Photoporation is a gentle technology that uses laser-activated nanoparticles to temporarily permeabilize cell membranes for the delivery of biological molecules. Upon laser irradiation, these nanoparticles generate localized physical forces that create transient membrane pores, allowing a wide variety of cargo, including mRNA, siRNA, DNA, CRISPR-Cas9 RNP complexes for gene knockout (KO), functional proteins, and nanobodies, to enter the cytosol via passive diffusion. This precise mechanism minimizes cellular stress, preserving the high viability and natural functionality of treated cells for critical tasks like high-throughput screening, disease modeling, and large-scale genetic engineering.

The platform is exceptionally versatile, as it is directly compatible with both adherent cell monolayers and suspension cells, overcoming common technical barriers such as the need for stressful cell detachment. This makes the technology ideal for engineering sensitive primary cells as well as immortalized cell lines, ensuring cells remain healthy for advanced research. Its high precision facilitates diverse research goals, from spatial delivery to long-term live imaging using markers delivered directly to the cytosol. Furthermore, the platform’s unique gentleness empowers complex applications, ranging from advanced cell-based therapies and fundamental biomedical research to the development of next-generation drug delivery vehicles.

Photoporation leverages photosensitive nanoparticles to transduce pulsed laser irradiation into localized photothermal effects, facilitating the formation of transient membrane pores with nanoscale precision. This focused physical mechanism serves as a gentle alternative to viral transduction, which exploits natural viral entry mechanisms, and electroporation, which utilizes high-intensity electric pulses that can induce systemic stressors such as reactive oxygen species. By concentrating energy deposition strictly at the nanoparticle-cell interface, the technology effectively preserves the natural viability, phenotype, and proliferative capacity of sensitive primary cells and immortalized lines in their native state.

Yes, photoporation is highly compatible with sensitive and hard-to-transfect cells. Research supported by peer-reviewed papers demonstrates successful results across a variety of challenging cell types, including primary human T cells, NK cells, macrophages, neurons, and various stem cells, while preserving high viability and natural functionality.

By leveraging on a precise mechanism that minimizes cellular stress, the platform provides a substantially higher yield of viable, transfected cells compared to standard benchtop methods. Furthermore, the technology is directly compatible with both adherent and suspension cultures, ensuring cells remain in their natural state and avoiding the need for stressful detachment or trypsinization during the workflow.

Photoporation is directly compatible with adherent cells in their native growth state using standard culture recipients. For adherent monolayers, the technology enables efficient intracellular delivery of DNA, RNA, proteins, oligonucleotides, and synthetic compounds directly in situ without detaching them from their substrate. This avoids the cellular stress associated with trypsinization or mechanical detachment, resulting in a gentler treatment that maintains high cell viability and natural morphology.

Yes, photoporation is a versatile tool for delivering diverse payloads, including Cas9 RNP complexes, mRNA, and plasmid DNA, into “hard-to-transfect” suspension cells such as primary human T cells and NK cells. A major operational benefit is that cells can be photoporated in virtually any optically transparent recipient, from standard flasks to multi-well plates of different sizes, eliminating the need for specialized, proprietary transfection devices or cuvettes. The photoporation mechanism is remarkably gentle, ensuring that engineered populations maintain their full proliferative potential, phenotypic characteristics, and functional potency. This approach achieves an optimal balance between high delivery efficiency and cell survival, providing a versatile and scalable platform for robust ex vivo cell engineering across a wide range of fundamental research and therapeutic discovery applications.

The LumiPore^TM photoporation platform is compatible with a comprehensive range of optically transparent recipients, including glass microscopy slides, chamber slides, Petri dishes, culture flasks, and standard multiwell plates from 6 to 1536 wells. It is engineered to maintain consistent delivery efficiency and high cell viability across these diverse configurations, ensuring that performance remains stable irrespective of the culture scale. This architecture facilitates seamless operational scaling without the requirement for protocol re-optimization, allowing for a direct transition between different experimental formats while maintaining high-throughput capabilities.

Furthermore, a patented thin-layer delivery insert decouples reagent volume from culture surface area, reducing the use of expensive payloads by up to 10–20 fold while preserving efficacy and cell viability.

The LumiPore^TM photoporation system utilizes a Galvano scanner to quickly scan the beam across the culture recipient. The process is characterized by high-precision nanosecond laser pulses, typically ranging from 3 to 7 ns in duration.

Because of this high-speed mechanism the platform can photoporate any well plate format in just few seconds per well, or process an entire plate in approximately 8–10 minutes.

Photoporation enables the efficient intracellular delivery of a wide range of membrane-impermeable payloads across diverse cell types. It is particularly effective for nucleic acids, including siRNA for gene silencing, mRNA for transient expression, and plasmid DNA. A major application is CRISPR-based gene editing via the delivery of Cas9 ribonucleoprotein complexes, enabling efficient gene knockout in primary T cells for CAR-T applications. In addition, the platform supports the delivery of functional proteins and peptides, such as nanobodies for high-resolution imaging, MLKL and caspases for signaling studies, and LFn-FlaA for inflammasome research, as well as diagnostic and cell-tracking agents, including gadolinium-based contrast agents and FITC-dextrans up to 500 kDa.

Intracellular delivery with the LumiPore^TM platform has been successfully demonstrated across a wide range of cell types, including commonly used immortalized cell lines (e.g. HeLa, HEK293, Jurkat, H1299, K562 and THP-1), as well as sensitive primary and stem cell populations. The platform is particularly effective in hard-to-transfect immune cells such as primary human T cells (CD4⁺/CD8⁺ including naive subsets), PBMCs, NK cells, macrophages and Hematopoietic Stem Cells. Successful applications further include Mesenchymal Stem Cells, human Embryonic Stem Cells, iPSCs and iPSC-derived cardiomyocytes, as well as specialized models such as neuronal cells, keratinocytes, insulin-producing cells, lung fibroblasts, glioblastoma cells.

Across all these models, LumiPore^TM preserves cell viability, proliferative capacity, and functional integrity.

Additional data and examples are available in peer-reviewed publications and in the Trince white paper and application note section.

When working with a new cell type, usually we recommend starting with a dedicated optimization step before delivering functional cargo. This optimization step is done by performing a screening experiment at a fixed cell density using FITC-dextran as an inert model cargo. Key parameters, including nanosensitizer type, nanosensitizer concentration, and laser power, are systematically screened, while monitoring intracellular delivery and cell viability. FITC-dextran 10 kDa is suitable to model delivery of small cargos (e.g. siRNA or ASOs), whereas FITC-dextran 150 kDa better represents larger cargos (e.g. mRNA or RNPs). Based on these results, two conditions that provide the best balance between delivery efficiency and cell recovery can be selected. In a separate follow-up experiment, apply these same conditions to test your functional cargo, keeping the cell density identical to that used during screening.

When working with a previously untested intracellular compound it is recommended to first establish suitable photoporation conditions using a well-defined model cargo. Perform a screening experiment at a fixed cell density using FITC-dextran to identify conditions that enable efficient membrane permeabilization while maintaining cell viability. During this step, systematically vary key parameters such as nanosensitizer type, nanosensitizer concentration, and laser power. FITC-dextran 10 kDa is suitable to model the delivery of smaller cargos (e.g. siRNA, ASOs, or small proteins), whereas FITC-dextran 150 kDa is more representative of larger or more complex cargos (e.g. mRNA, plasmid DNA, or RNP complexes). Based on delivery efficiency and cell recovery, select two optimal conditions and apply them in a separate follow-up experiment to your compound of interest, keeping the cell density identical to that used during screening.

Yes. LumiPore^TM is designed to support high-throughput screening (HTS), combining speed, scalability, and efficient reagent use. The platform is fully compatible with high-density plate formats, including 384- and 1536-well plates, while maintaining consistent delivery performance and cell viability across wells.

High processing speed represents a key advantage for HTS workflows. An entire multi-well plate can be processed in approximately 8–10 minutes, while individual wells require only seconds of treatment, allowing large-scale screening campaigns to be conducted efficiently and without throughput limitations. LumiPore^TM also supports seamless scalability, allowing workflows to move directly from small-scale screening formats to larger well plates without protocol re-optimization.

Importantly, cells can be treated directly in their native culture recipients and media, eliminating buffer exchanges or cell detachment steps and further streamlining HTS workflows while maximizing data output per run.

Photoporation using the LumiPore^TM platform shows high and reliable reproducibility across wells and plate formats. The system is engineered for uniform performance in multi-well plates, ensuring consistent delivery efficiency and cell viability from well to well, including in high-density formats. Performance remains stable across different culture scales, allowing results obtained in screening plates to be reliably extrapolated to larger well formats without protocol re-optimization. Highly controlled and uniform laser irradiation ensures that all wells are treated under identical conditions, minimizing technical variability and enabling consistent results across plates.

Yes. Photoporation is a remarkably gentle process, since it induces transient membrane disruptions, allowing cells to stabilize quickly after delivery. This rapid recovery supports short-timeline workflows, with functional readouts, delivery assessments, and imaging commonly performed within a few hours post-photoporation.

Because cells are treated directly in their native culture vessels and medium, additional stress from detachment or handling is avoided, further facilitating immediate downstream analysis. Treated cells retain normal morphology, viability, and proliferative capacity shortly after treatment, making the approach well suited for time-sensitive assays with low technical variability.

The Trince insert is used by first removing the cell culture medium and depositing a small volume of cargo solution directly onto the adherent cell monolayer. The insert is then placed onto this droplet, spreading the liquid into a confined, uniform thin layer, typically ~50–300 µm thick, that fully covers the cells during the delivery step. This thin-layer configuration ensures homogeneous cargo–cell contact and can be combined with delivery methods such as photoporation or simple incubation. After the delivery step, the insert is removed and standard culture conditions are restored.

By confining the cargo to a thin liquid layer rather than a bulk volume, the Trince insert method substantially reduces reagent consumption. Depending on the plate format, delivery volumes can be reduced by approximately 5- to 20-fold compared to conventional bulk delivery. For example, in 96-well plates the volume can be reduced from 50 µL to 10 µL (5-fold reduction), while in larger formats such as 6-well plates, cargo usage can be reduced by up to 18.75-fold, without compromising delivery efficiency or cell viability.

Yes. The workflow is specifically suited for automated and semi-automated screening. Photoporation is performed directly in standard multi-well plates and requires minimal manual handling, enabling straightforward integration into robotic liquid-handling pipelines. Adherent cells do not need to be detached, eliminating transfer steps that typically complicate automation, while compatibility with standard user-defined plates minimizes protocol adaptation. The system is controlled via straightforward software, enabling smooth integration into existing automated or semi-automated setups. Together, these features support efficient, reproducible screening with low operator intervention.

After photoporation, the fate of nanosensitizers is largely determined by their material nature. Inorganic nanosensitizers undergo irreversible physical damage upon laser exposure. A single pulse can cause melting and fragmentation into very small remnants, meaning each nanosensitizer is effective for only one photoporation event. These fragments commonly remain associated with or internalized by the cells following incubation. In contrast, organic nanosensitizers are designed to be biologically degradable. Although laser irradiation may alter their structure, they are subsequently processed within endolysosomal compartments. Thus, depending on their material nature, nanosensitizers either persist transiently as inert fragments or are progressively degraded by the cell.

The onboarding process typically includes a two-day on site training session, during which you will perform two initial experiments to become familiar with the platform and the workflow together with a dedicated Field Application Scientist (FAS). Customer support continues well beyond the initial onboarding phase. During the demo and after purchase, the FAS remains your main point of contact, providing ongoing support, answering technical and scientific questions, and helping refine or adapt protocols to ensure optimal performance for your experiments. In addition, access to a customer support portal is provided, offering updated protocols and a comprehensive overview of tested cell–cargo combinations and a direct communication channel to our scientific team for timely support and guidance.

Support for new users starts with an initial meeting focused on introducing the LumiPore^TM platform and its core principles, helping users understand how the technology works and what it can enable. This is followed by a more in-depth scientific discussion, where users can interact directly with Trince’s Scientist team to evaluate their specific application and discuss experimental needs in detail. After the purchase, a dedicated Field Application Scientist remains closely involved, providing follow-up support, answering questions, and assisting with protocol optimization or adjustments as experiments progress.

Yes. Upon installation, Trince provides a structured, hands-on training programme. This includes a two-day session in which users carry out initial experiments to become familiar with the LumiPore platform and its workflow, under the guidance of a dedicated Field Application Scientist. During this phase, system operation and key functionalities are introduced, and your intended applications are reviewed so that experimental conditions can be fine-tuned, enabling a smooth and confident introduction to the technology.

Yes. Photoporation has been validated in multiple peer-reviewed scientific publications, demonstrating its effectiveness, versatility, and compatibility with a wide range of cell types and applications. These studies support the technology’s use in both fundamental research and more advanced delivery and cell engineering workflows.

Scientific references supporting photoporation are available in the “library” section on Trince’s website. This section includes peer-reviewed journal articles, as well as application notes, white papers and conference posters developed through collaborations with academic and industrial partners or based on extensive in-house testing at Trince.