Training

Rapid dissociation of solid tissue into high-quality, viable single cells or intact nuclei is necessary for a wide range of single-cell genomic applications. In this training, researchers will meet with the CoLab to discuss their tissue dissociation needs. Following protocol consultation, researchers will need to buy the appropriate single-use cartridge from UMGC. For cell isolation, researchers should bring their own enzymes and media. For nuclei isolation, UMGC will supply the necessary nuclei isolation buffer. Additionally, an RNase inhibitor is required for the nuclei isolation workflow; this can either be purchased from UMGC or supplied by the researcher. The training will cover operating the Singulator S200 and preparing, loading, and isolating dissociated cells/nuclei samples. Following isolation, viability and count can be measured either by using the LUNA-FL Automated Fluorescent Cell Counter or manually using a hemocytometer. 

Removal of dead cells and debris from complex cellular samples is necessary for a range of applications, including single-cell workflows. Following an introductory consultation of experimental requirements and general workflow, researchers will schedule a hands-on demonstration of the LeviCell EOS. For training, researchers will need to purchase the single-use cartridge and Levitation agent from UMGC. Researchers will be trained in sample preparation, loading, running the system, and collecting the desired cell fractions. Following isolation, cell count and viability can be assessed using the LUNA-FL Automated Fluorescent Cell Counter or manually with a hemocytometer.

Cell suspension visualization and quantification are critical for determining accurate concentration, viability, and suspension quality before use in downstream single-cell protocols. In this training, researchers will learn to operate the LUNA-FL Automated Fluorescent Cell Counter to measure cell viability and count cells. This training will cover operating the LUNA-FL, preparing and loading samples, and evaluating results. Researchers will supply the cells, and the UMGC will provide the stain and slides. 

Training will provide researchers with the technical skills to independently prepare 3’ or 5’ single-cell transcriptomic experiments using the Chromium Single Cell Controller. To begin, researchers will meet with the CoLab to discuss their experimental goals and receive cell sample requirements. The hands-on training will cover GEM generation, cDNA amplification, and library construction, with a focus on best practices. Researchers will learn to submit the intermediate cDNA and the final libraries for QC to the UMGC and learn how to evaluate the results. 

This training is for generating Visium Spatial Gene Expression libraries for sequencing. At this 1.5-day training, researchers will follow the library preparation protocol under the guidance of a UMGC scientist, using the probe elution from the Visium slides prepared upstream by the UMGC. Before a CoLab training date is set, researchers will meet with Bionet and the UMGC to discuss sample preparation, sectioning, and mounting on slides. Researchers have the option to provide samples already cut on slides or request the service through Bionet. 

Training on the QuantStudio 5 is customized based on the researcher's background with quantitative PCR. Part one (optional) is instructional-based, where the CoLab will provide an overview of qPCR, including PCR basics, general chemistry, intercalating dye vs. probe-based reactions, reverse transcription, and what efficiency means and why it's important. Part two (required) is at the instrument, where the CoLab will demonstrate how to use the QS5 and the software, including data review, how to export the data, QC metrics, and how to troubleshoot. Researchers familiar with qPCR can bring a plate to run at training.