The increasing integration between biological and digital interfaces has led to heightened interest in utilizing biological materials to store digital data, with the most promising one involving the storage of data within defined sequences of DNA that are created by de novo DNA synthesis. However, there is a lack of methods that can obviate the need for de novo DNA synthesis, which tends to be costly and inefficient. Here, in this work, we detail a method of capturing 2-dimensional light patterns into DNA, by utilizing optogenetic circuits to record light exposure into DNA, encoding spatial locations with barcoding, and retrieving stored images via high-throughput next-generation sequencing. We demonstrate the encoding of multiple images into DNA, totaling 1152 bits, selective image retrieval, as well as robustness to drying, heat and UV. We also demonstrate successful multiplexing using multiple wavelengths of light, capturing 2 different images simultaneously using red and blue light. This work thus establishes a 'living digital camera', paving the way towards integrating biological systems with digital devices.

We propose a novel and flexible DNA-storage architecture, which divides the storage space into fixed-size units (blocks) that can be independently and efficiently accessed at random for both read and write operations, and further allows efficient sequential access to consecutive data blocks. In contrast to prior work, in our architecture a pair of random-access PCR primers of length 20 does not define a single object, but an independent storage partition, which is internally blocked and managed independently of other partitions. We expose the flexibility and constraints with which the internal address space of each partition can be managed, and incorporate them into our design to provide rich and functional storage semantics, such as block-storage organization, efficient implementation of data updates, and sequential access. To leverage the full power of the prefix-based nature of PCR addressing, we define a methodology for transforming the internal addressing scheme of a partition into an equivalent that is PCR-compatible. This allows us to run PCR with primers that can be variably elongated to include a desired part of the internal address, and thus narrow down the scope of the reaction to retrieve a specific block or a range of blocks within the partition with sufficiently high accuracy. Our wetlab evaluation demonstrates the practicality of the proposed ideas and a 140x reduction in sequencing cost and latency for retrieval of individual blocks within the partition.

ABSTRACT Colony screening in single and multi-species environments is an essential step for microbiome studies. However, it possesses a high possibility of inaccurately classifying the species of interest and demands a high degree of manpower and time. Thus, automating the classification of microbes is beneficial to minimize the time and inaccuracy in the colony screening/picking step. Here, we developed an automated microbial classification system for five target species, based on deep Convolutional Neural Networks (CNN) using images captured by an automated robotic colony picker. Multiple possible scenarios of colony culture and diverse morphologies of colonies were examined in building the training and test datasets to train and validate the model and performance on real-life implementations. The final model trained using 60,000 training images, with 12,000 images per species and 3-fold cross-validation, achieved a test accuracy of 94.2% and a test loss of 0.154. Upon testing using a deployment dataset of 4,500 images (900 images per species) with different methods of applying cells onto the agar plate, high accuracy of up to 96.6% was obtained. Five evaluation metrics were implemented to evaluate diverse scenarios of the test data to justify the validity of the model in real-life applications. This model forms a foundation for the classification of more species through transfer learning in the future.

The field of storing information in DNA has expanded exponentially. Most common modalities involve encoding information from bits into synthesized nucleotides, storage in liquid or dry media, and decoding via sequencing. However, limitations to this paradigm include the cost of DNA synthesis and sequencing, along with low throughput. Further unresolved questions include the appropriate media of storage and the scalability of such approaches for commercial viability. In this review, we examine various storage modalities involving the use of DNA from a systems-level perspective. We compare novel methods that draw inspiration from molecular biology techniques that have been devised to overcome the difficulties posed by standard workflows and conceptualize potential applications that can arise from these advances.

Podosomes are a singular category of integrin-mediated adhesions important in the processes of cell migration, matrix degradation and cancer cell invasion. Despite a wealth of biochemical studies, the effects of mechanical forces on podosome integrity and dynamics are poorly understood. Here, we show that podosomes are highly sensitive to two groups of physical factors. First, we describe the process of podosome disassembly induced by activation of myosin-IIA filament assembly. Next, we find that podosome integrity and dynamics depends upon membrane tension and can be experimentally perturbed by osmotic swelling and deoxycholate treatment. We have also found that podosomes can be disrupted in a reversible manner by single or cyclic radial stretching of the substratum. We show that disruption of podosomes induced by osmotic swelling is independent of myosin-II filaments. The inhibition of the membrane sculpting protein, dynamin-II, but not clathrin, resulted in activation of myosin-IIA filament formation and disruption of podosomes. The effect of dynamin-II inhibition on podosomes was, however, independent of myosin-II filaments. Moreover, formation of organized arrays of podosomes in response to microtopographic cues (the ridges with triangular profile) was not accompanied by reorganization of myosin-II filaments. Thus, mechanical elements such as myosin-II filaments and factors affecting membrane tension/sculpting independently modulate podosome formation and dynamics, underlying a versatile response of these adhesion structures to intracellular and extracellular cues. This article is part of a discussion meeting issue 'Forces in cancer: interdisciplinary approaches in tumour mechanobiology'.