Particle Tracers

Use of natural or synthetic particles as tracers includes at least 4 primary approaches: ‡ Use of particles to visually demonstrate molecular aggregates ‡ Attachment of particles to molecules or cells to mark their locations ‡ Attachment of particles to molecules or cells to allow them to be counted ‡ Attachment of particles to molecules or cells to allow them to be isolated

The aggregates were formed by antigens interacting with polyclonal antisera. flocculation.. e.Molecular Aggregation Many early immunologic assays were based on molecular agglutination.g. the preciptin reaction. clear solutions. or precipitation of molecular aggregates to produce macroscopically visible changes in previously uniform. .plate testing for blood group substances or syphilis. or spot .

containing cells. tannic acid treated red blood cells. or chemically conjugated latex (polystyrene) spheres. .Molecular/Cellular Aggregation Extension of the idea of aggregation to a more quantitative format was provided by microscopic examination of cellular aggregates (rosettes) formed by washed. white blood cells. bacterial cells. Other versions of this used carbon particles. coated with antibodies interacting with antigen molecules or antigen .

Locale was often marked as a color change (reddish with gold colloids) rather than direct visualization of individual particles.150 nm).). As diffusion is inversely related to molecular/particle size.Localization with Particles Microscopic visualization also allowed localization of particle tagged cells or molecules. & even small atomic chelate collections such as decagold. noble metal colloids (5 . . small latex spheres. smaller particles were used in these contexts. however.. large viruses (TMV). e. bacterial cells (coccus sp.g.

lectins.com/books?id=rlcLQmcTADEC&pg=PA1136&lpg =PA1136&dq=noble+metal+colloids&source=bl&ots=yEk7E3Lj_j&sig =Og5QFJqeoOt8o5K3LGeo9QM0tT8&hl=en&ei=rk3USdGGBYbglQf4y sC-DA&sa=X&oi=book_result&ct=result&resnum=3 http://www. or similar markers.pdf Conjugation of molecules to latex or bioparticles involves the same kinds of chemistries already discussed for linking biomolecules to antibodies.in/matersci/bmsjun2000/165.ac. noble metal colloids: http://books.ias.Adsorption of macromolecules to colloids Colloid definition: http://en.google.wikipedia. .org/wiki/Colloid Characterization & modifications.

& sectioning for TEM following etching with alkalis or treatment with microwave heat to unmask antigens. resolution to 10 . can be done using transmission (TEM. embedding.10 nm) or scanning electron microscopy (SEM.50 nm). . They may also be used after fixation & washing for surface structures in SEM. Or after fixation.Localization with Particles (cont. resolution to 1 . including small viruses like lamda phage. Particles or particle conjugates may be introduced into the live organism prior to fixation & embedding for TEM or fixation & carbon/metal coating for SEM.) Localization & counting of individual small particles.

HCG Binding on MA-10 Cells by SEM MA-10 cells in culture Problem: villi & particles of similar size. .

html www.TEM of various particles: www.uk/Chemistry/Staff/brust.devicelink.jp/english/products/html/f_6./microscopy/colloid..co.html .html www.wisc.edu/.ac.com/ivdt/archive/00/03/004.tanaka.ansci.liv..html www.

Automated Particle Counting Counting of particle . . The Fluorescence Activated Cell Sorter (FACS) detects laser . color. or magnetic properties. The Coulter counter evaluates size & number. color. fluorescence. electrical conductivity.tagged molecules or cells may also be done automatically so long as the particles can be detected by size.activated fluorescence (often in attached latex beads). or dimensional profiles. Imaging software tracks contrast.

Chicago. Illinois :1034-1042. Schools of Veterinary Medicine & Engineering.The first Coulter Counter High Speed Automatic Blood Cell Counter and Cell Size Analyzer Wallace H. Coulter Coulter Electronics. Professor of Biomedical Engineering. Purdue University .Paul Robinson. Professor of Immunopharmacology. 1956 J.

com/html_files2/pc2 200.php .cyto.htm http://www.purdue.edu/fl owcyt/educate/pptslide.spectrex.Automated Particle Counting All the FACS & nothing but the FACS: http://www.

edu/cour ses/genomics/method/FACS.davidson.html .edu/COURSES/ Bio111/FACS1.davidson.http://www.bio.bio.gif www.

edu /.emory...Some modern FACS systems such as the BD Aria can examine as many as 9 or even 13 different signals simultaneously./p_pfc.Ex/Em FSC SSC B-530/30 B-576/26 B-610/20 B-695/40 B-780/60 R-660/20 R-710/20 R-780/60 V-450/40 V-530/30 Laser Detector Blue FSC Blue F Blue E Blue D Blue C Blue B Blue A Red C Red B Red A Violet B Violet A .microbiology.5 PE-Cy7 APC Alexa 680 APC-Cy7 Alexa 405 Alexa 430 Laser . www.shtml Dye Sensed FSC SSC FITC PE PE-Texas Red PE-Cy5.

.thermo.html Magnetic particles from Seradyn: http://www.2348 .com/com/cda/landingpage/0.edu/~ols0/intro.lehigh.html .mpikg-golm.de/kc/landfester/ Microscopy of latex & latex films: http://www.Magnetic & latex particles from Bangs Laboratories: http://www.com/learning Formation of mini-emulsions containing carbon or magnetic cores: http://www.bangslabs.00.mpg.

Quantum Dots (Last but not least!) QdotProteinAconjugatewithUV Tunability of Qdot output by adjustment of particle size www.go..com/site /us/en/home/brands/Molecular -Probes/Key-MolecularProbesProducts/Qdot/TechnologyOverview. .aist.htm http://www./FlowIntro.immunology.utoronto.ca/..html ./hot_line/ hot_line_22.invitrogen..html www.jp/.

uk/research/staff/as/research/ .www.aston.ac.ceac.

based on descriptions in Refs 5. the molecules are shown attached to part of the dot for clarity. Quantum dots having different molecules for target-specific interaction.html a./nphoton.150_F1.nature.. c. based on descriptions in ref.2007. 15. Based on the description in ref. The structure of a multimodal QD probe. and. 150 www.Various designs of multimodal QD probes. Zhivko Zhelev.. but extend over the whole surface) and chelators (b.18.19) for nuclear-spin labelling.com /. d. 20. From the article Designing quantum-dot probes Rumiana Bakalova. 487 489 (2007) doi:10.2007 . paramagnetic lipids (a. 3. based on silica-shelled single-QD micelles. . Adapted from ref. attached to the surface.Figure 1 . the silica sphere has QDs and paramagnetic nanoparticles inside and target-specific groups attached to the outside. Ichio Aoki & Iwao Kanno Nature Photonics 1. copyright (2006) ACS.1038/nphoton.b.

HAYWARD. microtubules are green. mitochondria look yellow. a specific protein within the nucleus is pink..nigms.nih.gov/. DNA testing. CA publications. QUANTUM DOT CORP.. Soon the technique may be used for speedy disease diagnosis.. Here./chapter1. or analysis of biological samples. and actin filaments are red.Quantum dot conjugates can simultaneously reveal the fine details of many cell structures.html . the nucleus is blue.

Other New Uses: New uses for particles are modifications of those already mentioned. ‡ Quantum dots are semiconductor aggregates with high optical activity & electron density. gold or silver colloid stains can also be intensified using silver salts. ‡ Uniform latex or gold particles can be used to calibrate electron or visual microscopes. . ‡ Labeling of blots using latex or colloid tagged antibodies or nucleic acids generate colors without added enzyme reactions.

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