Quantitative imaging of tissue sections using infrared scanning technology.
Quantification of immunohistochemically (IHC) labelled tissue sections typically yields semi-quantitative results. Visualising infrared (IR) 'tags', with an appropriate scanner, provides an alternative system where the linear nature of the IR fluorophore emittance enables realistic quantitative fluorescence IHC (QFIHC). Importantly, this new technology enables entire tissue sections to be scanned, allowing accurate area and protein abundance measurements to be calculated from rapidly acquired images. Here, some of the potential benefits of using IR-based tissue imaging are examined, and the following are demonstrated. Firstly, image capture and analysis using IR-based scanning technology yields comparable area-based quantification to those obtained from a modern high-resolution digital slide scanner. Secondly, IR-based dual target visualisation and expression-based quantification is rapid and simple. Thirdly, IR-based relative protein abundance QIHC measurements are an accurate reflection of tissue sample protein abundance, as demonstrated by comparison with quantitative fluorescent Western blotting data. In summary, it is proposed that IR-based QFIHC provides an alternative method of rapid whole-tissue section low-resolution imaging for the production of reliable and accurate quantitative data.
Roslin Institute, University of Edinburgh, Edinburgh, UK.
Samantha L
SL
Eaton
Roslin Institute, University of Edinburgh, Edinburgh, UK.|Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, UK.
Elizabeth
E
Cumyn
Roslin Institute, University of Edinburgh, Edinburgh, UK.
Declan
D
King
Roslin Institute, University of Edinburgh, Edinburgh, UK.
Rachel A
RA
Kline
Roslin Institute, University of Edinburgh, Edinburgh, UK.
Sarah M
SM
Carpanini
Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, UK.
Jorge
J
Del-Pozo
Roslin Institute, University of Edinburgh, Edinburgh, UK.
Rona
R
Barron
Roslin Institute, University of Edinburgh, Edinburgh, UK.|Euan MacDonald Centre for Motor Neurone Disease Research, Edinburgh, EH16 4SB, UK.
Thomas M
TM
Wishart
D000818:Animals; D001921:Brain / Q000033:anatomy & histology*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D007259:Infrared Rays; D051379:Mice; D008807:Mice, Inbred BALB C; D008853:Microscopy / Q000379:methods*; D023421:Models, Animal
image analysis; infrared fluorescent tags; morphometric analysis; quantitative immunohistochemistry; tissue section imaging