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Documentation

Confocal microscopy

Learn all about confocal microscopy and what it can do

What is confocal microscopy?

The confocal microscope scans a laser beam through a sample to excite molecules that are in focus. The molecules then emit photons, which are measured by a detector. A pinhole in front of the detector blocks out-of-focus light and hence improves image quality.

Confocal microscopy offers an improved resolution and signal-to-noise ratio over widefield. The ability to vary the height of the focal plane simultaneously while experimenting is also remarkably useful for 3D reconstruction of larger specimens. This method is more time-consuming than widefield in acquisition and image production and generally higher maintenance, in some cases there is also a tradeoff where some resolution is sacrificed to distinguish different structures in a given sample. Confocal Microscopy is an excellent method of optical imaging often preferred by researchers for applications involving visualization of genetic material or structural components of cells. It enables the assessment of the colocalization of structures and has had large ramifications in studies of the viability of cells.
Figure 1: Three color confocal image of Cas9 binding to a single DNA molecule tethered between two optically trapped beads. Blue/green/red markers indicate the position of individual Cas9 complexes.
Solutions

C-Trap

Molecular biology as never seen before

The C-Trap® provides the world’s first dynamic single-molecule microscope to allow simultaneous manipulation and visualization of single-molecule interactions in real time.

Discover the C-Trap

Relevant resources

Learn as much as you can by reading up on our application notes or marathoning our webinars.

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Tuning CAR-T cells by targeting cancer-associated glycan in pancreatic cancer ‍
Tuning CAR-T cells by targeting cancer-associated glycan in pancreatic cancer ‍
Webinar
February 24, 2026
01-01-20

Chimeric antigen receptor (CAR) T cell therapy has transformed cancer treatment, but its efficacy remains limited in solid tumors due to antigen heterogeneity, an immunosuppressive microenvironment, and the glycocalyx barrier. The glycocalyx, composed of dense glycoproteins such as MUC1, is markedly expanded in cancers, where it impedes immune cell access and antigen engagement, thereby reducing therapeutic efficacy. In most adenocarcinomas, the Tn antigen, comprising N-acetylgalactosamine linked to serine or threonine, is overexpressed. Tn-MUC1, a truncated form of MUC1 decorated with Tn antigen, is frequently overexpressed in pancreatic cancer. Here, we incorporate a non-signaling glyco-bridge binder recognizing Tn-MUC1 into mesothelin-directed CAR-T cells. This bridge enhances tumor recognition and cytotoxicity by increasing avidity and facilitating CAR activation in a density- and affinity-dependent manner. To directly validate these effects at the cell interaction level, we used Lumicks z-Movi to quantify CAR-T binding strength to tumor targets. CAR-T cells equipped with the Tn-MUC1 glyco-bridge exhibited higher cell avidity toward Tn-MUC1-expressing tumor cells compared to a CD19 bridge control. To broaden its applicability, we design a tandem Helix pomatia agglutinin (HPA) lectin-based bridge that recognizes Tn antigens across cancer types. CAR-T cells with the HPA-bridge exhibit superior cytotoxicity in pancreatic cancer models.

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Enhancing efficacy against clear cell renal cell carcinoma through format-tuning of bispecific T cell engagers
Enhancing efficacy against clear cell renal cell carcinoma through format-tuning of bispecific T cell engagers
Scientific update
January 29, 2025
01-01-20

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Cell Avidity: a key to accelerate IND filing in cell therapy drug development
Cell Avidity: a key to accelerate IND filing in cell therapy drug development
Whitepaper
July 1, 2023
01-01-20

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Accelerate your cell engager discovery with high throughput measurements of Cell Avidity
Accelerate your cell engager discovery with high throughput measurements of Cell Avidity
Application note
June 1, 2023
01-01-20

T cells play a pivotal role in tumor immunosurveillance. Multispecific cell engagers (CEs) have been adopted in the field of immuno-oncology to redirect T cells toward cancer cells, thereby unleashing the anti-tumor potential of the patient’s immune system. CE-mediated cell binding induces T cell activation and the formation of an immunological synapse, which is a prerequisite for effective tumor cell lysis.

The strength of the initial binding events between a T cell and a tumor cell dictates the efficiency of the anti-tumor response. Assessing cell avidity, i.e. the total intercellular interaction strength between two cells, gives crucial insights into the efficacy of CEs as anti-tumor therapeutic agents.

Here, we deploy LUMICKS’ high throughput avidity measurement (HTAM) technology to measure CE-induced cell avidity in a high throughput manner. We demonstrate the assay performance characteristics, i.e. specificity, precision, and range, via CE titration experiments in the context of a Jurkat T cell model system. We find that the HTAM CA assay is suitable for candidate screening in high throughput, with high sensitivity and precision.

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Cell Therapy Case Study Collection
Cell Therapy Case Study Collection
Brochure
September 8, 2025
01-01-20

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