Novel Imaging Methods for Biomedical Applications

Novel Imaging methods for biomedical applications

Yuval Garini

Imaging methods were greatly developed in the last ten years. Together with the development of bio-chemical techniques and the better understanding of genetics, new applications and instrumentation can now be used in biology, genetics and medical care.

One of the requirements of modern biomedical applications is the simultaneous detection of many probes. By combining methods to detect multiple colors with high resolution imaging, several methods were developed that allow to detect a large number of probes. These methods include multiple color FISH (M-FISH), Spectral Karyotyping (SKY), Combined Binary Spectral Labeling (COBRA-FISH) and others. These methods are based on fluorescence in situ hybridization (FISH), combinatorial labeling and color or spectral imaging. They became a standard tool in molecular cytogenetics for both research and diagnostics.

In the seminar, the physical basics of imaging, color imaging and spectral imaging will be described, as well as its usage in microscopy. One of the methods, spectral karyotyping (SKY) will be described in more details.

Listed below are papers and reviews that can be most useful for understanding the subjects described in the seminar.

Paper / Subject /
Fundamental issues
J. Reichman, Handbook of optical filters for Fluorescence microscopy.
Available on the website: / Color Filters for fluorescence microscopy
Download from:
http://www.chroma.com/handbook.html
http://www.olympusmicro.com/primer / very good website source on microscopy
Development of multicolor FISH
P. M. Nederlof et al., Cytometry 10, 20-27 (1989) / Demonstration of the potential of multicolor FISH.
P. M. Nederlof et al., Cytometry 11, 126-131 (1990) / First usage of combinatorial labeled FISH probes to distinguish more probes than fluorochromes.
P. M. Nederlof et al., Cytometry 13, 839-845 (1992) / First usage of ratio labeled FISH probes.
T. Ried, A. Baldini, T. C. Rand, D. C. Ward, Proc. Natl Acad. Sci., USA 89, 1388-1392 (1992) / Using 7 different FISH probes by combinatorial labeling (3 fluorochromes).
J. G. Dauwerse et al., Human Molecular Genetics 1, 593-598 (1992) / Using up to 12 FISH probes by ratio labeling.
M. R. Speicher, S. Gwyn Ballard, D. C. Ward, Nature Genetics 12, 368-375 (1996). / First M-FISH paper – Observing all 24 human chromosomes.
E. Schröck et al., Science 273, 494-497 (1996). / First SKY paper – Observing all 24 human chromosomes.
Y. Garini et al., Bioimaging 4, 65-72 (1996) / Technical description of SKY
J. Wiegant et al., Cytogenet Cell Genet 63, 73-6 (1993) / Describing the potential of using direct labeled nucleotides.
D. H. Ledbetter, Hum Mol Genet 1, 297-9 (1992) / The 'colorizing' of cytogenetics: is it ready for prime time?
M. Liyanage et al., Nat Genet 14, 312-5 (1996) / First mouse SKY paper
H. J. Tanke et al., Eur J Hum Genet 7, 2-11 (1999) / COBRA FISH: COmbined Binary RAtio labelling
J. Azofeifa et al., Am. J. Hum. Genet. 66, 1684–1688 (2000) / M-FISH with 7 fluorochromes
Quantitative multicolor analysis
Y. Garini et al, Cytometry 35, 214-226 (1999) / S/N analysis of multicolor FISH methods.
CGH
A. Kallioniemi et al., Science 258, 818-21 (1992) / CGH method
S. du Manoir et al., Cytometry 19, 4-9 (1995) / Quantitative CGH.
J. Piper et al., Cytometry 19, 10-26 (1995) / Computer analysis of CGH
Color banding and other methods
S. Muller, P. C. O'Brien, M. A. Ferguson-Smith, J. Wienberg, Cytometry 33, 445-52 (1998) / Color banding method – RxFISH
C. J. Harrison et al., Cancer Genet Cytogenet 116, 105-10 (2000) / Color banding method – RxFISH
I. Chudoba et al., Cytogenet Cell Genet 84, 156-60 (1999) / High resolution multicolor banding with microdisected probes
O. Henegariu, P. Bray-Ward, D. C. Ward, Nat Biotechnol 18, 345-348 (2000) / An alternative multicolor method with 3 fluorochromes Color changing karyotyping (CCK).
W. C. Chan, S. Nie, Science 281, 2016-8 (1998) / Quantum dot bioconjugates for ultrasensitive nonisotopic detection
M. Bruchez, Jr., M. Moronne, P. Gin, S. Weiss, A. P. Alivisatos, Science 281, 2013-6 (1998) / Semiconductor nanocrystals as fluorescent biological labels

Advanced course - Winnipeg - Mar 2003.doc 3/3 Yuval Garini