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Biology subjects

Gillespie, S.

Publications and source records attributed to Gillespie, S..

2 recordsLinked to original sources

Soft, 3D printed muscle ultrasound phantom with structurally tunable B-mode echo intensity

OBJECTIVESImaging phantoms for training and validation are vital to improving the performance and adoption of ultrasound imaging modalities in clinical and pre-clinical applications, and the goal of this study was to assess the viability of 3D printed muscle ultrasound phantoms to meet this need. METHODSWe used a soft stereolithography resin to 3D print phantoms that mimicked the fascicle- and perimysium-scale structure of skeletal muscle and compared the long axis B-mode imaging quality and pattern of the phantom to that of healthy, adult Biceps brachii. We used a pulse-echo, time-of-flight method to measure the acoustic impedance of the resin for comparison to skeletal muscle and common soft tissue mimicking materials. We analyzed the echo intensity (EI) of muscle images to establish a physiological range and compared the EI of different phantom designs to assess the ability to control imaging brightness through structural modification. RESULTSA linear, striated hyper-/hypo-echoic B-mode imaging pattern mimicking long axis Biceps brachii muscle images was achieved with two 3D structure paradigms, rod and honeycomb. Acoustic impedance of Elastic 50A resin is higher than skeletal muscle in bulk, but appears suitable for use in a 3D structured phantom. EI measured in the Biceps images were found to vary both within and across images with an overall mean {+/-} SD of 87 {+/-}13 AU. EI measured in honeycomb phantoms (55 {+/-}15 AU) was higher than in rod phantoms (42 {+/-}13 AU), and a latticed honeycomb further increased EI (90 {+/-}11 AU). CONCLUSIONSThis study serves as proof-of-concept for soft, 3D printed phantoms that replicate the characteristic muscle ultrasound imaging pattern with the ability to tune clinically relevant EI values via structural design.

bioengineering↗

Monomerization of Far-Red Fluorescent Proteins

Anthozoa class red fluorescent proteins (RFPs) are frequently used as biological markers, with far-red emitting variants ({lambda}em ~ 600 - 900 nm) sought for whole animal imaging because biological tissues are permeable to light in this range. A barrier to the use of naturally occurring RFP variants as molecular markers is that all are tetrameric, which is not ideal for cell biological applications. Efforts to engineer monomeric RFPs have usually produced dimmer and blue-shifted variants, as the chromophore is sensitive to small structural perturbations. In fact, despite much effort, only four native RFPs have been successfully monomerized, leaving the vast majority of RFP biodiversity untapped in biomarker development. Here we report the generation of monomeric variants of HcRed and mCardinal, both far-red dimers, and describe a comprehensive methodology for the rapid monomerization of novel red-shifted oligomeric RFPs. Among the resultant variants, is mKelly1 (emission maximum: {lambda}em = 656 nm), which along with the recently reported mGarnet2 (1), forms a new class of bright, monomeric, far-red FPs.

biochemistry↗