Lumito presents results from the successful proof of concept phase

In the field of histopathology, pathologists diagnose patients by assessing imaged tissues based on years of experience of looking at similar images. Even with the pathologist’s trained eye, there is a considerable risk for misdiagnosis [1,2]. In such cases, patients receive unnecessary or suboptimal treatment for their disease. On top of that, the cost to the society is considerable. 

For over half a century, tissue samples have been sectioned and stained with haematoxylin and eosin (H&E) so that pathologists can visualise the morphology of cells. For more than thirty years, immunohistochemistry (IHC), an antibody-based staining technique, has been used to indicate the presence of specific proteins within different tissues, types of cells or cell structures. It is common to detect a protein using a DAB chromogenic stain and combine it with a single counterstain to visualise cell nuclei, normally haematoxylin. Fluorescent IHC techniques generate a more quantitative readout but suffer from photobleaching (the tissue can only be imaged a limited number of times before the fluorescent dye is rendered unusable).

The first histological assay performed by Lumito, using a very simple instrument prototype, is directed to show some of the many benefits of the company’s technology compared to other state of the art techniques in the field:

Lumito’s proof of concept nanocrystal reagent can be applied to immunohistochemistry.

Lumito’s proof of concept staining kit and imaging instrument
– enable multiplexing of luminescence and brightfield imaging
– can separate and detect with fidelity a cell subtype in human tissue

The assay builds on many years of experience and is developed by Lumito’s partner TTP, a leading independent technology and product development company, in collaboration with BioIVT’s PHASEZERO service team, which specializes in IHC studies. It is designed to substantiate, beyond any doubt, abovementioned claims. 

Formalin-fixed paraffin-embedded human colon was sectioned and stained by BioIVT using standard staining equipment (autostainer), then transported to TTP where it was visualised in Lumito’s proof of concept imaging instrument.  

The assay focuses on epithelial cells in the mucosa by employing a commercially available primary antibody that targets the protein family of cytokeratins. Specific binding of primary antibody was detected using a secondary antibody, that, in turn, is linked to the nanocrystals or horseradish peroxidase (HRP) enzyme (followed by DAB chromogen). 

 

Figure 1. Brightfield image with haemoxylin stained nuclei (blue-green) and epithelial cells stained for cytokeratin (DAB brown chromogen). The image quality is limited by the very early instrument prototype.

In Figure 1, tissue stained with pan-cytokeratin antibody and visualised with DAB gives the expected result of brown colour in and around the epithelial cells in the mucosa. This is a high-quality DAB stain, however the impreciseness of the colouring should be noted.

 

Figure 2. Brightfield image of human colon counterstained with haematoxylin. The image quality is limited by the very early instrument prototype.

Figure 2 shows a brightfield image of the tissue stained with haematoxylin and nanocrystals. 

 

Figure 3. Luminescence image of cytokeratin (red/white) detection using nanocrystals. The image quality is limited by the very early instrument prototype.

In Figure 3 is the luminescent image of cytokeratin expression in a tissue stained with haematoxylin and nanocrystals. These data are acquired independently of the cell and tissue morphology in Figure 2 but can easily be merged to provide a guide for the eye (Figure 4). The separation greatly speeds up diagnosis by highlighting areas of interest within the tissue section.

 

Figure 4. Brightfield image of haematoxylin stained nuclei (blue-green) overlaid with luminescence from nanocrystals (cytokeratin; red/white). Please note that the colours used to show the luminescence signal are purely user’s choice. The image quality is limited by the very early instrument prototype.

Figure 4 presents an illustration constructed by overlaying two imaging modalities, luminescence and brightfield, of tissue stained with nanocrystals and haematoxylin. The colours used to indicate the luminescent signal can be freely chosen by the user. 

The conclusion from the study is that the assay fully supports the above statements and that the proof of concept work thereby is done.

[1] Cabillic et al. ALK IHC and FISH discordant results in patients with NSCLC and treatment response: for discussion of the question—to treat or not to treat? ESMO Open. 2018; 3(6): e000419.Published online 2018 Sep 17. doi: 10.1136/esmoopen-2018-000419 

[2] Elmore et al. Diagnostic Concordance Among Pathologists Interpreting Breast Biopsy Specimens. JAMA. 2015;313(11):1122–1132. doi:10.1001/jama.2015.1405

Lumito

Lumito is specialised in research and development of imaging technologies for medical applications. The technology is based on so called UCNP:s (Up Converting Nano Particles) and is aiming to increase image quality in biomedical applications.

Lumito´s IPR covers, with UCNP:s as markers, imaging with high quality of for instance tissue samples. The technology has several possible application areas and Lumito has firstly focused on digital pathology.

Lumito has an agreement regarding development of instrument and staining kits with TTP plc (The Technology Partnership) in Cambridge UK.

Lumito shares LUMITO MTF and warrants LUMITO MTF TO1 are traded at the marketplace NGM Nordic MTF.

TTP 

www.ttp.com 

TTP is an independent technology company where scientists and engineers collaborate to invent, design and develop new products and technologies. With a 30-year history of invention, our multidisciplinary teams are able to deliver across the scope of a project, from research through to ideas, design, engineering and manufacture. 

TTP’s state-of-the-art technical facilities are part of Europe’s largest technology hub in Cambridge, UK. Here, we work across a wide spectrum of industries – including health, telecoms, industrials and consumer – to create breakthrough solutions that bring strong commercial value to clients and the benefits of technology to all.

BioIVT 

BioIVT, formerly BioreclamationIVT, is a leading global provider of research models and value-added research services for drug discovery and development. We specialize in control and disease-state biospecimens including human and animal tissues, cell products, blood, and other biofluids. Our unmatched portfolio of clinical specimens directly supports precision medicine research and the effort to improve patient outcomes by coupling comprehensive clinical data with donor samples. Our PHASEZERO® Research Services team works collaboratively with clients to provide target and biomarker validation, phenotypic assays to characterize novel therapeutics, clinical assay development and in vitro hepatic modeling solutions. And as the premier supplier of ADME-toxicology model systems, including hepatocytes and subcellular fractions, BioIVT enables scientists to better understand the pharmacokinetics and drug metabolism of newly-discovered compounds and their effects on disease processes. By combining our technical expertise, exceptional customer service, and unparalleled access to biological specimens, BioIVT serves the research community as a trusted partner in elevating science. For more information, please visit www.bioivt.com or follow the company on Twitter @BioIVT.