Single-sample image-fusion upsampling of fluorescence lifetime images.

Fluorescence lifetime imaging microscopy (FLIM) provides detailed information about molecular interactions and biological processes. A major bottleneck for FLIM is image resolution at high acquisition speeds due to the engineering and signal-processing limitations of time-resolved imaging technology. Here, we present single-sample image-fusion upsampling, a data-fusion approach to computational FLIM super-resolution that combines measurements from a low-resolution time-resolved detector (that measures photon arrival time) and a high-resolution camera (that measures intensity only). To solve this otherwise ill-posed inverse retrieval problem, we introduce statistically informed priors that encode local and global correlations between the two "single-sample" measurements. This bypasses the risk of out-of-distribution hallucination as in traditional data-driven approaches and delivers enhanced images compared, for example, to standard bilinear interpolation. The general approach laid out by single-sample image-fusion upsampling can be applied to other image super-resolution problems where two different datasets are available.

Single-shot time-folded fluorescence lifetime imaging.

Fluorescence lifetime imaging is an important tool in bioimaging that allows one to detect subtle changes in cell dynamics and their environment. Most time-domain approaches currently involve scanning a single illumination point across the sample, which can make imaging dynamic scenes challenging, while single-shot "rapid lifetime determination" can suffer from large uncertainties when the lifetime is not appropriately sampled. Here, we propose a time-folded fluorescence lifetime imaging microscopy (TFFLIM) approach, whereby a time-folding cavity provides multiple spatially sheared replicas of the lifetime, each shifted temporally with respect to a fixed time gate. This provides a robust, single-shot FLIM approach that we experimentally validate across a broad lifetime range on fluorescent beads and Convallaria samples.

Working at the interface of physics and biology: An early career researcher perspective.

We are a network of Early Career Researchers (ECRs) and a Project Manager who are working on UKRI's "Physics of Life" grants which aim to merge ideas and techniques predominantly used in physics and apply them to biological questions. We have been collaborating since early 2021 to share research, experiences, and provide peer to peer support. Interdisciplinary projects are known for presenting challenges, bringing together disparate subjects and people with not only different knowledge bases, methods, and equipment but also varying ways of working and common languages. This has been the subject of commentary by researchers and funders from a management perspective, and we wanted to add to this discourse, using our experience to share the lessons and challenges we have encountered, from an ECR perspective.

3D Imaging from Multipath Temporal Echoes.

Echo location is a broad approach to imaging and sensing that includes both manmade RADAR, LIDAR, SONAR, and also animal navigation. However, full 3D information based on echo location requires some form of scanning of the scene in order to provide the spatial location of the echo origin-points. Without this spatial information, imaging objects in three-dimensional (3D) is a very challenging task as the inverse retrieval problem is strongly ill-posed. Here, we show that the temporal information encoded in the return echoes that are reflected multiple times within a scene is sufficient to faithfully render an image in 3D. Numerical modeling and an information theoretic perspective prove the concept and provide insight into the role of the multipath information. We experimentally demonstrate the concept by using both radio frequency and acoustic waves for imaging individuals moving in a closed environment.

Fluorescence lifetime imaging with a megapixel SPAD camera and neural network lifetime estimation.

Fluorescence lifetime imaging microscopy (FLIM) is a key technology that provides direct insight into cell metabolism, cell dynamics and protein activity. However, determining the lifetimes of different fluorescent proteins requires the detection of a relatively large number of photons, hence slowing down total acquisition times. Moreover, there are many cases, for example in studies of cell collectives, where wide-field imaging is desired. We report scan-less wide-field FLIM based on a 0.5 MP resolution, time-gated Single Photon Avalanche Diode (SPAD) camera, with acquisition rates up to 1 Hz. Fluorescence lifetime estimation is performed via a pre-trained artificial neural network with 1000-fold improvement in processing times compared to standard least squares fitting techniques. We utilised our system to image HT1080-human fibrosarcoma cell line as well as Convallaria. The results show promise for real-time FLIM and a viable route towards multi-megapixel fluorescence lifetime images, with a proof-of-principle mosaic image shown with 3.6 MP.