high content imaging | Visikol

Maximizing the Potential of In Vitro Immuno-Oncology Models with High Content Imaging

Carol Tomaszewski — Wed, 29 May 2024 11:42:41 +0000

In vitro immuno-oncology models are used to study the interaction of therapeutics with immune cells in a 2D or 3D assay format. These models are used to assess the effect of compounds and therapeutic antibodies on immune cell infiltration or to screen immune cell populations for use in immunotherapy. The 3D cell culture models are more biologically relevant and allow for proper assessment of immunotherapies designed to promote the immune cell response to tumor cells. Visikol’s in vitro immuno-oncology models and high content imaging technology can address limitations by providing a method for improving in vitro studies used in drug discovery and development. In this blog post, we will explore how Visikol’s in vitro immuno-oncology models can be used for high content imaging, allowing for more accurate and reliable data for drug efficacy and safety testing. We will also discuss how Visikol’s 3D cell culture models are more biologically relevant and how their high content imaging technology can help researchers and pharmaceutical companies maximize the potential of their immuno-oncology models.

Visikol’s in vitro Immuno-Oncology Models

Visikol’s in vitro immuno-oncology models and high content imaging technology can address limitations by providing a method for improving in vitro studies used in drug discovery and development. They offer tissue analysis services for immuno-oncology research, which aims to understand how the immune system interacts with the tumor microenvironment and modulate the immune response to reduce tumor growth. They employ multiplex immunofluorescence to simultaneously interrogate multiple cell populations and use automated image processing algorithms to assess the variety and distribution of immune cells within and around tumor tissue. They also offer High Content Screening services to clients utilizing 3D cell models and traditional cell-based assays. They provide a wide range of validated in vitro screening services, rapid custom assay development, and end-to-end compound screening services.

3D Cell Culture Models

3D cell culture models are a method of growing cells that better replicates the complex characteristics of the in vivo microenvironment, such as diffusion gradients and receptor expression. Traditional 2D cell culture is limited in its ability to replicate these characteristics. The technique in which cells are cultured (2D vs. 3D) can substantially alter the drug’s effect on the cells. 3D cell culture models offer a more natural, tissue-mimicking method of cell growth for drug discovery applications.

Visikol’s expertise in 3D cell culture models and high content imaging technology allows for the assessment of multiple endpoints simultaneously at a cellular resolution. This provides richer datasets than traditional assays, enabling the interrogation and quantitation of cellular response of disease models to treatments, stimuli, or alterations in protein expression. The use of imaging-based endpoints allows for examination of the specific effect of compound treatments on specific sub-populations of cells, as well as providing access to more complex measurements than can be accomplished with traditional assay formats.

Visikol’s in vitro immuno-oncology models and high content imaging technology can provide a method for improving in vitro studies used in drug discovery and development. Their 3D cell culture models are more biologically relevant and allow for a proper assessment of immunotherapies designed to promote the immune cell response to tumor cells. Their high content imaging technology can help researchers and pharmaceutical companies maximize the potential of their immuno-oncology models by providing a method for assessing multiple endpoints simultaneously at a cellular resolution. By utilizing these technologies, researchers can obtain more accurate and reliable data for drug efficacy and safety testing, ultimately leading to the development of more effective cancer treatments.

If you’re interested in learning more, please reach out to a member of our team today!

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Visikol Continues to Expand Imaging Capabilities with an Additional High-Content Confocal Imager

Carol Tomaszewski — Thu, 30 Sep 2021 13:00:37 +0000

Visikol has added an additional ImageXpress® Micro Confocal High-Content Imaging System from Molecular Devices to its robust imaging capabilities and now has three high-content confocal systems, several fluorescent slide scanners, a Bruker MuVi SPIM Light Sheet microscope, and various other imagers. Just over two months since its acquisition by BICO, Visikol continues to grow its laboratory capacity, as well as its team. With another imager, Visikol will be better suited to aid researchers in their drug discovery efforts and is now able to support even larger drug discovery campaigns that leverage advanced high-content imaging.

The ImageXpress Micro Confocal system is a high-content imager capable of switching between widefield and confocal imaging and can capture whole organisms, thick tissues, 2D and 3D models, and cellular or intracellular events. The ImageXpress Micro Confocal system is also capable of taking high-quality images, creating multiple image models, customized image acquisition, and accelerated analysis speed.

“We are excited to add another Molecular Devices imager to our lab. As we continue to grow both our advanced cell culture and advanced imaging services, we rely heavily on equipment like the ImageXpress to increase efficiency and improve the data that we provide our clients,” said CEO Michael Johnson, PhD.

The ImageXpress Micro Confocal system will support Visikol’s end-to-end 3D advanced cell culture services and provide the team with the ability to image both simple 2D monolayer models as well as advanced 3D cell culture models. As a company, Visikol prides itself on its ability to leverage advanced cell culture models to provide its clients with the most relevant in vitro assays to address their research questions. At the heart of these assays is the use of advanced imaging modalities, like those found in the ImageXpress high-content imaging portfolio, to generate enormous quantities of imaging data and turn it into actionable insights using Visikol’s suite of image analysis software.

If you are interested in learning more about Visikol’s imaging capabilities and working on your next high content assay, please click here to contact our team!

Visikol Associate Scientist Kevin Dennis using the ImageXpress® Micro Confocal High-Content Imaging System from from Molecular Devices.

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High Content Screening for Immune Modulators

Griffin Ferrara — Fri, 01 Jan 2021 15:58:42 +0000

Cancer immunotherapies that target T cells via the blockage of immune checkpoints are a promising approach for cancer treatment. However, the response rates vary between cancer patients. Studies have shown that while immunotherapies work well for some cancer patients, other patients fail to respond to the treatment (1). Since T cell activation requires physical contact with antigen presenting cells (APCs) through the formation of immunological synapse (IS) (2), a better understanding of T cell-tumor cell interactions will be critical for the development of next-generation immunotherapy.

When T cells encounter APCs, antigen recognition by CD3/T cell receptor (TCR) complexes trigger a cascade of intracellular signaling that leads to T cell activation. Activated T cells form symmetric interfaces with APCs through the establishment of IS. Kupfer et al. described the IS as a highly organized bull’s eye structure with a central TCR-major histocompatibility complex (MHC) interaction cluster. This cluster is surrounded by LFA-1-ICAM-1 adhesion and CD45 (3, 4). One can visualize the formation of IS through multiplex immunofluorescence (3) to study antigen recognition, adhesion, or co-stimulation/checkpoint receptors between the T cell and the APC. This approach can be adapted to use as a high throughput screening platform for immune modulator of T cells. Numerous studies have shown that immunological synapse can be used to predict effectiveness of chimeric antigen receptor (CAR) cells (5, 6). At Visikol, we believe multiplex high content imaging of IS is an ideal platform for immune modulator drug screening. Therefore, Visikol has established and now offers services for multiplex high content imaging of IS.

As an example, when we co-culture Jurkat cells (T cells) and superantigen-loaded Raji cells (APCs), we can detect IS formation via immunofluorescence (Figure 1) high content confocal imaging and image analysis. Through this approach, we can observe the reorganization of the actin cytoskeleton and adhesion molecules LFA-1 and ICAM-1. At the interface between Jurkat cells (T cells) Raji cells (APCs), there is the formation of a membrane bridge that can be visualized through phalloidin staining. Both phalloidin, LFA-1 and ICAM-1 staining can be detected when Jurkat-Raji cells were in close proximity. Visikol specializes in advanced cell culture assays and imaging and has demonstrated the utility of high content multiplex imaging of IS as a versatile screening platform for immune modulators.

Figure 1: IS formation between Jurkat-Raji cells. IS formed between Jurkat and antigen-loaded Raji cells when they were in close proximity after 45 mins co-culture. Phalloidin (red), ICAM-1 (green), LFA-1 (gray) and DAPI (blue).

References:

Carbone DP et al. First-Line Nivolumab in Stage IV or Recurrent Non-Small-Cell Lung Cancer. N Engl J Med. (2017);376 (25): 2415–2426.
Dustin ML. T-cell activation through immunological synapses and kinapses. Immunol Rev. (2008); 221: 77–89.
Monks, CR et al. Three-dimensional segregation of supramolecular activation clusters in T cells. (1998) 395, 82–86.
Johnson KG et al. A supramolecular basis for CD45 tyrosine phosphatase regulation in sustained T cell activation. Proc Natl Acad Sci U S A (2000); 97:10138–10143.
Xiong et al. Immunological Synapse Predicts Effectivenessof Chimeric Antigen Receptor Cells. Molecular Therapy (2018); 26, 4: 963-975.
Liu et al. The Role of Immunological Synapse in Predicting the Efficacy of Chimeric Antigen Receptor (CAR) Immunotherapy Cell Communication and Signaling (2020); 18:134

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New Visikol HISTO-M Publication: In Vitro High-Content Imaging-Based Phenotypic Analysis of Bronchial 3D Organotypic Air–Liquid Interface Cultures

Mike Johnson — Tue, 11 Aug 2020 01:28:55 +0000

Interested to see how researchers are using tissue clearing to assess lung organoids in their entirety? Researchers from Philip Morris in Switzerland recently published a paper describing their approach to the phenotypic analysis of these models using Visikol HISTO-M and high content fluorescent microscopy.

With the advent of 3D cell culture models, researchers are now able to ask and answer more complex biological questions as well as to model more complex three-dimensional structures such as the lungs. In this publication, the researchers described a novel bronchial 3D organotypic air-liquid interface (ALI) model (hBR-ALI) as well as a unique workflow to image and characterize these models in their entirety. The use of a complex three-dimensional bronchial model presents unique imaging and image analysis problems due to limitations with light penetration and the availability of 3D image analysis software.

To mitigate these problems and to allow for the complete characterization of these models, the researchers leveraged the Visikol HISTO-M tissue clearing approach to reduce overall light refraction which enabled deep three-dimensional imaging.

Throughout the work, the researchers demonstrate the applicability of the technology platform by evaluating two example use cases: Assessing goblet cell hyperplasia in hBR-ALI by using interleukin (IL)-13 and monitoring differentiation status through HCI evaluation markers representative of ciliated epithelial cells and goblet cells (Muc5AC [mucin 5AC]).

The researchers showed that their high content imaging approach was highly applicable for use with imaging models grown on transwell inserts and can be applied to several other models such as skin epithelia and 3D intestinal systems.

If you are interested in leveraging the Visikol HISTO-M technology within your research or learning more about Visikol’s high content imaging and 3D cell culture services, please reach out to our team.

Full Publication

The post New Visikol HISTO-M Publication: In Vitro High-Content Imaging-Based Phenotypic Analysis of Bronchial 3D Organotypic Air–Liquid Interface Cultures first appeared on Visikol.

Visikol Presents on 3D Cell Culture at the NIH

Mike Johnson — Tue, 13 Aug 2019 10:05:28 +0000

The last few years have been very exciting for cell biology and drug discovery in general with the application of iPSC-derived models and the introduction of 3D cell culture models which have great promise for better recapitulating complex in vivo functionalities. From a therapeutic discovery perspective, the hope for these models is that they will better predict in vivo outcomes and thus reduce the time and cost to bring a new therapeutic to market. However, one of the challenges with these complex 3D cell culture models is their characterization in that traditional imaging and analysis approaches can be inadequate and even misleading.

Visikol CEO Dr. Michael Johnson recently gave a talk at the National Institute of Health that was focused on how best to characterize 3D cell culture models. The inherent problem of characterizing 3D cell culture models is similar to regular tissues in that they are too thick and opaque to image through and therefore traditional wide-field or even confocal microscopy only tells you what is going on at the surface of these models. Because the surface cells are most exposed to compounds, nutrients, and oxygen, using this imaging data to extrapolate results can be very misleading as it only tells the story of the cells on the exterior and not the entire cell population. This bias has been detailed by Visikol through a recent application note titled 3D High Content Imaging of Optically-Cleared Microtissues for Screening Antiproliferative Cancer Drugs.

While 3D cell culture models could be embedded and sectioned for H&E and IHC, this process is laborious, low throughput and also greatly reduces an experiments sample size as many 3D cell culture models need to be embedded to acquire a single section from a single model. Additionally, based upon the polarity of a 3D cell culture model, this single slice might not be indicative of the phenotype of interest. Therefore, in this presentation Dr. Johnson goes into great detail on how to use tissue clearing to image the entirety of 3D cell culture models in multiple imaging channels without the need to ever section or remove the models from a well plate. Though this type of processing and confocal microscopy can reduce sample throughput compared to traditional wide-field imaging, it greatly increases the amount of data that can be generated per model, improves the accuracy of the conclusions that are made and also allows for the investigation of complex research questions.

“We see our Visikol® HISTO-M™ workflow as an easy-to-use pipeline for acquiring all of the information from 3D cell culture models but do recognize that for every assay there is a delicate balance between data density (i.e. images per model) and throughput,” described Dr. Johnson. Visikol currently sells the Visikol HISTO-M reagent and also offers end-to-end drug discovery services that leverage this technology combined with Visikol’s expertise in 3D cell culture, imaging and image analysis. Specifically, Visikol offers best-in-class 3D cell culture assays using this platform to address questions such as quantifying immune cell infiltration or assessing antibody therapeutic kinetics. If you are interested in discussing a 3D cell culture project or transitioning your 2D cell culture assay into a 3D cell culture assay reach out today.

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