Optical imaging, which includes confocal microscopy, is routinely used in biomedical research for probing cellular morphology, tissue structure and function. A unique feature of this method is the ability to visualise discrete details in the intact biological system across a broad range of scales in both space and time. We specialise in the capture and quantitative analysis of multi-dimensional images, ranging from the subcellular to whole organism level, with a major focus on applying these methods to studies on important diseases, particularly those involving blood and stem cell biology.
The capture and analysis of multi-dimensional data is a multi-stage process, incorporating:
* Experimental design and sample preparation
* Image acquisition
* Image processing and analysis
We work together with researchers with the aim to apply and optimise optical imaging experiments at each stage of this process, while maintaining an emphasis on training and education.
Australia, Deakin University, BSc (Hons)
Australia, Griffith University, PhD
Potts KS, Sargeant TJ, Markham JF, Shi W, Biben C, Josefsson EC, Whitehead LW, Rogers KL, Liakhovitskaia A, Smyth GK, Kile BT, Medvinsky A, Alexander WS, Hilton DJ, Taoudi S. A lineage of diploid platelet-forming cells precedes polyploid megakaryocyte formation in the mouse embryo. Blood. 2014 Jul 25. pii: blood-2014-02-559468. [Epub ahead of print] PubMed PMID: 25079356.
Kan A, Tan YH, Angrisano F, Hanssen E, Rogers KL, Whitehead L, Mollard VP, Cozijnsen A, Delves MJ, Crawford S, Sinden RE, McFadden GI, Leckie C, Bailey J, Baum J. Quantitative analysis of Plasmodium ookinete motion in three dimensions suggests a critical role for cell shape in the biomechanics of malaria parasite gliding motility. Cell Microbiol. 2014 May;16(5):734-50. doi: 10.1111/cmi.12283. Epub 2014 Mar 28. PubMed PMID: 24612056.
Riglar DT, Rogers KL, Hanssen E, Turnbull L, Bullen HE, Charnaud SC, Przyborski J, Gilson PR, Whitchurch CB, Crabb BS, Baum J, Cowman AF. Spatial association with PTEX complexes defines regions for effector export into Plasmodium falciparum-infected erythrocytes. Nat Commun. 2013;4:1415. doi: 10.1038/ncomms2449. PubMed PMID: 23361006; PubMed Central PMCID: PMC3562467.
Dragavon J, Blazquez S, Rekiki A, Samson C, Theodorou I, Rogers KL, Tournebize R, Shorte SL. In vivo excitation of nanoparticles using luminescent bacteria. Proc Natl Acad Sci U S A. 2012 Jun 5;109(23):8890-5. doi: 10.1073/pnas.1204516109. Epub 2012 May 21. PubMed PMID: 22615349; PubMed Central PMCID: PMC3384150.
Josefsson EC, James C, Henley KJ, Debrincat MA, Rogers KL, Dowling MR, White MJ, Kruse EA, Lane RM, Ellis S, Nurden P, Mason KD, O’Reilly LA, Roberts AW, Metcalf D, Huang DC, Kile BT. Megakaryocytes possess a functional intrinsic apoptosis pathway that must be restrained to survive and produce platelets. J Exp Med. 2011 Sep 26;208(10):2017-31. doi: 10.1084/jem.20110750. Epub 2011 Sep 12. PubMed PMID: 21911424; PubMed Central PMCID: PMC3182050.
Riglar DT, Richard D, Wilson DW, Boyle MJ, Dekiwadia C, Turnbull L, Angrisano F, Marapana DS, Rogers KL, Whitchurch CB, Beeson JG, Cowman AF, Ralph SA, Baum J. Super-resolution dissection of coordinated events during malaria parasite invasion of the human erythrocyte. Cell Host Microbe. 2011 Jan 20;9(1):9-20. doi: 10.1016/j.chom.2010.12.003. PubMed PMID: 21238943.
Roncali E, Savinaud M, Levrey O, Rogers KL, Maitrejean S, Tavitian B. New device for real-time bioluminescence imaging in moving rodents. J Biomed Opt. 2008 Sep-Oct;13(5):054035. doi: 10.1117/1.2976426. PubMed PMID: 19021415.
Rogers KL, Martin JR, Renaud O, Karplus E, Nicola MA, Nguyen M, Picaud S, Shorte SL, Brûlet P. Electron-multiplying charge-coupled detector-based bioluminescence recording of single-cell Ca2+. J Biomed Opt. 2008 May-Jun;13(3):031211. doi: 10.1117/1.2937236. PubMed PMID: 18601535.
Rogers KL, Picaud S, Roncali E, Boisgard R, Colasante C, Stinnakre J, Tavitian B, Brûlet P. Non-invasive in vivo imaging of calcium signaling in mice. PLoS One. 2007 Oct 3;2(10):e974. PubMed PMID: 17912353; PubMed Central PMCID: PMC1991622.
Rogers KL, Stinnakre J, Agulhon C, Jublot D, Shorte SL, Kremer EJ, Brûlet P. Visualization of local Ca2+ dynamics with genetically encoded bioluminescent reporters. Eur J Neurosci. 2005 Feb;21(3):597-610. PubMed PMID: 15733079
The imaging lab is working collaboratively with research teams to apply methods that have emerged in recent years, which improve both 3D and 4D image capture and the reconstruction of large-scale tissue areas or whole mount specimens from serial sections. Project links/resources: Kan A et al. Quantitative analysis of Plasmodium ookinete motion in three dimensions suggests a critical role for cell shape in the biomechanics of malaria parasite gliding motility. Cell Microbiol. 2014 May;16(5):734-50. Team members: Mark Scott and Lachlan Whitehead
This project aims to increase sample throughput and minimise manual operations by applying new software tools that increase automation. These methods promise to overcome some of the major challenges in microscopy, such as the capture of live specimens with high spatio-temporal resolution. Team members: Lachlan Whitehead and Mark Scott
The application of quantitative methods for the segmentation and extraction of features in digital images is critical for understanding the statistical importance of observations made in biological imaging. We work with a number of software packages to improve the automation of image segmentation and the export of measured parameters. Where possible, we streamline data analysis using open source and free software, to give researchers greater independence and to encourage wider collaboration. We are also working on ways to manage and process large data sets (up to 100s of gigabytes in size) to ensure we can avoid bottlenecks commonly associated with new technologies, which have continually increasing rates of data output. Image: https://www.wehi.edu.au/sites/default/files/Art-science-retinal-vascilature.png Image caption: Automated computational analysis image of retinal vasculature. Project resource: Sleebs et al. Inhibition of Plasmepsin V activity demonstrates its essential role in protein export, PfEMP1 display, and survival of malaria parasites. PLoS Biol. 2014 Jul 1;12(7):e1001897. Team member: Lachlan Whitehead