At Visikol, we have developed a methodological approach to antigen retrieval and tissue permeabilization for thick tissue immunolabeling, optimized for uniformity and consistency across thick pieces of tissues while preserving histological integrity. We work closely with antibody providers to select high quality antibodies which have been validated for use in immunohistochemistry. We offer single and multiplex immunolabeling services for whole-mount tissues, tissue necropsies/biopsies, thick tissue sections (1-2 mm thick), and microtome/cryotome/vibratome-cut tissue sections.
Validated approach using validated antibodies
As experts in immunolabeling, Visikol has worked with hundreds of antibodies for labeling nearly every type of tissue. Our approach to antigen retrieval and tissue permeabilization has been validated on very large pieces of tissue and whole rodent organs, where we are able to label across several millimeters of tissue. We work closely with providers of antibodies, such as Abcam and ThermoFisher, to select only validated, high quality antibodies for use in our immunolabeling procedures. We only work with antibodies that have been validated for immunohistochemistry and immunocytochemistry, ensuring the best results for our Clients.
Using our multiplex immunolabeling protocols, we can label multiple targets within a tissue simultaneously. Projects we conduct with clients typically utilize 3-4 markers which we can conduct simultaneously with an array of fluorophores spanning the light spectrum. For higher plex labeling (4+ markers), we utilize a proprietary technique to strip the antibodies without damaging tissue, so multiple rounds of labeling can be conducted.
Quantitative analysis of immunolabeled tissues
For thin tissue sections (< 100 microns), we utilize confocal microscopy to ensure the highest quality images are obtained. For thicker tissue sections, whole mount preparations, and intact biopsies/necropsies, we employ tissue clearing with Visikol HISTO. Our Visikol® HISTO™ tissue clearing has been validated to ensure that it preserves tissue morphology and histology, and is a reversible process, so that traditional histopathological evaluation can be conducted after 3D imaging and analysis on selected areas of interest if needed for higher resolution studies.
Using confocal or light sheet imaging, we can rapidly image single and multiplex-immunolabeled tissues. We work closely with our Clients on their imaging projects to ensure that we select the ideal imaging parameters for their research question to maximize throughput and minimize time and cost. We then use our purpose-built 3Screen™ image processing and analytical software to quantitatively assess image sets to tabulate cell counts, quantify the colocalization of biomarkers, analyze spatial distribution of targets, and many other endpoints. By using cell-specific labels, biomarkers can be quantified by cell type, sub-region within tissue, percentage expression within and outside of tumors, to name a few examples.
We find that our Clients derive the most value from our ability to transform large multi-dimensional and multi-channel data sets into key actionable insights within a simple and easy-to-interpret report. Due to the complexity of these studies, we work closely with our Clients at the outset to ensure that our analysis approach will address their specific research question.