In 2022, Visikol launched a new imaging service in partnership with Standard BioTools Inc, this partnership has allowed us to bolster our imaging capabilities with the support of an industry leader in imaging. With new imaging services, our Business Development team is frequently asked about the use case of each imaging modality. The following few paragraphs will delineate the different imaging modalities and the use cases for each.

The process for designing a study plan requires consideration into one’s desired endpoints. Endpoints can be pathology review, discovery data, or clinical insights. To get to each endpoint, there is a different modality.

Standard Imaging

In the case of standard imaging, H&E, DAB, Masson’s Trichrome (and others) are staples of staining and brightfield imaging. These stains are often easy to use, efficient and cost effective. The use case is for projects of scale, which require simple area defining analysis or pathology review. Brightfield projects also are typically fast turnaround, making it a great fit for a library of samples that need digitization.

Multiplex Immunofluorescent

The next imaging method is IF (immunofluorescent) – specifically multiplexing. Multiplex IF refers to the labeling of a tissue section with more than 2 antibodies and fluorescently imaging antibodies to create an image. These projects are often more labor intense, and expensive than standard imaging. The pricing and turnaround time for these projects are based on complexity. In other words, a sample imaged with 3 antibodies or 20 antibodies will have a major impact on materials needed and effort required to produce images. The use case for IF is for preclinical or clinical tissues with a medium to high throughput sample quantity. IF can prove useful in drug discovery research as well as translational research and is a great starting point for understanding the TME (tumor microenvironment).

Imaging Mass Cytometry (IMC)

Similar to IF imaging, is IMC (Imaging Mass Cytometry). At Visikol, IMC imaging is available through the Hyperion+ Imaging System and provides researchers the opportunity to generate very large amounts of data from small sections of tissue. Unlike IF, IMC allows researchers to choose up to 40 markers per tissue and there is no background noise because the process is a metal-binding labeling approach. While IMC is more expensive than IF, it provides a greater amount of data that is best used for clinical samples in which researchers need to derive as much data as possible out of valuable tissue samples.

At Visikol, we help our clients with all three of the aforementioned imaging approaches. Our team has worked on projects of all types and are happy to help with you about your next project!

In 2021, over 1,000,000 new cases of cancer and over 600,000 cancer deaths are projected to occur in the United States alone. While analysis of processed cell cultures under a microscope can lead to a diagnosis, a pathologist may request a sample of tissue to further determine cancer diagnosis and progression, known as a biopsy.  The understanding of a patient’s biopsy not only aids in the diagnosis and possible treatment routes but can also open the gateway to further understand the progression and the effects of treatment in clinical trial studies. With growing numbers of biopsies per year, more efficient methods of labeling and staining tissues are needed to optimize patient diagnosis, treatment, and clinical trial research. Histological staining and immunofluorescence (IF) for specific biomarkers plays a key role in understanding the patient’s sample fully, however, varied tissue size and incapability to optimize biomarkers and histological staining leads to a loss in vital lifesaving information. This increases the demand for multiplex services which provide labeling with any tumor type, multiple relevant biomarkers, and detailed spatial profiling for a more in-depth understanding of biopsy samples. 

The multiplex fluorescence approach can analyze the cells of a single tissue sample with multiple panels of up to four biomarkers per panel expressed on a single cell level with high precision and accuracy. The basic principle of multiplexing is that multiple protein targets are stained by specific antibodies labeled with distinct fluorophores which are imaged using a fluorescent microscope, stripped of all antibodies, and re-labeled for further analysis. With the limited tissue sample size acquired via biopsy collection such as the punch, surgical, and core needle methods, the multiplex approach vastly opens the door for understanding more about the sample compared to other methods while saving tissue samples for further analysis.  

For example, a biopsy for breast and prostate cancer commonly collected using a core needle method produces a tissue sample the size of a grain of rice. Once collected, the tissue is assessed for diagnosis, stage/classify if cancerous, and estimate prognosis of cancer. These characteristics are determined by tissue tumor markers which are proteins or other substances that are produced by cancer cells compared to normal cells. These markers can also be utilized to indicate whether a patient is a candidate for a particular targeted therapy and assess treatment efficiency. For example, markers such as estrogen receptor and progesterone receptor are tested to determine whether patients with breast cancer should be treated with hormone therapy, or PDL-1, to see if patients with various cancer types should be treated with an immune checkpoint inhibitor. These mentioned markers and more have been characterized and are used in clinical settings, some of which are associated with only one type of cancer, while others can be associated with multiple different cancer types. With the minuscule size of tissue collected from common biopsy procedures, a multiplex technique opens opportunity to obtain the most information from patient biopsies from prognosis, grade, and even possible routes of treatment. 

At Visikol, we’ve launched our own fluorescent multiplex approach which allows our team to examine many biomarkers with clinical value simultaneously on a single formalin-fixed paraffin-embedded (FFPE) tissue section, followed by histological staining to assess changes in tissue architecture on the same tissue sample. This provides extensive information regarding colocalization, correlation and spatial relation between various biomarkers, as well as changes to tissue structure on a single biopsy sample. More importantly, these data points enable high dimensional image processing to generate spatial profiles of a patient-level disease microenvironment. This can ultimately provide more accurate prediction, prognosis, and drug treatment. 

If you are interested in multiplexing biomarkers and how these operations can excel your research, please reach out to our team to discuss your project. We are always interested to work together with our clients as a team to develop customized assays to best suit their needs. 

References

Siegel, RL, Miller, KD, Fuchs, H, Jemal, A. Cancer Statistics, 2021. CA Cancer J Clin. 2021: 71: 7- 33. https://doi.org/10.3322/caac.21654 

Taube JM, Akturk G, Angelo M, Engle EL, Gnjatic S, Greenbaum S, Greenwald NF, Hedvat CV, Hollmann TJ, Juco J, Parra ER, Rebelatto MC, Rimm DL, Rodriguez-Canales J, Schalper KA, Stack EC, Ferreira CS, Korski K, Lako A, Rodig SJ, Schenck E, Steele KE, Surace MJ, Tetzlaff MT, von Loga K, Wistuba II, Bifulco CB; Society for Immunotherapy of Cancer (SITC) Pathology Task Force.. The Society for Immunotherapy of Cancer statement on best practices for multiplex immunohistochemistry (IHC) and immunofluorescence (IF) staining and validation. J Immunother Cancer. 2020 May;8(1):e000155. doi: 10.1136/jitc-2019-000155. Erratum in: J Immunother Cancer. 2020 Jun;8(1): PMID: 32414858; PMCID: PMC7239569.