element of biological research. The Weis Center for Research possesses the following instrumentation and expertise for nucleic acid analysis:
- Genotype and quantitative PCR analysis -- Genotype analysis of SNPs and other variants and quantitative real-time PCR analysis is carried out using TaqMan and other assay platforms; there are four Applied Biosystems 7500 Fast Real-Time PCR Systems and one TaqMan Open Array Genotyping System. Roche Light Cyclers are also available for quantitative PCR assays.
- DNA extraction – A QiaSymphony SP DNA Robot from Qiagen is used for automated DNA extraction from blood, saliva, and other biologial samples.
- DNA sequencing – A Life Technologies Personal Genome Machine (PGM) uses the Ion Torrent semiconductor chip technology for DNA sequencing and generates up to 1 Gb of sequence data per run.
The MyCode Project is a system-wide biobanking program to collect and bank biological samples (blood, DNA, and tissue) for broad research use; the samples are linkable to data in Geisinger's electronic medical record system and clincial data warehouse to enable genomic and biomarker studies. Biobank participants are enrolled through Geisinger primary care and specialty clinics.
Approximately 90% of patients invited to participate provide consent. In most cases, research blood samples are obtained at the time of clinical blood draws. As of early 2013, more than 40,000 Geisinger patients had consented to participate in the biobanking program, and more than 100,00 samples had been collected and banked.
The MyCode project was recently expanded to include pediatric participants and consent to link parents and children for research purposes. Biobank samples have been used for genetic and biomarker studies related to cardiovascular disease, obesity and metabolic disease, cancer, and other disorders.
For more information abou the MyCode Biobank contact Samantha Fetterolf (email@example.com).