+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Expanding the multicolor capabilities of basic confocal microscopes by employing red and near-infrared quantum dot conjugates

, : Expanding the multicolor capabilities of basic confocal microscopes by employing red and near-infrared quantum dot conjugates. Bmc Biotechnology 9(): 49-49

Confocal microscopy is a widely employed methodology in cellular biology, commonly used for investigating biological organization at the cellular and sub-cellular level. Most basic confocal microscopes are equipped to cleanly discriminate no more than four fluorophores in a given sample, limiting the utility of this method for co-localization, co-expression, and other multi-parameter analyses. In this study, we evaluated the use of red and near-infrared emitting quantum dot staining reagents to expand the multi-parameter capabilities of basic confocal microscopes. We modified a three-laser Zeiss Pascal confocal microscope by the addition of two band-pass filters and one long-pass filter for the detection of three different red to near-infrared quantum dot conjugates. We then performed direct comparisons between organic dye- and quantum dot-labeled detection reagents for the detection of subcellular structures. We found that the quality of staining was generally indistinguishable, although quantum dot reagents do have certain limitations, relative to organic dye conjugates. Using the modified Pascal system, three quantum dot conjugates, two organic dye conjugates, and one fluorescent protein, we demonstrated clean discrimination of six distinct fluorescent labels in a single sample. Our data demonstrate that nearly any basic confocal microscope can be modified by the simple addition of appropriate emission filters, allowing the detection of red and near-infrared quantum dot conjugates. Additionally, quantum dot- and organic dye-based secondary reagents can be successfully combined in complex intracellular staining experiments. Substantial expansion of the multi-parameter capabilities of basic confocal instruments can be achieved with a financial investment that is minimal in comparison to instrument replacement or upgrade with additional lasers.


PMID: 19463154

DOI: 10.1186/1472-6750-9-49

Other references

Rombough, C.J.; Jordan, D Jess; Pearl, C.A., 2003: Rana cascadae (cascade frog). Cannibalism. Herpetological Review e; 34(2): 138

Hardy, T.L.; Holland, D.O., 1952: The separation of DL-threonine from DL-aUothreonine. These amino-acids may be separated by chromatography on filter paper, using water-saturated n-butanol containing 10 per cent. diethylamine by volume as mobile phase and allowing 114 hr. for development.

Machado, Mí; Suzuki, Fábio.Augusto.; Mouco, O.Costa.C.; Hernandes, M.Esteves.; Lemos, M.Angélica.B.T.; Maia, L.Nigro., 2005: Positive troponin T in a chagasic patient with sustained ventricular tachycardia and no obstructive lesions on cine coronary angiography. The assessment and stratification of patients with chest pain in the emergency unit may indicate the appropriate therapy for each patient based on the probability of the presence of acute coronary artery disease and on the risk of its major cardia...

Navas, D., 1972: Medusa buds on eirene elliceana gonads leptomedusae eirenidae. Marine Biology (Berlin) 16(3): 210-213

Szaczawinska, Wanda, 1905: Contribution a l'etude des cytotoxines chez les Invertebres. Archit., Paris: 546-563

Chew, H.C.; Lim, S.H., 2005: Electrocardiographical case. A tale of tall T's. Hyperkalaemia. A 63-year-old woman presented at the emergency department (ED) with a history of increasing lethargy and drowsiness. The electrocardiogram (ECG) showed tall peaked T waves with broadening of the QRS interval, suggestive of hyperkalaemia. This pati...

Anand, C.P.; Blackshear, J.; Yadav, D., 2003: Recurrent giant inflammatory polyposis of the sigmoid colon associated with inflammatory bowel disease. American Journal of Gastroenterology 98(9 Supplement): S153, September

Sreenivasan, V.; Bobier, W.R., 2016: Increased onset of vergence adaptation reduces excessive accommodation during the orthoptic treatment of convergence insufficiency. This research tested the hypothesis that the successful treatment of convergence insufficiency (CI) with vision-training (VT) procedures, leads to an increased capacity of vergence adaptation (VAdapt) allowing a more rapid downward adjustment of t...

Lambert, R.W.; Richard, N.R.; Anderson, J.A., 1996: Cornea1 organ culture. Long-term organ culture of human corneas offers a tool for conducting a variety of biochemical, histological, and wound-healing studies on human tissue in vitro. A requirement of any organ culture system is that cell differentiation and structural...

Matsumoto, H.; Baba, E.; Ishikawa, H., 1972: Bacterial flora in the alimentary canal of dogs. 1. Normal flora in various parts of the intestinal tract. Nihon Juigaku Zasshi. Japanese Journal of Veterinary Science 34(5): 255-261