Xenolight rediject d luciferin substrate

Human skin tissue was collected and excised as described
above. Explants were incubated at 37 °C with 5% CO₂ in Petri dishes with 10 mL of cDMEM. Medium was replaced daily.
Explants were injected ID using a Micro-Fine Demi 0.3 mL syringe (Becton
Dickinson, UK) with 2 μg of fLuc saRNA complexed with pABOL
or PEI in a volume of 100 μL. After 3 days, skin explants were
inverted and the medium was replaced with 5 mL of cDMEM supplemented
with 100 μL of XenoLight RediJect d-luciferin substrate
(PerkinElmer, UK) and imaged on an IVIS FX Pro (Kodak Co., Rochester,
NY, USA) equipped with Molecular Imaging software version 5.0 (Carestream
Health, USA) for 60 min. Signal from each injection site was quantified
using Molecular Imaging software and expressed as relative light units
(p/s).

All animals were handled in accordance with the UK Home Office Animals
Scientific Procedures Act 1986 and with an internal ethics board and
UK government approved project (P63FE629C) and personal license (IC37CBB8F).
Food and water were supplied ad libitum. Female BALB/c
mice (Charles River, UK) 6–8 weeks of age were placed into
groups (n = 5) and housed in a fully acclimatized
room. In vivo imaging was performed as previously
described.^{43 (link)} Mice were injected either
intramuscularly in both hind legs or intradermally with 5 μg
of fLuc saRNA complexed with either pABOL or PEI in a total volume
of 50 μL. After 7 days, the mice were injected intraperitoneally
(IP) with 100 μL of XenoLight RediJect d-luciferin
substrate (PerkinElmer, UK) and allowed to rest for 10 min. Mice were
then anesthetized using isoflurane and imaged on an In Vivo Imaging System (IVIS) FX Pro (Kodak Co., Rochester, NY, USA) equipped
with Molecular Imaging software version 5.0 (Carestream Health, USA)
for 2 min. Signal from each injection site was quantified using Molecular
Imaging software and expressed as relative light units (p/s).

All animals were handled in accordance with the UK Home Office Animals Scientific Procedures Act 1986 and with an internal ethics board (the Animal Welfare and Ethical Review Body (AWERB)), and UK government approved project license (P63FE629C) and personal license (IC37CBB8F). Food and water were supplied ad libitum. Female BALB/c mice (Charles River, UK) 6–8 weeks of age were placed into groups (n = 5) and housed in a full acclimatized room. In vivo imaging was performed as previously described [5 (link)]. Mice were injected intramuscularly (IM) in both hind leg quadriceps with 5 μg of fLuc saRNA formulations in a total volume of 50 μL. After 7 days, the mice were injected intraperitoneally (IP) with 150 μL of XenoLight RediJect™ D-Luciferin substrate (PerkinElmer, UK) and allowed to rest for 10 min. Mice were then anesthetized using isoflurane and imaged on an In Vivo Imaging System (IVIS) FX Pro™ (Kodak Co., Rochester, NY, USA) equipped with Molecular Imaging software version 5.0 (Carestream Health, USA) for 2 min. The signal from each injection site was quantified using Molecular Imaging software and expressed as total flux (p/s).