Cell and Tissue Processing
Cell sorting using flow cytometry is a specialized type of flow cytometry. It provides a method for sorting a heterogeneous mixture of biological cells into two or more containers, one cell at a time, based upon the specific light scattering and fluorescent characteristics of each cell. Cell sorting can separate live cells based on any scatter, fluorescent signals, or combination of both. We have experience separating live vs. dead cells, cells expressing a specific fluorescent protein, or cells expressing a specific surface marker or set of surface markers. After separation, cells can be cultured and/or used in any downstream assay. We have also successfully created multiple single cell clones expressing different levels of specific proteins from multiple cell lines. We collaborate with a partner laboratory that has multiple cell sorting flow cytometers and years of experience sorting cells in multiple ways.
Covance can dissociate tissues into single-cell suspensions by combining mechanical dissociation with enzymatic degradation of the extracellular matrix, which maintains the structural integrity of tissues. The Miltenyi Biotech Dissociation Kit in conjunction with the gentleMACS™ Octo Dissociator, produces gentle, rapid, and effective generation of single-cell suspensions from primary human or mouse tumor tissue or xenografts. The assay has been optimized for a high yield of tumor cells and tumor infiltrating lymphocytes (TILs), while preserving cell surface epitopes. Dissociated cells can be subsequently cultured, stained for internal/external protein markers, isolated using cell sorting, or used in many other downstream assays. Other than tumor tissues, the flow cytometry experts have many years of experience dissociating many different tissues such as spleens, lymph nodes, lungs, and skin.
Covance can provide frozen protein homogenates containing a battery of protease and phosphatase inhibitors or DNA/RNA homogenates in TRIzol® or RNAlater®. We have a BeadBlaster™ 24 Microtube Homogenizer that completely lyses, grinds, and homogenizes a broad variety of biological samples from soft tissue to bone. Even the most difficult and resistant samples are fully homogenized. The stainless steel tube carrier subjects up to 24 samples to an optimized, high speed, 3D motion producing high energy impacts between the samples and micro-beads to release cellular contents. After homogenization, samples can be centrifuged and the supernatant collected for further processing.
*Denotes services performed at validated partner labs
Other standard techniques are available. Custom assay development is also available to meet our clients’ individual needs.