Multi-dimensional correlative imaging of subcellular events: combining the strengths of light and electron microscopy
- 245 Downloads
To genuinely understand how complex biological structures function, we must integrate knowledge of their dynamic behavior and of their molecular machinery. The combined use of light or laser microscopy and electron microscopy has become increasingly important to our understanding of the structure and function of cells and tissues at the molecular level. Such a combination of two or more different microscopy techniques, preferably with different spatial- and temporal-resolution limits, is often referred to as ‘correlative microscopy’. Correlative imaging allows researchers to gain additional novel structure–function information, and such information provides a greater degree of confidence about the structures of interest because observations from one method can be compared to those from the other method(s). This is the strength of correlative (or ‘combined’) microscopy, especially when it is combined with combinatorial or non-combinatorial labeling approaches. In this topical review, we provide a brief historical perspective of correlative microscopy and an in-depth overview of correlative sample-preparation and imaging methods presently available, including future perspectives on the trend towards integrative microscopy and microanalysis.
KeywordsCorrelative morphomics Combinatorial labeling Combined microscopy Live-cell imaging Integrated microscopy Electron tomography
The authors acknowledge the facilities, as well as technical and administrative assistance from staff, of the AMMRF at the Australian Centre for Microscopy & Microanalysis of the University of Sydney, and are particularly grateful to Ellie Kable and Deborah Barton. We also thank the Australian Research Council (ARC) for funding some of the research reported herein through ‘Linkage Infrastructure, Equipment and Facilities’- (LE0775598, LE0883030 & LE100100010) and ‘Discovery Project’ grants (DP0881012), as well as support from the ARC/NHMRC FABLS Research Network (RN0460002).
- Agronskaia AV, Valentijn JA, van Driel LF, Schneijdenberg CT, Humbel BM, van Bergen En Henegouwen PM, Verkleij AJ, Koster AJ, Gerritsen HC (2008) Integrated fluorescence and transmission electron microscopy. J Struct Biol 164:183–189Google Scholar
- Albrecht RM, Goodman SL, Simmons SR (1989) Distribution and movement of membrane-associated platelet glycoproteins: use of colloidal gold with correlative video-enhanced light microscopy, low-voltage high-resolution scanning electron microscopy, and high-voltage transmission electron microscopy. Am J Anat 185:149–164CrossRefPubMedGoogle Scholar
- Burton GJ, Thurley KW, Skepper JN (1991) A technique for correlative scanning and transmission electron microscopy of individual human placental villi: an example demonstrating syncytial sprouts in early gestation. Scan Microsc 5:451–458Google Scholar
- Darien BJ, Sims PA, Kruseelliott KT, Homan TS, Cashwell RJ, Cooley AJ, Albrecht RM (1995) Use of colloidal gold and neutron-activation in correlative microscopic labeling and label quantitation. Scan Microsc 9:773–780Google Scholar
- Deerinck TJ, Martone ME, Lev-Ram V, Green DP, Tsien RY, Spector DL, Huang S, Ellisman MH (1994) Fluorescence photooxidation with eosin: a method for high resolution immunolocalization and in situ hybridization detection for light and electron microscopy. J Cell Biol 126:901–910CrossRefPubMedGoogle Scholar
- Donnell C, Hyde B, Dowling EA (1988) Correlative light and scanning electron-microscopy of endometrium in postmenopausal hormone replacement candidates. Mod Pathol 1:A25–A25Google Scholar
- Geissinger HD (1974) A precise stage arrangement for correlative microscopy for specimens mounted on glass slides, stubs or EM grids. J Microsc 100:113–117Google Scholar
- Goodman SL, Albrecht RM (1987) Correlative light and electron-microscopy of platelet-adhesion and fibrinogen receptor expression using colloidal-gold labeling. Scan Microsc 1:727–734Google Scholar
- Jahn K, Barton D, Braet F (2007) Correlative fluorescence- and scanning, transmission electron microscopy for biomolecular investigation. In: Díaz J, Méndez-Vilas A (eds) Modern research and educational topics in microscopy. Formatex Press, Badajoz, pp 203–211Google Scholar
- Nykänen M (2009) Correlation of light with electron microscopy: a correlative microscopy platform. In: Goldys EM (ed) Fluorescence applications in biotechnology and life sciences. Wiley-Blackwell, Hoboken, pp 141–156Google Scholar
- Peachey LD, Ishikawa H, Murakami T (1996) Correlated confocal and intermediate voltage electron microscopy imaging of the same cells using sequential fluorescence labeling, fixation, and critical point dehydration. Scan Microsc 10:237–247Google Scholar